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   2019| April  | Volume 15 | Issue 62  
    Online since April 26, 2019

 
 
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ORIGINAL ARTICLES
In vivo and in vitro antidiabetic and antioxidant activity of spirulina
Patrick Nwabueze Okechukwu, Sophia Ogechi Ekeuku, Mridula Sharma, Chong Pei Nee, Hor Kuan Chan, Norazlina Mohamed, Gabriele Ruth Anisah Froemming
April 2019, 15(62):17-29
DOI:10.4103/pm.pm_431_18  
Objective: This study aims to evaluate the effect of spirulina, a biomass produced by cyanobacteria, on the level of plasma glucose, oxidative stress, and other biochemical parameters in diabetes in streptozocin (STZ) 50 mg/kg-induced diabetic-induced rat model. Materials and Methods: The in vitro antioxidant property of spirulina was assessed by measuring its ability to scavenge free radicals and reactive oxygen species (ROS) such as superoxide anion, nitric oxide, and hydroxyl and lipid peroxyl radicals. The inhibition of diabetic link enzymes alpha-glucosidase, alpha-amylase, and dipeptidyl peptidase-4 inhibitor (DPP-IV) were tested in vitro. Thirty female Sprague-Dawley rats weighing 150–250 g were divided into five groups: normal, diabetes (negative control), metformin in single dose of 300 mg/kg, spirulina in a single dose of 300 mg/kg and spirulina combined with metformin at dose of 150 mg/kg, and spirulina at dose of 150 mg/kg (spirulina + metformin 300 mg/kg) (n = 6). After an acclimation period of 2 weeks, diabetes was induced in the rats through STZ intraperitoneal injection. Spirulina (300 mg/kg) was dissolved in water and was administered orally for 12 weeks, and the rats' that plasma glucose level reached ≥11 mmol/L after 12 weeks treatment was selected for the study. After the treatment, the blood and liver were used for the evaluation of antioxidant enzyme activities, lipid, liver, kidney, and hematology profile. Results: Spirulina was able to reduce hyperglycemia-induced oxidative stress by reducing plasma glucose levels and scavenging or reducing the production of ROS and free radicals. It was also able to inhibit the activities of the alpha-glucosidase, alpha-amylase, and DPP-IV. With this, it significantly reduced the effect of STZ on the liver and kidney at the organ level and on antioxidant enzymes at the cellular level. Conclusion: Spirulina is able to reduce the lipid, liver, and kidney disease markers in STZ-induced rats and therefore is a potential supplement for diabetic patients. The antidiabetic effect of spirulina may be based on the antioxidant effect of the biomass as a whole, or it is based on specific bioactive components present in spirulina.
  1,025 3 -
Androgen Receptor (NR3C4) regulator potential of Ceratonia siliqua extract and its signaling pathways
Fehmi Kigili, Fatih Ozen, Tunc Catal, Belkıs Atasever Arslan
April 2019, 15(62):1-4
DOI:10.4103/pm.pm_588_18  
Aim/Background: Androgen receptor (AR/NR3C4) regulates growth, development, reproduction, metabolism, and homeostasis. Since it has multifunctional properties, disrupting of the functions can cause many diseases such as cancer and neurodegenerative diseases. Therefore, AR modulators and blockers are very important therapy of androgen-dependent diseases. Furthermore, mechanisms and signaling pathways of AR on the diseases are unclear. Ceratonia siliqua was shown to have preventative effects against digestive system disorders, diabetes mellitus, asthma-bronchitis, and oxidative stress in various studies. Materials and Methods: In the present study, its AR regulator potential was investigated in AR-deficient HEK293 cells. Results: While C. siliqua extract increased >2 folds of NR3C4 gene expression in the cells, it induced a decrease in AKT1 and kynureninase genes expression levels. On the other hand, it increased p38 gene expression, but it did not change FUS gene expression. These results support a regulating role on the receptor and also that the compounds specifically affect NR3C4 signaling pathways. Conclusