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   2018| April-June  | Volume 14 | Issue 55  
    Online since June 28, 2018

 
 
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ORIGINAL ARTICLES
Development of internet technology TIPHAM (Tool for identity of Powdered Herbals through Analytical Microscopy) for microscopic identification of crude herbal drugs
Dharya Singh, Vidhu Aeri, DB Ananthanarayana
April-June 2018, 14(55):213-226
DOI:10.4103/pm.pm_563_17  
Background: Industrial usage of herbal plants has gone up, but techniques for verifying their botanical identity is still questionable. In the herbal industry, bulk consignments are received in powdered form as it is cumbersome to transport drugs in whole form. To ensure that the final product is safe and efficacious, the authenticity of the herbal plant should be established at the first stage. A proper methodology should be adopted in terms of computer technology to establish the correct botanical identity of the plant and to check the presence of substitutes and adulterants. Objective: To develop a software for identification of powdered samples of leaves and barks used in Ayurvedic Pharmacopoeia of India along with their substitutes and adulterants. Materials and Methods: Almost 100 plants have been selected from the Ayurvedic Pharmacopoeia of India comprising 54 barks and 46 leaves. Samples were self-collected and authenticated from the National Institute of Science Communication and Information Resources, Pusa, New Delhi. The selected crude herbal drugs were subjected to a detailed powdered microscopic identification and standard operating procedure for the preparation of slides was prepared. The features selected for identification of bark included14 specific characters - stone cells, calcium oxalate crystals, starch grains, medullary rays, fibers, sclereids, cork, isolated oil cells, tubular lactiferous canals, phloem parenchyma, masses, rhytidoma, parenchyma, and secretory canals. These characters are further differentiated into 75 features and 151 subfeatures, whereas for leaves, 13 specific characters were included, namely, epidermis, stomata, trichomes, calcium oxalate crystals, fibers, cell contents, cystoliths, lamina, starch grains, tracheids, lactiferous canals, and xylem vessels which are differentiated into 139 features. The details of all the features have been uploaded in the software under the name tool for identity of powdered herbals through analytical microscopy (www.tipham.com) with the database of 100 selected drugs. Results: A computer-based approach is developed which contains standard requirements for powdered plant parts, thus enabling identification of a bark or leaf powder in short time with minimum expertise. Conclusion: Computer-based technology would be a landmark in the field of pharmacognosy as proper identification of plant is the key to develop quality herbal products ensuring their safety and efficacy. Abbreviations used: μm: Micrometer; AHP: American Herbal Pharmacopoeia; DNA: Deoxyribonucleic acid; GMP: Good Manufacturing Practices; ICMR: Indian Council of Medical Research; Id: Identity Document; IT: Information Technology; MICROAID: Microaided Identification; MP: Megapixel; NA: Not Applicable; NISCAIR: National Institute of Science Communication and Information Resources; TIPHAM: Tool for Identity of Powdered Herbals through Analytical Microscopy; TLC: Thin-Layer Chromatography; UK: United Kingdom; WHO: World Health Organization.
  5,571 238 -
Carica Papaya leaf extract as a neuroprotective agent against behavioral and neurotransmitter changes in brain of the rat treated with sodium fluoride in Pre- and Post-Natal periods
Rajkiran Reddy Banala, Kiran Kumar Nagapuri, Khalid Pasha Mohd, M Manjula Reddy, Pratap Reddy Karnati
April-June 2018, 14(55):123-131
DOI:10.4103/pm.pm_378_17  
Background: Fluoride is an excitotoxin challenging the excitatory receptors and activate them continuously and also in proliferation of reactive oxygen and nitrogen species leading to neurodegeneration and its associated dysfunction. Chronic exposure to 20ppm sodium fluoride to pregnant rats is proven to be neurotoxic for the developing pups. Carica papaya is one of the medicinally important plants which exhibit anti-oxidative and anti-inflammatory responses. Objective: The present study aimed to evaluate the neuroprotective properties of Carica papaya leaf extract (CPLE) against fluoride induced behavioural change and neurotransmitter loss. Materials and methods: Timed pregnant Wistar rats (n=24) were chosen for the experiment and were divided into 4 groups (Group I: Control, Group II: 20 ppm Fluoride treated, Group III: 20 ppm Fluoride-50 mg CPLE, Group IV: 20 ppm Fluoride-100mg CPLE). The fluoride exposed groups received 20 ppm sodium fluoride (NaF) and two different concentrations of extract (50 and 100 mg/mL) through water for 52 days. The postnatal rat brains were collected after sacrificing them at regular time intervals (day 1, 7, 14, 21 and 30) and used to analyze the neurotransmitters alterations in cerebral cortex and hippocampus region of the brains. Results: Our results suggest that chronic exposure of NaF to pregnant rats and pups during embryonic and postnatal stages had detrimental effects on the brain. The concentration of fluoride increased in brain tissue and serum with respect to treatment duration in fluoride groups (groups 2, 3 and 4). The levels of glutamate and aspartate increased whereas the levels of acetylcholine, epinephrine, nor-epinephrine, dopamine and serotonin diminished on NaF exposure. The other neurotoxic effects of fluoride are behavioural changes including learning, motor control and other behavioural deficits in postnatal rats. Conclusion: The fluoride induced neurotransmitters and behavioural changes were ameliorated with CPLE administration, that were dose dependent i.e. 100 mg of CPLE showed better reversal efficiency than 50 mg extract.The present study reports the neuroprotective properties of Carica papaya leaf extract (CPLE) against fluoride-induced behavioral and neurotransmitter loss in developing rats. Day 1 pregnant Wistar rats (n = 24) were chosen as a model of study and were divided into four groups (Group I: control, Group II: 20 ppm fluoride treated, Group III: 20 ppm fluoride-50 mg CPLE, and Group IV: 20 ppm fluoride-100 mg CPLE). The fluoride-exposed groups received 20 ppm sodium fluoride (NaF) along with CPLE 50 or 100 mg/mL orally for 52 days. The brains of postnatal rats were collected at regular time intervals (day 1, 7, 14, 21, and 30) and used to analyze the neurotransmitters alterations in cerebral cortex and hippocampus region of the brain. The chronic exposure of NaF during embryonic and postnatal stages had detrimental effects on the brain. The levels of glutamate, aspartate, and epinephrine increased, whereas the levels of acetylcholine (Ach), norepinephrine, dopamine, and serotonin diminished on NaF exposure. Chronic fluoride exposure induced behavioral deficits (learning and motor control) in postnatal rats. The administration of CPLE extract has successfully ameliorated the fluoride-induced alterations in a dose-dependent manner, i.e. 100 mg of leaf extract showed better reversal efficiency than 50 mg extract. Abbreviations used: CPLE: Carica papaya leaf extract; NaF: Sodium fluoride; Ach: Acetylcholine; BSA: Bovine serum albumin; DA: Dopamine NE: Norepinephrine; EPN: Epinephrine; 5HT: Serotonin.
  4,073 160 -
A rapid high-performance liquid chromatography method for the simultaneous estimation of water-soluble vitamin in ten wild edible plants consumed by the tribal people of North-eastern Region in India
Tapan Seal, Kausik Chaudhuri, Basundhara Pillai
April-June 2018, 14(55):72-77
DOI:10.4103/pm.pm_481_17  
Background: Bauhinia purpurea, Clerodendrum colebrookianum, Dillenia pentagyna, Diplazium esculentum, Houttuynia cordata, Oenanthe linearis, Potentilla lineata, Perilla ocimoides, Sonchus arvensis, and Zanthoxylum acanthopodium are potent wild edible plants and consumed by the tribal people of North-eastern region in India. Objective: A reversed-phase high-performance liquid chromatographic method using photodiode array detector with gradient elution has been developed and validated for the simultaneous quantitation of several water-soluble vitamins such as ascorbic acid (Vitamin C), thiamine (Vitamin B1), riboflavin (Vitamin B2), niacin (Vitamin B3), pantothenic acid (Vitamin B5), pyridoxine (Vitamin B6), and folic acid (Vitamin B9) in these ten wild edible plants. Materials and Methods: The chromatographic separation of vitamins was carried out on Acclaim C 18 column (5 μm particle size, 250 mm × 4.6 mm), Dionex Ultimate 3000 liquid chromatograph and detection was carried out at three different wave lengths (210, 245, and 254 nm) using a mobile phase of acetonitrile and aqueous trifluoroacetic acid (0.01% v/v) solution with gradient elution. Conclusion: The results of investigation showed that these plants are rich sources of vitamins, especially the B group of vitamins that can contribute immensely to nutrition, food security, and health and therapeutic benefits. The high percentage of recovery (98%–99%), low coefficient of variation (R2 > 0.99), low limit of detection, and limit of quantitation confirm the suitability of the method for simultaneous quantification of vitamins in these ten plants under investigation. Abbreviations used: RSD: Relative standard deviation; LOD: Limit of detection; LOQ: limit of quantification; SEM: Standard error of mean; ND: Not detected; DPM: Dried plant material; TFA: Trifluoroacetic acid; v/v: Volume by volume; HPLC: High-performance liquid chromatography; nm: Nanometer; mg: Milligram; μg: Microgram; R2: Regression coefficient.
  3,811 242 -
Insulinotropic and cytoprotective effect of L-theanine: An in vitro dose dependent study
Arpita Saha, Sirshendu Chatterjee, Ananya Chatterjee, Surmi Roy, Nirmalya Roy, Satinath Mukhopadhyay, Sandip K Bandyopadhyay
April-June 2018, 14(55):36-39
DOI:10.4103/pm.pm_595_17  
Background: L-theanine is a natural bioactive amino acid present in tea (Camellia sinensis [L.] O. Kuntze) and consumed worldwide through tea decoction. Objective: The present work aims at evaluation of the insulin secretion and cytoprotective efficacy of L-theanine, a bioactive amino acid present in tea, on pancreatic β-cell line, RIN-m5F. Methods: RIN m5F cell were treated with L-theanine alone or prior to hydrogen peroxide (200 um) treatment and subsequently cell viability, cellular morphology, insulin secretion and insulin gene expression were analyzed. Results: Studies have shown that L-theanine dose dependently (0–300 μm) increases the β-cell mass as well as increases insulin production by RIN-m5F cell. It was also observed that pretreatment of the cell with L-theanine partially protected the oxidative stress of β-cells that were evident from cell viability, cellular morphology, and restoration of insulin-secreting ability. Conclusion: Results suggest that L theanine is an insulinotropic agent as well as effective in giving partial protection to pancreatic β-cells in oxidatively stressed condition. L-theanine can be used in the prevention or treatment of diabetes. Abbreviations used: EGCG: Epigallocatechin gallate; FBS: Fetal Bovine Serum; DMEM Dulbecco's Modified Eagle Medium
  3,663 154 -
Anticoagulant, antiplatelet and fibrin clot hydrolyzing activities of flax seed buffer extract
Sharath Kumar M. Nandish, Jayanna Kengaiah, Chethana Ramachandraiah, Ashwini Shivaiah, Chandramma , Kesturu S Girish, Kempaiah Kemparaju, Devaraja Sannaningaiah
April-June 2018, 14(55):175-183
DOI:10.4103/pm.pm_320_17  
Background: Flax seeds possess long array of medicinal qualities as it found to show beneficial effects on cardiovascular, hypertension, diabetes, cancer, inflammatory and autoimmune disorders. While, the therapeutic role of flax seed proteins on thrombotic disorder is least explored. Objective: The current study aims to identify the fibrin clot-dissolving, anticoagulant and antiplatelet properties of flaxseed buffer extract (FSBE) using Platelet Rich Plasma (PRP) and Platelet Poor Plasma (PPP). Materials and Methods: Flax Seed Buffer Extract (FSBE) proteins were characterized upon subjected to SDS-PAGE. The presence of cysteine protease in the extract was identified using colorimeter and zymography experiments. Anticoagulant activity was established using invitro recalcification time. While, the fibrinogenolytic, fibrinolytic and human plasma proteins degradation activities were proved using SDS-PAGE. Furthermore, antiplatelet activity was confirmed using Chronology dual channel whole blood/optical lumi aggregation system (Model-700). Toxicity studies were assayed using RBC cells and albino mice. Results: FSBE showed divergent protein banding pattern from the molecular mass ranging 200 kDa-14 kDa under both reduced and non-reduced conditions. The FSBE showed proteolytic activity as it hydrolyzed casein and gelatin with the specific activity 0.180 and 0.201 units/mg/min respectively. The proteolytic activity of FSBE was completely abolished only by IAA but PMSF, 1, 10 Phenanthroline and EDTA did not, revealing the presence of cysteine protease in the extract. FSBE showed anticoagulant activity as it enhanced the clotting time from control 220s to 320s. Furthermore, FSBE showed antiplatelet activity by inhibiting ADP and Epinephrine induced platelet aggregation and the observed aggregation inhibition was found to be 63.6% and 16.3% respectively. In addition, it hydrolyzed specifically Aα and Bβ chains of human fibrinogen and all the chains of human fibrin clot. The fibrinogenolytic activity was inhibited by IAA but PMSF, EDTA, 1, 10 Phenanthroline did not, suggesting the role of cysteine proteases. Interestingly, FSBE did not cause edema and hemorrhage in the experimental mice and did not hydrolyze RBC cells suggesting its nontoxic properties. Abbreviations used: FSBE: Flaxseed buffer extract; PMSF: Phenylmethylsulfonyl fluoride; IAA: Iodoacetic acid; EDTA: Ethylenediaminetetraacetic acid; PRP: Platelet-rich plasma; PPP: Platelet-poor plasma; INR: International normalized ratio; MED: Minimum edema dose; MHD: Minimum hemorrhagic dose; APTT: Activated partial thromboplastin time; PT: Prothrombin time; HMWK: High-molecular-weight kininogen.
  3,523 191 -
Lipid-lowering effect of hydroalcoholic extracts of Gynura procumbens in Chemical- and High-fat diet-induced hyperlipidemic rats
Vikneswaran Murugaiyah, Mohammed Ali Ahmed Saeed, Yow-Meng Kuong, Kisantini Murugesu, Subramani Parasuraman, Mohd. Zaini Asmawi, Amirin Sadikun
April-June 2018, 14(55):184-191
DOI:10.4103/pm.pm_451_17  
Objective: The present study was carried out to investigate lipid-lowering effect of hydroalcoholic extracts of Gynura procumbens in chemical- and high-fat diet (HFD)-induced hyperlipidemic rats. Materials and Methods: The lipid-lowering effect of hydroalcoholic extracts was investigated in poloxamer-407 (P-407)-induced acute hyperlipidemic rat model. The most active extract was subjected to phytochemical analysis by high-performance liquid chromatography-ultraviolet (HPLC-UV) using phenolic acids as marker compounds and evaluated in HFD-induced chronic hyperlipidemic rats. Results: The evaluation on hydroalcoholic extracts of G. procumbens showed that 95% ethanolic extract (95EEGP) was the most potent extract that significantly reduced serum total cholesterol (TC, P < 0.05) and triglycerides (TG, P < 0.001) levels. The 95EEGP contained 7.18, 3.20, 28.31, and 9.72 mg/g dried extract of chlorogenic acid (CA), 3,4-dicaffeoylquinic acid (3,4DC), 3,5DC, and 4,5DC, respectively. The extract at doses of 200 and 500 mg/kg significantly reduced serum TC, TG, low-density lipoprotein-cholesterol (LDL-C), and atherogenic index (A. I.) levels of the P-407-induced hyperlipidemic rats, in a dose-dependent manner (P < 0.01 or better) but had no effect on high-density lipoprotein-cholesterol (HDL-C). On HFD-induced hyperlipidemic rats, the 95EEGP at doses of 250, 500, and 1000 mg/kg significantly reduced the serum TC, TG, LDL-C, and A. I. levels (P < 0.05 or better) while increased serum HDL-C (P < 0.001). The effect was dose-dependent showing comparable effect to that of atorvastatin at moderate and high doses. Conclusion: The present study demonstrated the lipid-lowering potential of the 95EEGP of G. procumbens in chemical- and HFD-induced hyperlipidemic rat models. Further investigations are warranted to elucidate its mechanism of its lipid-lowering action. Abbreviations used: HFD, high-fat diet; P-407, poloxamer-407; 95EEGP, 95% ethanolic extract; TC, total cholesterol; TG, triglycerides; 3,4DC, 3,4-dicaffeoylquinic acid; 3,5DC, 3,5-dicaffeoylquinic acid; 4,5DC, 4,5-dicaffeoylquinic acid; LDL-C, low-density lipoprotein-cholesterol; A. I., atherogenic index; HDL-C, high-density lipoprotein-cholesterol; CVD, cardiovascular disease; HMG CoA, 3-hydroxy-3-methylglutaryl coenzyme A; G. procumbens, Gynura procumbens; HPLC-UV, high-performance liquid chromatography-ultraviolet; CA, chlorogenic acid; 75EEGP, 75% ethanolic extract; 50EEGP, 50% ethanolic extract; 25EEGP, 25% ethanolic extract; AEGP, water extract; MeOH, methanol; SEM, standard error mean; VLDL-C: Very low-density lipoprotein-cholesterol.
  3,039 176 -
Formulation design and evaluation of an emulgel containing Terminalia arjuna bark extract for transdermal delivery
Dinanath Tukaram Gaikwad, Namdeo Ramhari Jadhav
April-June 2018, 14(55):249-255
DOI:10.4103/pm.pm_578_17  
Background: Terminalia arjuna is used in various indigenous system of medicine such as Ayurveda, Siddha, and Unani. The traditional medical forms provide drug delivery with peaks often above the required dose and face problems such as first-pass metabolism, stability, and low therapeutic efficacy. To enhance topical delivery and to allow for controlled release, the use of T. arjuna bark extract emulgel was explored. Objective: The objective of this study is to design and develop novel emulgel formulation of T. arjuna bark extract for transdermal delivery. Materials and Methods: Pseudoternary phase diagram was developed using aqueous titration method. Appropriate amount of oil, surfactant, and cosurfactant were mixed together in accordance with obtained emulsion region in the phase diagram. The emulsion was prepared and incorporated in gel base. The four different formulations were evaluated for physical examinations, rheological studies, skin irritation studies, in vitro release, and ex vivo release studies. Results: The phase diagram at Km value 3 showed better emulsion existence regions. Formulation F3 showed maximum release, wherein the amount of the extract released after 720 min was 91.43%. Skin irritation test on albino rats resulted no allergic symptoms such as inflammation, redness, and irritation up to 72 h. All the prepared emulgel formulations were found to be stable on storage. Conclusion: It can be concluded that T. arjuna bark extract emulgel can be used for transdermal delivery for the treatment of chronic ailments such as pulmonary hypertension.
  3,039 174 -
Alliin the precursor of allicin in garlic extract mitigates proliferation of gastric adenocarcinoma cells by modulating apoptosis
Debjani P Mansingh, Nibedita Dalpati, Veeresh Kumar Sali, A Hannah Rachel Vasanthi
April-June 2018, 14(55):84-91
DOI:10.4103/pm.pm_342_17  
Background: Garlic, a common spice used since time immemorial for various purposes, is considered a potential functional food as it exhibits cardioprotection to chemoprevention properties due to the presence of organosulfur constituents in each garlic pod. Alliin is not widely studied for its bioactivity as it is an unstable compound which is converted to allicin on mechanical and chemical degradation. Objective: Hence, in the present study, the influence of alliin on gastric adenocarcinoma (AGS) cells and normal intestinal cells (INT-407) would be tested for its potent antiproliferative effect and mechanism of action. Materials and Methods: The quantity of alliin in fresh and dried garlic extract was measured by high-performance liquid chromatography to corroborate the cytotoxicity activity identified by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay and acridine orange/ethidium bromide staining. Further, to identify the mechanism of action, DNA fragmentation assay, annexin V Assay, and flow cytometry analysis of apoptosis followed by expression of apoptotic proteins such as Bax, Bcl-2, and cytochrome-C by Western blot was done. Results: It was identified that alliin inhibited proliferation of gastric carcinoma cells by decreasing the cell viability but not in the normal intestinal cells. The level of apoptosis was modulated by reactive oxygen species generation and decrease in mitochondrial membrane potential mediated by deregulation of Bax/Bcl-2 level at protein level leading to upregulation of Cytochrome C. Conclusion: It is impressive to note that alliin content was high in fresh aqueous extract compared to that of dried garlic extract which concludes that the use of garlic from time immemorial is a worthy functional food in its fresh form to combat cancer cells. However, in-vivo studies are warranted. Abbreviations used: AGS-Gastric adenocarcinoma; ROS-Reactive oxygen species; MMP-Mitochondrial membrane potential; TBS- Tris buffered saline.
  2,869 201 -
Antioxidant, anti-inflammatory, and antiproliferative activity of extracts obtained from Tabebuia Rosea (Bertol.) DC
Francisco Javier Jimenez-Gonzalez, Jenny Marcela Vélez-Gómez, Jhon Jairo Melchor-Moncada, Luz Angela Veloza, Juan Carlos Sepúlveda-Arias
April-June 2018, 14(55):25-31
DOI:10.4103/pm.pm_624_17  
Background: Tabebuia rosea (Bertol.) DC. is a neotropical tree used in traditional medicine in the Northern coast of Colombia as well as Latin America for infectious diseases treatment. Few studies have evaluated the biological activity of this species. Objective: The objective of this study is to determine the antioxidant, anti-inflammatory, and antiproliferative potential of leaf and inner bark extracts from T. rosea. Materials and Methods: The antioxidant activity was determined using the 2,2-diphenyl-1-picrylhydrazyl (DPPH) and oxygen radical absorbance capacity (ORAC) methods. The anti-inflammatory activity was evaluated in lipopolysaccharide-stimulated murine macrophages. In vitro antiproliferative effect was determined in HepG2, HeLa, MCF-7, and B16F10 cell lines. Results: The highest DPPH radical scavenging activity was observed for T. rosea ethyl acetate leaf extract (IC50of 157.5 ± 2.4 μg/mL). This extract also induced the best antioxidant activity as determined by ORAC (11,112.2 ± 1,255.3 μmol TE/g of extract). Moreover, T. rosea leaf n-hexane, chloroform, and aqueous extracts, in addition to inner bark aqueous extract did inhibit nitric oxide production by over 90%. In addition, inner bark extracts markedly inhibited prostaglandins E2 and tumor necrosis factor alpha (>90%). The best antiproliferative activity was displayed by the inner bark chloroform extract against HepG2 (selectivity index [SI] = 5.50) and B16F10 (SI = 3.18) cell lines. Conclusion: These results demonstrate the potential biological activity of T. rosea extracts. Abbreviations used: DPPH: 2,2-diphenyl-1-picrylhydrazyl; ORAC: Oxygen radical absorbance capacity; LPS: Lipopolysaccharide; NO: Nitric oxide; TNF-a: Tumor Necrosis Factor Alpha; PGE2: Prostaglandin E2; TAC: Total antioxidant content; MTT: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide.
  2,861 159 -
Ameliorative effect of quercetin and rutin via modulation of hypothalamic–Pituitary–Adrenal axis and regulation of fasting glucose in chronic stress-induced prediabetes
Mustajab Quraishi, Santosh N Mokale, Nikhil S Sakle
April-June 2018, 14(55):65-71
DOI:10.4103/pm.pm_323_17  
Background: Quercetin (QUE) and Rutin (RUT) have nutritive and medicinal values. On the other hand, there are no reports of scientific assessment of its hypothalamo–pituitary–adrenal (HPA) axis modulation in the treatment of prediabetes (DM). Aim: The current study was designed to investigate the modulatory effects of QUE, RUT, and escitalopram (ESC) as antidepressants on HPA axis in chronic stress-induced pre-DM in rats. Materials and Methods: The experimental protocol was of 5 weeks. Chronic unpredictable mild stress (CUMS) was used as a model of depression to induce pre-DM in rats. The treatment was started at the end of 4th week. After 5th week, the plasma adrenocorticotropic hormone (ACTH), serum corticosterone (CORT), fasting blood glucose (FBG), and behavioral parameters were evaluated. Results: Oral administration of QUE (50 mg/kg), RUT (50 mg/kg), and ESC (2.5 mg/kg) to stressed control alleviated HPA axis-associated parameters (ACTH and CORT) and significantly decreased the FBG. Besides this, the depressive effects induced by CUMS were significantly improved as evident from results indicating a promising antidepressant activity. Moreover, submaximal dose of QUE (25 mg/kg) and RUT (25 mg/kg) enhanced the antidepressant activity of ESC (1 mg/kg, p.o.), which suggests that they may act through the HPA axis. Conclusion: Current results suggest that chronic stress in rats causes dysregulation of the HPA axis which leads to diabetic-like condition, i.e., “pre-DM.” It is possible that QUE, RUT, and ESC may be able to suppress the HPA axis response which could be beneficial for the treatment of stressed diabetic patients. Abbreviations used: QUE: Quercetin; RUT: Rutin; HPA: Hypothalamic–Pituitary–Adrenal Axis; CUMS: Chronic unpredictable mild stress; ACTH: Adrenocorticotropic hormone; CORT: Corticosterone; FBG: Fasting Blood Glucose; ESC: Escitalopram; FST: Fasting Blood Glucose.
  2,714 277 -
Molecular interaction of naringin and its metabolite naringenin to human liver fibrosis proteins: An In Silico approach
VJ Shine, GI Anuja, S Pradeep, SR Suja
April-June 2018, 14(55):102-109
DOI:10.4103/pm.pm_453_17  
Background: Naringin, pharmaceutically active flavonoid, rapidly metabolizes in liver into naringenin. Both naringin and naringenin have significant biological activity and less toxicity. Objective: In the present study, in silico molecular interactions of naringin and its metabolite naringenin have been evaluated against different human liver fibrosis proteins. Materials and Methods: The major human therapeutic protein targets such as epidermal growth factor receptor (EGFR), vascular endothelial growth factor receptor-2 (VEGFR-2), fibroblast growth factor receptor-1 (FGFR1), Kelch-like ECH-associated protein-1 (Kaep1), transforming growth factor beta receptor I (TGFBR-1), angiotensin II receptor type-1 (Angio-II-Type-1), Janus kinase-2 (JAK-2), Zeta-chain-associated protein kinase-70 (ZAP-70) have been selected for the docking studies. This computational study was performed using Schrödinger Suite Maestro 10.3 Glide software 2015. Results: The studies demonstrated comparable binding affinities of naringin and naringenin with human therapeutic protein targets such as JAK-2, ZAP-70 Kinase, Angio-II-Type 1, TGFBR1, Kaep1, EGFR, VEGFR-2, and FGFR1 when compared to their respective standard drugs such as gefitinib, regorafenib, dovitinib, bardoxolone methyl, SB-431542, olmesartan, and ruxolitinib. Naringin showed better glide score ranging from −8.5 to −13.3 kcal/mol whereas its metabolite Naringenin also showed comparable glide score ranging from −5.4 to −9.3 kcal/mol. The binding of target proteins with respective standard drugs showed −2.2 to −10.12 kcal/mol. Conclusion: The observed in silico human protein interactions of naringin and its metabolite naringenin could be exploited for the anti-liver fibrosis therapy. The results derived from this pioneering virtual study may advance further mechanistic in vitro and preclinical in vivo studies. Abbreviations used: Jak-2: Janus kinase-2, ZAP-70: Zeta-chain-associated protein kinase-70, Angio-II-type-1: Angiotensin II receptor type-1, TGFBR1: Transforming growth factor beta receptor I, Kaep1: Kelch-like ECH-associated protein-1, EGFR: Epidermal growth factor receptor, VEGFR-2: Vascular endothelial growth factor receptor-2, FGFR1 kinase: Fibroblast growth factor receptor-1.
  2,491 155 -
Hydroalcoholic extract of Argyreia speciosa roots ameliorates HCl-mediated acute lung injury in mice
Gurparteek Kaur, Priya Jaswal, Reenu Banga, Vivek Dharwal, Ashwani Kumar, Amarjit Singh Naura
April-June 2018, 14(55):8-13
DOI:10.4103/pm.pm_324_17  
Objective: Argyreia speciosa is a popular Indian medicinal plant. It has long been used in the traditional Ayurvedic Indian medicine for various diseases. The present work was designed to evaluate anti-inflammatory potential of hydroalcoholic extract of A. speciosa roots using mouse model of HCl-induced acute lung injury (ALI) by conducting a dose–response studies. Materials and Methods: The extract was given once orally at a dose of 50, 100, or 200 mg/kg b.wt. 90 min before HCl administration. Lung tissue of different group of mice was analyzed for the recruitment of inflammatory cells, redox status, and mRNA expression of pro-inflammatory genes to evaluate the anti-inflammatory effects of the extract. Results: Our results showed that the extract reduced HCl-induced lung inflammation in a dose-dependent manner. The neutrophil numbers in bronchoalveolar lavage fluid (BALF) were almost completely abrogated at a dose of 100 mg/kg b.wt. The severe reduction in neutrophils was accompanied with reduced pulmonary edema as the total protein content in BALF was found to be decreased substantially. In addition, the extract prevented the HCl-mediated oxidative stress in lungs as reflected by the normalization of levels of reactive oxygen species, malondialdehyde, redox status, and catalase activity. Extract seems to blunt the activity of redox-sensitive transcription factor nuclear factor-kappa B (NF-κB) as it suppressed the gene expression of ALI-linked pro-inflammatory cytokines, namely, interleukin-1 β and tumor necrosis factor-alpha. Conclusion: Overall, our data provide evidence that cocktail of natural phytochemicals present in hydroalcoholic extract of A. speciosa protects against HCl-induced ALI in mice potentially by curbing oxidative stress and consequent activation of NF-κB in the tissue. Abbreviations used: ALI: Acute lung injury, BALF: Bronchoalveolar lavage fluid, IL-1: Interleukin-I, TNF-a: Tumor necrosis factor-alfa, ROS: Reactive oxygen species, MDA: Malondialdehyde
  2,487 123 -
Evaluation and understanding the molecular basis of the antimethicillin-resistant Staphylococcus aureus activity of secondary metabolites isolated from Lamium amplexicaule
Mohammed M Ghoneim, Arafa Musa, Atef A El-Hela, Khaled M Elokely
April-June 2018, 14(55):3-7
DOI:10.4103/pm.pm_541_17  
Background: The genus Lamium includes about forty annual or perennial plants distributed everywhere, it has significant biological activities including antimicrobial, antioxidant and antischistosomal effects. However, no detailed reports about the antimicrobial (Anti-MRSA) effect of the isolated metabolites. Objective: Studying the mechanism of action of the antimicrobial (Anti-MRSA) activity of the isolated metabolites. Materials and Methods: The EtOAc extract of L. amplexicaule was subjected to different chromatographic methods to isolate the secondary metabolites, and the isolated compounds were elucidated by spectroscopic techniques. The antimicrobial activity against strains of microorganisms was performed according to Minimum Inhibitory Concentration, the study of Anti-MRSA activity was explained by molecular docking against CrtM enzyme. Results: Phytochemical study of the aerial parts of L. amplexicaule resulted in the isolation of 5 known compounds; phytol (1), b-sitosterol (2), isorhamnetin (3), 3,4-dihydroxy-methyl benzoate (4), and hydroxynervonic acid (5). The antimicrobial activity of the isolated metabolites revealed that compounds 1, 3, and 4 have pronounced antimethicillin-resistant Staphylococcus aureus (MRSA) effect. Conclusion: These all known compounds were firstly isolated from L. amplexicaule. Three of them showed pronounced anti- MRSA effect, The mechanism of action against dehydrosqualene synthase enzyme was established. In addition, the study of molecular determinates of activity of these new scaffolds as anti-MRSA has a great importance for the development of new anti-MRSA candidates. Abbreviations used: L. amplexicaule: Lamium amplexicaule; CrtM: Dehydrosqualene synthase; MRSA: Methicillin-resistant strains of S. aureus; STX: Staphyloxanthin; UV: Ultraviolet-visible; TLC: Thin-layer chromatography; ESI/MS: Electrospray mass spectrometry; VLC: Vacuum liquid chromatography; L: Liter
  2,439 145 -
Prophylactic treatment with icariin prevents isoproterenol-induced myocardial oxidative stress via nuclear Factor-Like 2 activation
Sumit Sharma, Vasim Khan, Abul Kalam Najmi, Ozair Alam, Syed Ehtaishamul Haque
April-June 2018, 14(55):227-236
DOI:10.4103/pm.pm_469_17  
Introduction: Icariin, a major component of Epimedium species and a mild phosphodiesterase 5 (PDE-5) inhibitor, was evaluated for the prevention of myocardial oxidative stress in isoproterenol (ISO)-challenged Wistar rats. Objective: This study aimed to evaluate the cardioprotective action of icariin in ISO-intoxicated rats. Materials and Methods: Rats were daily treated with icariin (1, 5, and 10 mg/kg, p.o.) and sildenafil (0.7 mg/kg, i.p.) for 15 days. Oxidative stress was induced by subcutaneous administration of ISO (85 mg/kg s.c) in two consecutive doses at an interval of 24 h on 14th and 15th day of the study. After induction, rats were anesthetized for recording the electrocardiogram (ECG) and then sacrificed to perform immunohistochemistry and biochemical assays of heart tissue. Results: ISO treatment resulted in a marked increase in lipid peroxidation, serum markers (lactate dehydrogenase [LDH], creatine kinase-MB [CK-MB], and C-reactive protein [CRP]), and infarct size and a significant decrease in the level of reduced glutathione (GSH) and endogenous antioxidant enzymes in the myocardium. Lowering of arterial blood pressure and alteration in ECG showed significant alteration in cardiac hemodynamics. Hematoxylin and eosin staining of the cardiac tissue showed considerable myocardial damage. Pretreatment with icariin (5 and 10 mg/kg, p.o.) and sildenafil (0.7 mg/kg, i.p) significantly decreased the elevated lipid peroxidation, LDH, CK-MB, and CRP. Moreover, the results also showed an increase in endogenous antioxidants and protein expression of nuclear factor-like 2 (Nrf-2) when compared to the ISO-treated group. Conclusion: The results indicated that icariin significantly ameliorates the ISO-induced oxidative stress and restores membrane integrity and cellular damage. Thus, we can conclude that the activation of Nrf-2 signaling and PDE-5 inhibition by icariin is possibly responsible for the cardioprotection. Abbreviations used: Nrf-2: Nuclear factor-like 2, PDE-5: Phosphodiesterase-5, ISO: Isoproterenol, TCM: Traditional Chinese Medicine, ECG: Electrocardiogram, LDH: Lactate dehydrogenase, CK-MB: Creatine kinase-MB, CRP: C-reactive protein, GSH: Endogenous glutathione, TBARS: Thiobarbituric acid reactive substances, GPx: Glutathione peroxidase, GR: Glutathione reductase, GST: Glutathione-S-transferase, SOD: Superoxide dismutase, CAT: Catalase, H and E: Hemotoxylin and eosin, MAPK: Mitogen-activated protein kinase, CPCSEA: Committee for the Purpose of Control and Supervision of Experiments on Animals guidelines, DMSO: Dimethyl sulfoxide, MM-GBSA: Molecular mechanics-generalized born surface area, BPM: Beats per minute.
  2,410 149 -
Effects of beta-sitosterol on isolated human non-pregnant uterus in comparison to prostaglandin E2
Cristina Occhiuto, Domenico Trombetta, Antonella Smeriglio, Emanuele Sturlese, Francesco Occhiuto
April-June 2018, 14(55):118-122
DOI:10.4103/pm.pm_163_17  
Background: Beta-sitosterol (β-sitosterol) is one of the several phytosterols widely studied for its potential to reduce benign prostatic hyperplasia and blood cholesterol levels. Objective: In the present study, the effects of β-sitosterol on spontaneous and agonist-induced contractions in in vitro nonpregnant human uterus with respect to prostaglandin E2(PGE2) were investigated. Materials and Methods: Myometrial strips, measuring approximately 15 mm × 4 mm × 2 mm, were attained from hysterectomy samples of premenopausal women. Longitudinal muscle strips were mounted on tissue baths, under physiological conditions, to measure their isometric contraction. The effects of cumulative amounts of β-sitosterol on spontaneous motility in the absence and presence of prazosin, atropine, fulvestran, indomethacin, or ethylenediaminetetraacetic acid (EDTA), and on agonist-induced motor activity, were examined. Results: On strips in the follicular phase, both β-sitosterol (1–100 μg/ml) and PGE2(0.1–10 μg/ml) increase, in a concentration-dependent manner, muscular basic tonus and amplitude and frequency of spontaneous uterine contractions; whereas on strips obtained during periovulatory phase, β-sitosterol and PGE2cause inhibition of uterine motility. For contractile response, the effective concentrations (EC50) were 47.8 μg/ml and 5.19 μg/ml, respectively. Unlike indomethacin, the tissue pretreatment with prazosin, fulvestran, atropine, or ethylenediaminetetraacetic acid did not affect the contractile uterine responses to β-sitosterol. Furthermore, the β-sitosterol was able to potentiate the contractile response induced by acetylcholine and vasopressin. Conclusions: These observations suggest that β-sitosterol may be a useful modulator of the uterine motility during menstrual cycle, facilitating female fertility.
  2,385 89 -
Naringin ameliorates doxorubicin-induced neurotoxicity In vitro and cognitive dysfunction In vivo
Grandhi Venkata Ramalingayya, Pawan G Nayak, Rekha R Shenoy, Sanchari Basu Mallik, Karthik Gourishetti, Shalam M Hussain, Chamallamudi Mallikarjuna Rao, Krishnadas Nandakumar
April-June 2018, 14(55):197-207
DOI:10.4103/pm.pm_364_17  
Objectives: The primary objective of the study was to study the neuroprotective potential of naringin (NAR) against doxorubicin (DOX)-induced neurotoxicity in vitro and DOX-induced cognitive deficits (chemobrain) in vivo. Materials and Methods: In vitro methods, viz., 3-[4,5dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide assay, flow cytometry, acridine orange/ethidium bromide staining, and neuritogenic and reactive oxygen species (ROS) assays, assessed neuroprotective potential of NAR and its aglycone naringenin (NGN) in IMR-32 cells. Chemobrain was developed in Wistar rats on chronic administration of ten cycles of DOX, and episodic memory was assessed using novel object recognition task. Serum cortisol, locomotor activity, and hematological biochemical and histological analysis were carried out. Results: A protective effect of NAR or NGN was observed upon pretreatment with the respective compounds in IMR-32 cells challenged with DOX. Flow cytometry revealed that flavonoids reduced cell cycle changes produced by DOX. In addition, an increase in apoptosis, intracellular ROS generation, and inhibition of neurite growth was noticed in IMR-32 cells with DOX treatment, which was significantly prevented by NAR or NGN pretreatment. Interestingly, NAR (50 mg/kg, p.o.) significantly ameliorated episodic memory deficit associated with DOX without influencing locomotion, upon chronic treatment. NAR also prevented histological changes to major organs observed with DOX. Conclusion: NAR showed neuroprotective potential and may be used as an adjuvant therapy for amelioration of neurocognitive complications associated with chemotherapy in cancer survivors. Abbreviations used: CKL: Creatine kinase level; COX: Cyclooxygenase; DMEM: Dulbecco's modified eagle media; DOX: Doxorubicin; FBS: Fetal bovine serum; MTT: 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide; NAR: Naringin; NGN: Naringenin; NORT: Novel object recognition task; NOS: Nitric oxide synthase; QOL: Quality of life; RA: retinoic acid.
  2,334 139 -
Molecular authentication of Trichosanthes species traded as “Patola:” An ayurvedic drug resource
Bhagyashri Kumbhalkar, Anuradha Upadhye, Shubhada Tamhankar
April-June 2018, 14(55):52-56
DOI:10.4103/pm.pm_242_17  
Background: Different interpretations of ayurvedic texts have recognized/recommended taxonomically distinct species/varieties of Trichosanthes as “Patola,” which is used extensively in the Indian traditional system of medicine for curing a variety of disorders. Sources of “Patola” also vary depending on region-specific availability and diverse traditions in different parts of India, which creates ambiguity in correct identity of “Patola.” Since “Patola” is one of the medicinal plant species traded in high volumes, its exact taxonomic identity and authentication is very essential. Objectives: Molecular profiling of different taxa recognized as “Patola” was carried out for correct identification and authentication of commercial crude drug samples. Materials and Methods: Molecular marker profiles of different taxa recognized as “Patola” were generated using inter-simple sequence repeat (ISSR) markers. Attempt was also made to authenticate commercial crude drug samples, using ISSR markers. Results: Selected 10 polymorphic ISSR primers generated distinct amplification profiles for the taxa traded as “Patola.” Moreover, two varieties of Trichosanthes cucumerina (var. cucumerina and var. anguina) could be clearly distinguished based on the specific bands produced by primers UBC 812, 822, 887, 888, and 889. Molecular profiling of market samples along with the genuine plant samples confirmed the trade of different Trichosanthes species as well as Momordica charantia under the name of “Patola.” Conclusion: To our knowledge, this is a first report on the application of ISSR markers for identification and authentication of “Patola.” Developed DNA profiles should be useful in investigations of adulteration in market samples and establishment of correct identity. Abbreviations used: AFLP: Amplified fragment length polymorphism; AHMA: Agharkar Herbarium at Maharashtra Association; DNA: Deoxyribonucleic acid; ISSR: Inter-simple sequence repeat; MC: Momordica charantia; PCR: Polymerase chain reaction; RAPD: Random amplified polymorphic DNA; TAE: Tris acetate-EDTA buffer; TC: Trichosanthes cucumerina; TCA: Trichosanthes cucumerina var. anguina; TCC: Trichosanthes cucumerina var. cucumerina; TD: Trichosanthes dioica; TT: Trichosanthes tricuspidata; TE: Tris-EDTA buffer; UBC: University of british Columbia, Vancouver, Canada.
  2,357 115 -
Comparative effects of metformin, Pleurotus ostreatus, Nigella Sativa, and Zingiber officinale on the streptozotocin-induced diabetes mellitus in rats
Sibghatullah Muhammad Ali Sangi, Abdulhakim Bawadekji, Mouhanad Al Ali
April-June 2018, 14(55):268-273
DOI:10.4103/pm.pm_108_18  
Background: Diabetes mellitus (DM) is a heterogeneous metabolic disorder occurring due to absolute or relative deficiency of insulin and causing chronic hyperglycemia that leads to crippling complications associated with morbidity and mortality. Objectives: The objective of the present work is to investigate the comparative antihyperglycemic effects of Pleurotus ostreatus, Nigella sativa, and Zingiber officinale with metformin. Materials and Methods: The study was conducted from June 2017 to February 2018. In this study, rats were divided into six groups according to the treatment regimen. Each group comprised five rats. Serum analysis of glucose was conducted during the treatment. After animals were sacrificed, histopathological examination of the pancreas was carried out. Results: Histopathological examination showed that streptozotocin caused damage to pancreatic beta-cells which secrete the insulin. Treatment with metformin, N. sativa, mushroom, and ginger produced a significant improvement in the function of these cells, regeneration of cells, and decrease in serum glucose levels. Conclusion: In two groups, N. sativa and ginger, regeneration of beta islets was observed, along with a significant reduction in the serum glucose levels, which was found in other groups as well.
  2,232 165 -
Nontargeted analysis and cancer cells cytotoxicity of Aegle marmelos Correa Ex Roxb.
Sachin Kumar, Ramesh Babu Bodla
April-June 2018, 14(55):40-44
DOI:10.4103/pm.pm_264_17  
Background: Aegle marmelos belongs to the family Rutaceae, a medium-sized perennial tree which grows in subtropical and tropical parts of India and South-East Asia. Studies have reported that leaf extracts of A. marmelos have anticancer, cardiotonic, antidyslipidemic, and hypoglycemic effects and were ethnologically used for the treatment of dropsy, opthalmitis, ulcers, beri beri, and cholera. Objective: In the present study, we have carried out nontargeted analysis of the leaf extract of A. marmelos Correa ex Roxb., by gas chromatography-mass spectrometry (GC/MS) and estimated the inhibitory concentration (IC50) in human cancer cell lines using MCF-7, H-460, and SF-268. Materials and Methods: Dried leaves of A. marmelos were extracted with 50% aqueous alcohol using Soxhlet apparatus. The dried extract was suspended in water and re-extracted with petroleum ether and the fraction was named as AMPEF (petroleum ether fraction of Aegle marmelos leaf extract); the dry yield of the AMPEF was found to be 7.56% (w/w). Nontargeted GC/MS analysis of AMPEF was performed using Shimadzu QP 2000 GC equipped with ULBON-HR-5 capillary column and mass spectrometer as detector. The cancer cell lines were obtained from the National Centre for Cell Science, Pune. In vitro cytotoxic activity of AMPEF on cancer cell lines was conducted using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium assay. Results: GC/MS analysis of the AMPEF enabled the identification of 26 compounds, and IC50of AMPEF in MCF-7, H-460, and SF-268 was found to be 0.53 ± 0.10, 0.65 ± 0.05, and 0.18 ± 0.01 μg/ml, respectively. Conclusion: The AMPEF was found to be a potential cytotoxic agent against the used cell lines. Abbreviations used: AMPEF: Petroleum ether fraction of leaf extract of Aegle marmelos; DMBA: 7,12-Dimethylbenz[a]anthracene; IC: Inhibitory concentration; MTT:3-[4,5-dimethylthiazole-2-yl]-2,5-diphenyltetrazolium bromide; NIST: National Institute of Standards and Technology; RT: Retention time; TIC: Total ion chromatogram.
  2,155 182 -
Evaluation of aphrodisiac activity of AHPL/AYCAP/0114 capsule in sexually sluggish male rats
Sanjay U Nipanikar, Dheeraj H Nagore, Sohan S Chitlange
April-June 2018, 14(55):264-267
DOI:10.4103/pm.pm_363_17  
Background: Male sexual dysfunction includes a wide range of problems such as erectile dysfunction and premature ejaculation, for which medical help is sought. The conventional use of various pharmacological and nonpharmacological measures are associated with multiple adverse events. Hence, a polyherbal formulation AHPL/AYCAP/0114 capsule was developed to provide safe and effective therapeutic option. Objectives: The aim of this study was to evaluate the aphrodisiac potential of AHPL/AYCAP/0114 capsule by assessing sexual behavior parameters such as mount frequency (MF), mount latency (ML), intromission frequency (IF), intromission latency (IL), ejaculation latency (EL), and ejaculation frequency (EF) in sexually sluggish male rats. Materials and Methods: Male Wistar rats were divided into six groups (n = 6) and treated with testosterone (30 mg/kg), sildenafil citrate (0.5 mg/kg), and AHPL/AYCAP/0114 capsule (110, 220, and 440 mg/kg) as Group II, Group III, and Group IV, V, and VI, respectively, for 28 days. Group I animals served as control. Then, they were introduced to sexually active female rats and their sexual behavior was recorded. Results: A significant increase in MF, IF, and EF was observed in all the AHPL/AYCAP/0114 capsule groups as compared to control group. A significant decrease in ML, IL, and EL was observed in all the AHPL/AYCAP/0114 capsule groups as compared to control group. AHPL/AYCAP/0114 capsule was well tolerated by rats. Discussion: Increase in MF, IF, and EF and reduction in ML, IL, and EL are indicative of increased sexual motivation and arousal and prolonged duration of coitus. This suggests enhanced sexual performance in sexually sluggish animals treated with AHPL/AYCAP/0114 capsule. Conclusion: It can be concluded that AHPL/AYCAP/0114 capsule possesses aphrodisiac activity and can be used in erectile dysfunction and loss of libido. Abbreviations used: (MF): Mount frequency; (ML): Mount latency; (IF): Intromission frequency; (IL): Intromission latency; (EL): Ejaculation latency; (EF): Ejaculation frequency; (ED): Erectile dysfunction; CPCSEA: Committee for the Purpose of Control And Supervision of Experiments on Animals.
  2,176 136 -
A therapeutic approach to target mitochondrial dysfunction using molecular docking studies: Screening of natural drugs for oral carcinoma
Manish Singh, Manish Kumar Tripathi, Alok Kumar Singh, Chandra Shekhar Azad, Indrajeet Singh Gambhir, Brijesh Kumar, Suresh Purohit
April-June 2018, 14(55):192-196
DOI:10.4103/pm.pm_471_17  
Background: Mitochondrial dysfunction is the major cause of various types of cancer, leading to death worldwide. The present study investigated the in silico binding potential of natural flavonoids and essential oils with human cyclophilin D (CyPD) protein. CyPD protein is a major molecular marker for apoptosis and has been reported to be elevated in oral carcinoma. Methods: PubChem database was used to check the efficacy of different active phytoconstituents (kaempferol, quercetin, eugenol, oxyresveratrol, tanshinone 2a, catechin, epicatechin, cinnamaldehyde, and emodin). These compounds were used as ligands to check their potential as anticancer agents against the inner mitochondrial membrane protein, CyPD. Docking studies were performed with the help of Discovery Studio 2.5 and Autodock. Emodin was used as a reference inhibitor to compare the results. Results: The binding energy (B.E.) of the reference inhibitor (known/established drug) emodin was observed −28.9 kcal/mol while novel inhibitors (catechin, cinnamaldehyde, epicatechin, eugenol, kaempferol, oxyresveratrol, quercetin, and tanshinone 2a) exhibited a range from −51.51 to −5.89 kcal/mol. Quercetin, kaempferol, and epicatechin (B.E.: −51.51, −34.79, and −30.62 kcal/mol, respectively) showed strong affinity as compared to reference inhibitor (B.E.: −28.9 kcal/mol). Conclusion: Quercetin, kaempferol, and epicatechin can be used as lead inhibitors against targeting CyPD. Abbreviations used: CyPD: Cyclophilin D, BE: Binding Energy, PTPC: Permeability transition pore complex, mPTP: Mitochondrial permeability transition pore.
  2,151 152 -
Therapeutic potential of Boerhavia diffusa L. against cyclosporine A-Induced mitochondrial dysfunction and apoptosis in madin–Darby canine kidney cells
MK Kalaivani, Sumathy Arockiasamy, Cordelia John, Hannah Rachel Vasanthi, P Soundararajan
April-June 2018, 14(55):132-140
DOI:10.4103/pm.pm_383_17  
Introduction: Cyclosporine A (CsA), a calcineurin inhibitor, causes chronic nephrotoxicity during organ transplantation. Objective: The aim is to investigate the effect of ethyl acetate fraction (EF) of Boerhavia diffusa on CsA-induced cell damage due to apoptosis and reactive oxygen species (ROS). Materials and Methods: Madin–Darby Canine Kidney (MDCK) cells were treated with CsA alone and CsA plus EF (25 and 50 μg/ml). The mechanism of attenuation of apoptosis and ROS by EF was studied using various experiments. Results: EF protected the MDCK cells from CsA-induced apoptosis and cell cycle arrest at G0/G1and sub G0/G1. The protective effect was further confirmed by cytochrome c translocation into cytoplasm and caspase 3 release. In addition, CsA-induced ROS production was abolished by the antioxidant potential of EF, which could be attributed to the polyphenol content. The decrease in ROS generation was confirmed by DCF-DA staining, LPO, and nitric oxide release in the MDCK cells. Moreover, the accumulation of collagen was decreased in EF-treated groups and increased the survivability of cells. The gas chromatography–mass spectrometry analysis revealed the presence of alkaloid and phenolic compounds. To the best of our knowledge, this is the first report showing the presence of alkaloid Carnegine and N-Benzyl-2-phenethylamine in this plant. Conclusion: Hence, it could be justified that EF-protected MDCK cells against CsA stimulated renal cell damage by attenuating apoptosis and ROS generation. Further isolation of compound has to be carried out to explore the effectiveness of EF as nephroprotective drug. Abbreviations used: CsA: Cyclosporine A; EF: Ethyl acetate fraction; CE: Crude extract; HF: Hexane fraction; CF: Chloroform fraction; RF: Residual fraction.
  2,166 122 -
Effect of aqueous/methanolic extract of Ocimum sanctum (OciBest) on the Male and Female reproductive performance of wistar rats
P Raina, B Bharathi, A Senthilkumar, S Sagar, CV Chandrasekaran, D Prabhu, M Deepak, A Amit, R Kaul-Ghanekar
April-June 2018, 14(55):256-263
DOI:10.4103/pm.pm_562_17  
Background: Ocimum sanctum (OSE) has been used in the Indian system of traditional medicine (Ayurveda) for the treatment and prevention of various diseases. However, the contradictory reports regarding the reproductive safety of OSE prompted us to re-verify its effect on the reproductive system by following internationally accepted Organization for Economic Cooperation and Development guideline 415. Objective: To study the effect of aqueous/methanolic extract of OSE (OciBest) on the male and female reproductive performance of Wistar rats. Materials and Methods: Rats were orally gavaged with OciBest at dose levels of 0, 250, 500, and 1000 mg/kg. Males were administered with OciBest for 12 weeks before and during the mating period. On the other hand, females received OciBest for 2 weeks before mating until the end of the lactation period. Results: OciBest at 1000 mg/kg did not induce any adverse effects on the reproductive performance of male and female rats. All the treated parent animals survived until the end of the study period with no major signs of clinical toxicity. The body weights, food consumption, male and female fertility indices, organ weights, as well as gross pathological and histopathology observations in parent animals, did not reveal any treatment-related adverse effects. Moreover, in comparison to the control groups, OciBest did not induce any treatment-related adverse effects to the offsprings. Conclusion: Thus, no-observed adverse effect level was found up to 1000 mg/kg dose, suggesting the safety of OciBest for reproductive system. Abbreviations used: OECD: Organization for Economic Cooperation and Development; NOAEL: No-observed adverse effect level; CPCSEA: Committee for the Purpose of Control and Supervision of Experiments on Animals; OA: Oleanolic acid; UA: Ursolic acid.
  2,126 160 -
Biological effects of Salvia Officinalis leaf extract on murine myeloma cells
Elisa Ovidi, Doriana Triggiani, Maria Valeri, Fabio Mastrogiovanni, Laura Salvini, Claudia Bonechi, Anna Rita Taddei, Valentina Laghezza Masci, Antonio Tiezzi
April-June 2018, 14(55):208-212
DOI:10.4103/pm.pm_401_17  
Background: In the Mediterranean region, Salvia officinalis plant is widely used and routinely added to food for a traditional culinary utilization, and moreover, from long time, it is recognized to possess pharmacological activities. Objectives: In the present study, we investigated the biological properties of S. officinalis leaf ethanol extracts and a thin layer chromatography-isolated spot on murine myeloma cells. Materials and Methods: 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide tests were carried out to determinate the EC50 of both the leaf extracts and the isolated spot; the isolated spot was also investigated by liquid chromatography–mass spectrometry (LC-MS) and nuclear magnetic resonance (NMR). Electron microscopy and immunofluorescence were used for morphological studies on treated P3X murine myeloma cells. Results: LC-MS and NMR techniques identified methyl carnosate, a methyl derivative of carnosic acid, as the main component of spot B. Moreover, scanning electron microscopy, transmission electron microscopy, and immunofluorescence investigations carried out on murine myeloma cells treated for 20 h with EC50 values of spot B revealed some changes both in the cellular morphology and in the microtubular array. Conclusions: The present studies indicate that S. officinalis extracts have biological effects on murine myeloma cells and identify methyl carnosate as an interesting molecule for further investigations on human cell lines and possibly on cancer prevention. Abbreviations used: EC50: Effective concentration 50, LC-MS: Liquid chromatography–mass spectrometry, NMR: Nuclear magnetic resonance, TLC: Thyn layer chromatography, MTT: 3-[4,5-dimethylthiazol-2-yl]-2, 5-diphenyltetrazolium bromide, RF: Retention factor, MS/Ms: Tandem mass spectrometry, dqf-COSY: double quantum filter COSY
  2,122 159 -
Green synthesis and antioxidant analysis of In vivo Leaf and In vitro Callus of Tephrosia villosa
V Ranjitha, K Kalimuthu, V Chinnadurai, Y Sharmila Juliet, M Saraswathy
April-June 2018, 14(55):147-153
DOI:10.4103/pm.pm_64_17  
Background: Tephrosia villosa (Leguminosae) is an important medicinal plant widely used in traditional Indian medicine. To the best of our knowledge, there is no report on this field. Objective: The aim of this study was to synthesize silver nanoparticles (SNPs) using T. villosa in vivo leaf and in vitro callus extract and characterization of SNPs. Materials and Methods: The synthesized SNPs were characterized by ultraviolet (UV) visible spectrum, Fourier transform infrared (FT-IR), X-ray diffraction (XRD), scanning electron microscopic (SEM), and Energy-dispersive X-ray spectroscopy (EDX). The in vitro antioxidant activities of the synthesized nanoparticles were studied by 1, 1-diphenyl-2-picryl-hydrazyl radical scavenging activity, hydrogen peroxide assay, nitric oxide radical scavenging activity, and hydroxyl radical scavenging activity. Results: The in vivo leaf SNPs have absorbance peak at 740 nm, and callus SNPs have absorbance peak at 305 nm for reaction at room temperature. The FTIR results also showed interaction between the plant extract and Ag-NPs due to the similarity in the peak patterns. The four and nine diffraction peaks were obtained from leaf and callus SNPs, respectively, through XRD studies. The obtained SEM image confirms the presence of polymorphic nanoparticles such as spherical, ellipsoidal, and some irregular shaped. The EDX results showed that Ag-NPs display an absorption peak at 3 keV, indicating the presence of the element silver. Conclusion: It is confirmed that both the in vivo leaf and in vitro callus SNPs revealed high level of antioxidant activity in in vitro system. Abbreviations used: AgNO3: Silver Nitrate; mg: Mille gram; ml: Mille litter; mM: Micro molar; μg: Micro gram.
  2,162 110 -
Pharmacokinetic study on piplartine and piperine after oral administration of Piper chaba root by liquid chromatography-mass spectrometry/mass spectrometry
PR Rao Vadaparthi, Kumar Katragunta, AK M. Pawar, Suresh Babu Katragadda, Ashok K Tiwari, Madhusudana Kuncha
April-June 2018, 14(55):161-166
DOI:10.4103/pm.pm_470_17  
Background: Piperaceae family are a well-known source of structurally diverse amides with the wide range of bioactivities such as cytotoxic, stomach aches, insect repellents, anti-inflammatory, insecticidal, and antifeedant activities. It has been reported that piplartine and piperine, alkaloid/amide compounds from Piper species, show antitumor activities. Objective: A rapid, sensitive liquid chromatography-tandem mass spectrometry method has been developed and validated for the determination of piplartine and piperine from Piper chaba extract. Materials and Methods: The two analytes, together with internal standard (IS, trichostachine), were separated on a Waters Acquity ethylene bridged hybrid C18(2.1 mm × 100 mm, 1.9 μ) column using a mobile phase of acetonitrile with 0.1% formic acid and water with 0.1% formic acid (70:30, v/v) with isocratic elution. The detection was performed using the positive ion electrospray ionization in multiple reaction monitoring mode with transitions at m/z 318→221 for piplartine, m/z 286→201 for piperine, and m/z 272→201 for the IS. Results: The calibration curves were both linear (r2 > 0.995) over a concentration range of 1.0–2000 ng/mL; the lower limit of detection quantification was 1.0 ng/mL for both piplartine and piperine. The intra-day and inter-day precisions (relative standard deviation %) were <10.9%, and recoveries ranged from 90.3% to 103.0%. Conclusions: The analytes were proven stable in the short-term, long-term, and after three freeze-thaw cycles. The method was successfully applied to pharmacokinetic studies of piplartine and piperine in rats after oral administration of P. chaba extract. Abbreviations used: AUC: Area under the curve; BEH: Ethylene bridged hybrid; CDER: Centre for drug evaluation and research; CID: Collision-induced dissociation; Cmax: Maximum concentration; CTO: Column Temperature Oven; DGU: Degassing Unit; ESI: Electrospray ionization; eV: Electron volt; FCV: Flow control valve; HPLC: High-pressure liquid chromatography; HPTLC: High performance thin layer chromatography; IS: Internal standard; LLOQ: Lower limit of quantitation; LC: Liquid chromatography; LC-MS: Liquid chromatography-Mass Spectrometry; LC-MS/MS: Liquid chromatography-Mass Spectrometry/Mass Spectrometry; LC-HRMS: Liquid chromatography-High resolution mass Spectrometry; LC-NMR-MS: Liquid chromatography-Nuclear magnetic resonance-Mass Spectrometry; MRM: Multiple reaction monitoring; MC: Methyl cellulose; N2: Nitrogen; RSD: Relative standard deviation; RE: Relative error; r2: Regression coefficient; t1/2: Half-life; Tmax: Time to maximum effect; QC: Quality control; UFLC: Ultrafast liquid chromatography; UPLC-qTOF-MS: Ultra pressure liquid chromatography-Time of flight-Mass spectrometry; USFDA: United states Food and Drug Administration
  2,085 158 -
Taxifolin possesses anti-cancer activity on the 7,12-Dimethylbenz(a)anthracene-Induced breast cancer in the sprague dawley rats by remodeling nuclear factor Erythroid 2- Kelch-Like ECH-Associated Protein 1-Heme Oxygenase 1 and anti-oxidant pathways
Md. Wasimul Haque, Mohd. Usman Mohd Siddique, Pritha Bose, Shakti Prasad Pattanayak
April-June 2018, 14(55):110-117
DOI:10.4103/pm.pm_601_17  
Background: In mammary cancer, alterations in various gene expressions and signaling pathways occurs due to the secondary effects of oxidative stress that facilitates cancer by causing genomic instability and mutagenic alterations. Several phenolic compounds are active against various malignancies. Taxifolin (TAX) exhibits diverse bioactivity profile that also contributes toward its anticancer efficacy. Objective: The present study has been designed for estimation of the anticancer potential of TAX on 7,12-dimethylbenz(a)anthracene (DMBA)-induced breast cancer in Sprague Dawley (SD) rats. Materials and Methods: Molecular docking analysis of Kelch-like ECH-associated protein 1 (Keap-1) and heme oxygenase-1 (HO-1) was carried out using Maestro tool to rationalize the activity of TAX based on their binding potential. This was followed by DMBA administration in air pouch to induce mammary cancer in female SD rats (50–55 days old). After 90 days of cancer induction, the chemotherapeutic potential of TAX was evaluated by the administration of TAX at doses 10, 20, and 40 mg/kg/day. Besides this, the effect of TAX on Keap-1-nuclear factor erythroid-2 (Nrf-2) pathway associated with HO-1 and NADPH:quinoneoxidoreductase 1 (NQO1) expressions and their effect on the anti-oxidative and anti-proliferative activity was also evaluated through immunofluorescence analysis, real-time quantitative polymerase chain reaction, and biochemical estimations. Results: TAX revealed protective effect against lipid peroxidation, enzymatic (superoxide dismutase [SOD], manganese-containing SOD, copper- and zinc-containing SOD, catalase, and glutathione peroxidase), and nonenzymatic (reduced glutathione, α-tocopherol, and ascorbic acid) anti-oxidative markers in serum, liver, kidney, and breast tissue of both control and experimental groups. The study revealed upregulation of protective Nrf-2, HO-1, and NQO1 expressions with consequent suppression in Keap-1 mRNA expression. Conclusion: This study revealed the potency of TAX in the inhibition of mammary carcinogenesis through Nrf-2-Keap-1-HO-1 and antioxidant pathway. Abbreviations used: RT-qPCR: Real-time quantitative polymerase chain reaction; ODC: Ornithin decarboxylase; HO-1; Heme oxygenase-1; PAHs: Polyaromatic hydrocarbons; TBARS: Thiobarbituric acid reactive species; NQO1: NADPH:quinoneoxidoreductase 1; Keap-1: Kelch-like ECH associated protein 1; Nrf2: Nuclear factor erythroid 2; SD: Sprague Dawley.
  2,013 143 -
High performance thin layer chromatography-automated multiple development bioautography of phytoconstituents and quantification of stigmasterol in Monochoria vaginalis and Monochoria hastata with antioxidant potential
Narayanan Kasthuri Bai, Kaliappan Ilango
April-June 2018, 14(55):45-51
DOI:10.4103/pm.pm_71_17  
Context: Monochoria vaginalis and Monochoria hastata belonging to Pontederiaceae are edible aquatic herbs commonly used by ethnic communities of India for treating various afflictions. Objective: The main objective of the study is to propose a simple and a rapid bioautographic fingerprinting profile using high-performance thin-layer chromatography (HPTLC) with gradient elution for screening the phytochemicals for the antioxidant property. Quantification of stigmasterol is carried in selected parts of Monochoria genus. Materials and Methods: Stigmasterol content of the three different parts of the species were quantified and validated by HPLTC. Fingerprint analysis was carried out using HPTLC-automated multiple development 2 (HPTLC-AMD2)-based gradient elution technique. Bioautography was done using 1-diphenyl-2-picrylhydrazyl (DPPH) solution to check the antioxidant property. Phytochemicals such as phenols, flavonoids, sterols, and saponins were estimated using 96-well plate and antioxidant potential were confirmed by DPPH, hydroxyl and nitric oxide scavenging activity. Results: The developed HPTLC-AMD2 method showed clear separations resulting in sharp, intense peaks. Phytoconstituents were determined for the first time in the Monochoria species. The results indicate that both species are rich in various bioactive contents with potent antioxidant potential. Stigmasterol was found to be present in all the selected parts with varying concentration. Conclusion: This study reveals that Monochoria species are rich in phytoconstituents with potent antioxidant activity and the developed method is efficient, simple, rapid, and reliable for separating the phytoconstituents of Monochoria providing a passport data of extracts. A positive, significant linear relationship between antioxidant activity and total phenol content and total flavonoid content showed that phenolic compounds and flavonoids were the dominant antioxidant components present in the extracts. Abbreviations used: HPTLC-AMD: High-performance Thin-layer Chromatography-Automated Multiple Development; MHL-Monochoria hastata leaf, MHS-Monochoria hastata stem; MHR-Monochoria hastata Rootstock; MVL-Monochoria vaginalis leaf; MVS-Monochoria vaginalis stem; MVR-Monochoria vaginalis Rootstock. TPC-Total phenol content; TFC-Total flavonoid content; TSC-Total sterol content; TSAC-Total saponin content; TTC-Total triterpenoid content.
  2,010 124 -
Media standardization for enhanced production of bacoside of Bacopa monnieri In situ condition
Pratibha Anil Chaturvedi, Lal Hingorani
April-June 2018, 14(55):32-35
DOI:10.4103/pm.pm_328_17  
Background: Secondary metabolites are known to not only play a major role in the adaptation of plants to their environment but also represent an important source of active pharmaceuticals Accumulation of such metabolites often occurs in plants subjected to stresses including various elicitors or signal molecules. The present study describes the enhancement of active principal bacoside of Bacopa monnieri by using stress and precursor compound. Materials and Methods: The study was done in situ condition. With incorporation of precursor compounds (phenylalanine and tyrosine [Ty]) and stress-inducing compounds (ZnCl2, CoCl2, and CuSo4). Results: The maximum content of bacoside recorded in 7-day-old plant treated with 25 μM CoCl2 (5.313%) separately followed by 15-day-old plants treated with 60 mg/100 ml Ty (5.15%) and 15-day-old plants treated with 50 μM of CoCl2 (5.15%). Conclusion: Enhancement of bacoside in B. monnieri was observed more than 2% than in vivo condition by giving CoCl2stress in situ condition. This kind of experiment is not well documented till date. Abbreviations used: BM :Bacopa monerrie; Ty: Tyrosine; Phy: Phenylalanine
  1,954 122 -
Quinic acid attenuates oral cancer cell proliferation by downregulating cyclin D1 Expression and Akt signaling
Anjana Singh, Shyam Singh Chauhan, Vishwas Tripathi
April-June 2018, 14(55):14-19
DOI:10.4103/pm.pm_36_18  
Background: Quinic acid (QA), a natural compound found in fruits, for example in apple, berries, and coffee beans, is a neutraceutical chiral compound used to synthesize new pharmaceutically important chemical compounds. Platinum-based drugs such as cisplatin are widely used in the therapy, including oral cancer, one of the most prevalent cancer. Due to their severe side effects and acquired resistance to the treatment, phytotherapies are being explored to find out strong anticancer compounds that can alone and synergistically prevent and cure cancer without side effects. Objective: This study aimed to find out the mechanism behind the anticancer property of QA, a natural compound alone and synergistically with cisplatin in oral cancer cells (squamous cell carcinoma-4 [SCC-4]). Materials and Methods: Oral cancer cells were treated with QA alone and synergistically with cisplatin. MTT assay was performed to investigate the cytotoxic effect of QA and QA + cisplatin combination. Apoptotic effects of QA and QA + cisplatin combination were analyzed using DAPI and real-time-polymerase chain reaction for the expression of apoptotic marker genes. Effect of QA and QA + cisplatin combination was assessed by flow cytometry and immunoblotting technique. Results: Our result shows that QA promotes apoptosis in oral cancer cells by downregulating the expression of anti-apoptotic genes and attenuating the expression of cyclin D1 and Akt signaling pathway. Moreover, QA shows synergistic effect with cisplatin in these cells. Conclusion: QA inhibits cell proliferation and promotes apoptosis in oral cancer cells (SCC-4) alone and with cisplatin. Abbreviations used: QA: Quinic acid; CDK: Cyclin-dependent kinase; SCC-4: Squamous cell carcinoma-4; FBS: Fetal bovine serum.
  1,900 139 -
High-performance liquid chromatography identification of gastroprotective and antioxidant effects of purified fractions A-E from the stem of Coscinium fenestratum
Hui Yang, Hong-Bin Zhai, Wei-Min Wang, Okechukwu Patrick Nwabueze
April-June 2018, 14(55):78-83
DOI:10.4103/pm.pm_267_17  
Objective: Coscinium fenestratum belongs to the family of Menispermaceae. It is a woody plant that originated from Indomalaya regions such as southern India, Sri Lanka, Thailand, Vietnam, Cambodia, and West Malaysia. C. fenestratum has been reported to possess various pharmacological actions such as antioxidant, antiproliferative, antidiabetic, and antibacterial activities. In addition, this medicinal plant has also been used to treat gastrointestinal disorders, including ulcer. The purified fractions A-E of dichloromethane (DCM) extract obtained from the stem of C. fenestratum were investigated for antioxidant activity and its ability to protect the gastric mucosa against hydrochloric acid (HCl)/ethanol, nonsteroidal anti-inflammatory drugs (NSAIDs), and stress-induced ulcer. The bioactive compounds were identified using high-performance liquid chromatography (HPLC) and simple biochemical screening. Methods: Air-dried stem extract was soaked in DCM to obtain crude extract which was purified using column chromatography. HPLC and basic biochemical screening were used to identify the bioactive compound present. HCl/ethanol induced, NSAIDs, and stress of animal were used to induce gastric ulcer. The rats were orally administered with purified A-E extract for gastroprotective and antioxidant activity. Results: The oral administration of fractions A-E exhibited gastroprotective activity by reducing the ulcerative index induced by HCl/ethanol, NSAIDs, and stress compared to saline. The extract showed high antioxidant effect, especially fraction E. The HPLC and phytochemical analysis of fraction E showed the presence of palmatine and flavonoid. Conclusion: The extract showed a very strong gastroprotective and antioxidant activities and the highest activity was observed by fraction E. The activity may be because of the presence of palmatine and flavonoid. Abbreviations Used: DCM: Dichloromethane, HCl: Hydrochloric acid, HPLC: High-performance liquid chromatography, NSAIDs: Nonsteroidal anti-inflammatory drugs, EtOH: Ethyl acetate, MeOH: Methanol, NaCl: Sodium chloride, FCR: Folin–ciocalteu reagent, DPPH: 2, 2-diphenyl-1-picrylhydrazyl, TLC: Thin-layer chromatography, IMR: Institute of Medical Research, TPC: Total phenolic compounds, UPM: University Putra Malaysia, Rf: Retardation factor, LD50: Lethal dose.
  1,887 124 -
EDITORIAL
DNA barcode testing in authentication of botanical raw material coming of age
DB A. Narayana, T Sudhakar Johnson
April-June 2018, 14(55):1-2
DOI:10.4103/pm.pm_249_18  
  1,832 123 -
ORIGINAL ARTICLES
Genistein-attenuated hepatic steatosis and inflammation in nonalcoholic steatohepatitis with bilateral ovariectomized rats
Sudaporn Pummoung, Duangporn Werawatganon, Naruemon Klaikeaw, Prasong Siriviriyakul
April-June 2018, 14(55):20-24
DOI:10.4103/pm.pm_603_17  
Objective: The objective was to investigate the anti-lipidemic and anti-inflammatory effects of genistein on rats with estrogen deficiency and nonalcoholic steatohepatitis (NASH). Materials and Methods: Sprague-Dawley female rats (n = 48) were randomly divided into ovariectomized (OVX) and non-OVX groups, then again divided into three subgroups as follows: controls, rats fed with high-fat high-fructose (HFHF) diet (NASH group), and rats fed with HFHF diet plus daily 16 mg/kg genistein (genistein group). Liver tissues were used for histology, liver tissues were used for histology and measured of hepatic free fatty acid (FFA) by colorimeter, and nuclear factor kappa B (NFκB) expression by immunohistochemistry. Serum tumor necrosis factor-α (TNF-α) was evaluated by enzyme-linked immunosorbent assay. Results: NASH group had increased serum TNF-α (171.62 ± 22.34 vs. 58.47 ± 14.83 pg/mL), %NFκB-positive cells (53.94 ± 11.89 vs. 13.73 ± 3.40), and hepatic FFA (9.07 ± 2.27 vs. 3.62 ± 0.77 nmol/mg tissue) when compared with control (P < 0.01). The most severe hepatic fat accumulation and inflammation was found in OVX with NASH group. Genistein treatment decreased serum TNF-α compared with NASH groups in both non-OVX and OVX groups (105.84 ± 29.77 vs. 171.62 ± 22.34 pg/mL and 73.07 ± 19.31 vs. 124.12 ± 16.04 pg/mL, respectively) (P < 0.01). Genistein reduced %NFκB-positive cells in NASH rats (31.84 ± 10.60 vs. 53.94 ± 11.89) and decreased hepatic FFA levels in OVX with NASH rats (6.50 ± 0.60 vs. 13.11 ± 1.65 nmol/mg tissue) when compared with NASH group, respectively (P < 0.01). Conclusion: Estrogen deficiency is the contributing factor that worsens NASH. Genistein attenuated hepatic fat accumulation and inflammation. Moreover, genistein demonstrated to be more effective in estrogen deficiency with NASH than ovary-intact rats. Abbreviations used: NAFLD: Nonalcoholic fatty liver disease; NASH: Nonalcoholic steatohepatitis; non-OVX: Nonovariectomized; OVX: ovariectomized; HFHF: High-fat high-fructose; FFA: Free fatty acid; NFκB: Nuclear factor kappa B; TNF-α: Tumor necrosis factor-alpha; IL-6: Interleukin-6; PPARγ: Peroxisome proliferator-activated receptor gamma; ERα: Estrogen receptor alpha; ERβ: Estrogen receptor beta; LDL: Low-density lipoprotein; TBARS: Thiobarbituric acid-reactive substances; ELISA: Enzyme-linked immunosorbent assay; DMSO: Dimethyl sulfoxide; DAB: Diaminobenzidine.
  1,809 138 -
5,7-Dihydroxy-4-Methoxyflavone a bioactive flavonoid delays amyloid beta-induced paralysis and attenuates oxidative stress in transgenic Caenorhabditis elegans
Jyotsna Asthana, BN Mishra, Rakesh Pandey
April-June 2018, 14(55):57-64
DOI:10.4103/pm.pm_290_17  
Background: The various herbal remedies have been used in Ayurveda which symbolizes traditional medicine system since ancient times. The increasing popularity of herbal medicines has prompted us toward the development of natural therapeutics for preventing neurodegenerative disease such as Alzheimer's disease (AD) in living organisms. This study focused on a flavonoid compound 5,7-dihydroxy-4-methoxyflavone (DMF) also known as acacetin which is a major constituent of Premna odorata (L.) plant. This bioactive flavonoid exhibits several medicinal properties such as antimicrobial, anti-inflammatory, antioxidant, and anti-carcinogenic. Objective: The aim is to determine the protective effects of DMF against amyloid beta (Aβ)-induced toxicity and oxidative stress in transgenic Caenorhabditis elegans model of AD. Materials and Methods: The therapeutic potential of DMF treatments (5, 25, and 50 μM) was investigated to counteract Aβ paralysis and oxidative stress through paralysis assay, reactive oxygen species (ROS) detection, protein carbonylation, aldicarb assay, and mRNA quantification using transgenic C. elegans model of AD. Results: The present study reports that DMF effectively delayed Aβ-induced paralysis, attenuated ROS level reduced protein carbonylation and conferred aldicarb resistance. In addition, DMF was also found to up-regulate the expression of stress modulating (sod-1, sod-2, sod-3, ctl-1, hsp-16.2, and gst-4), acetylcholine transporter(unc-17), regulator of nicotinic acetylcholine receptor (unc-50), and choline acetyltransferase (cha-1) related genes. Conclusion: These findings suggest DMF may provide protection against Aβ toxicity and oxidative stress due to its antioxidant activity. Therefore, the bioactive flavonoid DMF may provide invaluable medicinal and health benefits which can delay the onset of age-related diseases. Abbreviations used: DMF: 5, 7-dihydroxy-4-methoxyflavone; ROS: Reactive oxygen species; C. elegans: Caenorhabditis elegans; AD: Alzheimer's disease; Aβ: Amyloid beta; DMSO: Dimethyl sulphoxide; NGM: Nematode growth media; ACh: Acetylcholine; Ald: Aldicarb; APP: Amyloid precursor protein.
  1,798 125 -
6-Gingerol prevents free transition metal Ion [Fe (Ii)] Induced free radicals mediated alterations by In vitro and Ndv growth in chicken eggs by In ovo
Ganjikunta Venkata Subbaiah, Kesireddy Sathyavelu Reddy, Yagani JayavardhanaRao, Bhasha Shanmugam, Sahukari Ravi, Kondeti. Ramudu Shanmugam, Golla Narasimha
April-June 2018, 14(55):167-174
DOI:10.4103/pm.pm_387_17  
Background: The study of free radicals and their reactions is implicated in the pathogenesis of various human diseases. Therefore, antioxidants treatment plays a pivotal role in preventing the free radical-induced diseases. Objective: The objectives of this study were to examine the protective role of 6-gingerol from free transition metal ion-induced alterations by in vitro and Newcastle disease virus (NDV) in chicken eggs by in ovo. Materials and Methods: In this study, lipid peroxidation, DNA sugar oxidation, chelation of ferrous ions, DNA fragmentation, Cytochrome c-oxidation, ferric ion reducing activities and DPPH, superoxide and hydroxyl radicals scavenging activity by ESR studies were examined with 6-gingerol by in vitro. Furthermore, antiviral activity was screened with 6-gingerol by in ovo. Results: In vitro results showed that 254 μM/ml of 6-gingerol firmly inhibited erythrocyte membrane lipid peroxidation by 85.28% and DNA sugar oxidation by 91.26% when compared with butylated hydroxyanisole 74.42% (554 μM/ml) and 78.30% (824 μM/ml), respectively. Whereas, chelating activity of 6-gingerol was 80.2% which nearly similar to the EDTA activity (50 μg/ml) shown 83.93%. In addition, 6-gingerol (254μM/ml) scavenged the superoxide, hydroxyl, DPPH radicals by 84.96%, 90.45%, and 94.63% respectively and the same was strongly supported with ESR studies. Further, 254 μM/ml of 6-gingerol inhibited the DNA fragmentation, Cytochrome c-oxidation and reducing the ferric ion by in vitro and antiviral activity shown on NDV at 100 μg/ml. Conclusion: This study could enlighten that the 6-gingerol has a good metal sequester property and it could prevent the free radicals by in vitro and NDV growth in chicken eggs by in ovo. Abbreviations used: NDV: Newcastle Disease Virus; ESR: Electron Spin resonance; SOD: Super oxide Dismutase; HA: Hemagglutination.
  1,825 96 -
Intercomparative investigation of the total phenols, total flavonoids, In vitro and In vivo antioxidant activities of capparis Cartilaginea (Decne.) maire and weiller and Capparis Ovata Desf. from Jordan
Mahmoud A Al-Qudah, Safwan M Obeidat, Riyadh Muhaidat, Bahaa Al-Trad, Hala I Al-Jaber, Jamil N Lahham
April-June 2018, 14(55):154-160
DOI:10.4103/pm.pm_356_17  
Background: Plant-derived antioxidants have been products of choice in therapeutic formulations for their potent free radical scavenging activity and mitigating and curing illnesses and diseases associated with oxidative stress. Capparis L. includes species with unprecedented nutritional and medicinal values, and hence offers a diverse pool of phytochemicals implicated in antioxidant activity. Objective: The current study was designed to assess the chemical variation, total phenols content (TPC), total flavonoids content (TFC), in vitro and in vivo antioxidant activities of different extracts obtained from Capparis cartilaginea and Capparis ovata from Jordan. Materials and Methods: The ethanolic extract from each species was partitioned in different solvents and the TPC, TFC, and antioxidant activity for each extract was examined. The in vitro antioxidant activities were tested by the 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2'-Azino–bis (3-ethylbenzoline-6-sulfonic acid diammonium salt (ABTS) radical scavenging methods in addition to the metal ion chelating effect (MCE) and hydroxyl radical assay methods. The butanol extract was investigated for its in vivo effect on the activities of serum superoxide dismutase (SOD) and glutathione peroxidase (GPX) in mice (100 mg/kg, intraperitoneally for 12 days). Results: The butanol fraction of both plants had a dose-dependent in vitro antioxidant activity. The treatment of mice with the butanol extract of both species for 12 days significantly increased the activities of serum SOD and GPX. Principal component analysis indicated that DPPH, TFC, hydroxyl radical had major variability in the antioxidant activities of the two investigated Capparis species. Results of this study underscores enrichment of the two studied Capparis species with phenolics and flavonoids that could account for much of their antioxidant activities. Abbreviations used: C. cartilaginea: Capparis cartilaginea ; C. ovata: Capparis ovata; TPC: Total phenols content; TFC: Total flavonoids content; DPPH: 2,2-diphenyl-1-picrylhydrazyl; ABTS: 2,2'-azino-bis-3-ethylbenzothiazoline-6-sulphonic acid; MC: Metal Chelating effect; Hydroxy: Hydroxyl radical assay; SOD: Serum superoxide dismutase; GPX: Glutathione Peroxidase ;PC: Principal Component.
  1,785 120 -
Rubus sanctus Schreb. root extract alters the MicroRNA expression and inhibits tumor activities of colorectal cancer cell lines
Adem Alemdar, Berrin Tunca, Hulusi Malyer, Saliha Sahin
April-June 2018, 14(55):92-101
DOI:10.4103/pm.pm_357_17  
Background: Colorectal cancer (CRC) is one of the most common cancers in the world. Although surgical and screening techniques have vastly improved in the last 30 years, chemotherapeutics have not advanced sufficiently for successful treatment. Objective: The aim of this study is to investigate the microRNA (miRNA) expression changes and anticancer agent potential of Rubus Sanctus Schreb. root extract (RRE) on LoVo and HT-29 colorectal adenocarcinoma cell lines. Materials and Methods: LoVo and HT-29 CRC cell lines treated with different concentrations of RRE to find growth inhibitory effect with WST-1 assay. Fifty percent growth inhibition and 25% growth inhibition concentrations further evaluated with annexin V, total caspase, cell cycle, and migration assays. Real-time polymerase chain reaction was used to investigate the expression differences in miRNA after extract treatment. Results: Cell proliferation was reduced 77.98% in HT-29 cells after RRE treatment (P < 0.05). In the cell invasion analysis, RRE reduced invasion in both cell lines up to 75.56% (P < 0.05). In addition, RRE induced apoptosis in up to 98% of a cell population (P < 0.05). Similarly, pan-caspase activity increased to 97.6% and 87.2% in LoVo and HT-29 cell lines, respectively (P < 0.0001). After extract treatment, among the nine miRNAs evaluated, only miR-140 expression was significantly increased in both cell lines after RRE treatment (P < 0.05). Conclusion: Our data show for the first time that RRE has the capability to inhibit CRC cell proliferation and invasion and alter epigenetic mechanisms. Although further studies should be conducted on this topic, RRE is thought to be a potential candidate for the future studies regarding new therapy options. Abbreviation used: RRE: Rubus Sanctus Schreb. root extract; CRC: Colorectal cancer; GI50, GI25: 50% and 25% growth inhibition; miRNA: MicroRNA; MGMT: O-6-Methylguanine-DNA methyltransferase; UTR: Untranslated region; Akt: Protein kinase B; PI3K: Phosphoinositide 3-kinase; NF-κB: Nuclear factor kappaB; iNOS: Inducible nitric oxide synthase; COX-2: cyclooxygenase 2; TNFα: tumor necrosis factor alpha; IL-6: Interleukin 6; DMEM: Dulbecco's Modified Eagle's medium; RPMI-1640: Roswell Park Memorial Institute 1640 medium; HPLC: High-performance liquid chromatography; UV-Vis: Ultraviolet-visible; FBS: Fetal bovine serum; OD: Optical density; RFU: Relative fluorescence unit; PE: Phycoerythrin; 7-AAD: 7-Aminoactinomycin D; PBS: Phosphate-buffered saline; EGFR: Epidermal growth factor receptor; 5-FU: Fluorouracil; MSI: Microsatellite instability; NSCLC: Nonsmall cell lung cancer; TGF-β: Transforming growth factor beta; EMT: Epithelial–mesenchymal transition
  1,715 123 -
Simultaneous quantification of Precocene I and Precocene II through high-performance thin layer chromatography validated method in Ageratum conyzoides L. germplasms from western himalayas
Bhanu Kumar, Ankita Misra, Ajay Kumar Singh Rawat, Yashwant Singh Rawat, Sharad Srivastava
April-June 2018, 14(55):141-146
DOI:10.4103/pm.pm_411_17  
Background: Ageratum conyzoides L. is a traditionally used herb for various ethnoveterinary purposes. There are no earlier reports on simultaneous high-performance thin layer chromatography (HPTLC) quantification of bioactive markers. Objective: To develop a sensitive, robust, and replicable HPTLC method for simultaneous estimation of precocene I (PI) and precocene II (PII) in A. conyzoides L. collected from the Western Himalaya region of India, with an aim to understand the level of chemotypic differences arising in the intraspecific population of this ethnobotanically important plant. Materials and Methods: A sensitive HPTLC method was developed to resolve PI and PII using toluene: ethyl acetate (9.8:0.2 v/v) as mobile phase. The method was validated for selectivity, specificity, linearity, and precision as per International Conference on Harmonization guidelines. Results: Good linearity was achieved over a five-point concentration range with a correlation coefficient of 0.986 and 0.988 for PI and PII, respectively. The PI content varies in the range of 0.0016% (NAC-77) to 0.0834% (NAC-82), whereas PII was reported to be present in the range of 0.016% (NAC-85) to 0.143% (NAC-91) on a dry weight basis. A principal component analysis biplot of samples based on the content of PI and PII identified four elite chemotypes, namely, NAC-81, NAC-82, NAC-85, and NAC-91. Conclusion: The study identifies superior germplasms for commercial prospection and develops a validated method that can be used for the quality control of herbal drug/formulation using A. conyzoides as an ingredient. Abbreviations used: HPTLC: High performance thin layer chromatography, PI: Precocene I, PII: Precocene II, ICH: International conference on harmonization, SD: Standard deviation, RSD: Relative standard deviation
  1,669 128 -
A novel compound Lup-20 (29)-ene-3α,6β-diol identified in petroleum ether extract of diospyros melanoxylon Roxb. leaves and to reveal its antidiabetic activity in rats
Anil Kumar Sharma, Tek Chand Sharma, Ruchi Singh, Pratibha Payal, Rajnish Gupta, Mahesh Chandra Sharma
April-June 2018, 14(55):245-248
DOI:10.4103/pm.pm_429_17  
Background: Diospyros melanoxylon Roxb. belongs to the family Ebenaceae and its leaves show diuretic, carminative, laxative, and styptic bioactivities. Objective: The objective of this study was to give a direction for future investigators to carry out research on phytochemistry and develop a potent antidiabetic agent. Materials and Methods: Petroleum ether extract of D. melanoxylon Roxb. leaves was chromatographed over silica gel column. Isolated compounds were elucidated on the basis of different spectroscopic techniques (mass spectrometry, infrared, 1H, 13C nuclear magnetic resonance, and distortionless enhancement by polarization transfer). Antidiabetic activity of lup-20 (29)-ene-3α,6 β-diol was evaluated in streptozotocin diabetic rats. Results: A novel compound, lup-20 (29)-ene-3α,6 β-diol (6), was identified along with 11 known natural compounds, namely, lupeol (1), ceryl alcohol (2), octacosanol-1 (3), hentriacontanol-1 (4), β-sitosterol (5), diospyrin (7), 3α-methoxydiospyrin (8), betulin (9), ursolic acid (10), betulinic acid (11), and oleanolic acid (12). Further administration of lup-20 (29)-ene-3α,6 β-diol significantly improved the diabetes-induced oxidative stress. Conclusion: Investigation of this novel compound plays an important role in the field of drug development. Abbreviations used: STZ: (Streptozotocin); LPO: (Lipid Peroxidation); SOD: (Superoxide Dismutase); CAT: (Catalase); GSH: (Reduced Glutathione).
  1,690 103 -
Effect of simvastatin on neuroinflammation in microglial cells via mitogen-activated protein kinase and nuclear factor κB pathways
HongYing Ma, DingAn Li, Thamaraiselvan Rengarajan, Kalaivani Manokaran
April-June 2018, 14(55):237-244
DOI:10.4103/pm.pm_418_17  
Background: Activated microglial cells are found in different sorts of the neurodegenerative process including Parkinson and Alzheimer. Suppressing the activated microglial cells developed as a novel procedure for the treatment of neuroinflammation-based neurodegeneration. Materials and Methods: We have investigated the effects of simvastatin on memory impairment and inflammatory cytokines expression induced by transient cerebral ischemia in cultured microglial cells. Results: The lipopolysaccharide (LPS)-activated microglial cells treated with simvastatin 3 μmol has decreased the inflammation which was indicated by the reduced levels of the nitric oxide (NO), tumor necrosis factor-α, interleukin-1 β, cyclooxygenase-2, and inducible NO synthase. Simvastatin also delayed the activation of atomic component nuclear factor-κB, p38 mitogen-activated protein kinase, and the reactive oxygen species in LPS-activated microglial cells. Moreover, simvastatin has provoked the outflow of heme oxygenase-1 in BV-2 microglial cells. Conclusions: The present study showed that the simvastatin antagonizes neuroinflammation and can be a potential restorative operator for treating neuroinflammatory ailments. Abbreviations used: LPS: Lipopolysaccharide, TNF-α: Tumor necrosis factor, NO: Nitric oxide, IL-1β: Interleukin, COX-2: Cyclooxygenase, iNOS: Inducible nitric oxide synthase, MAPK: Mitogen-activated protein kinase, HO-1: Heme oxygenase
  1,596 116 -
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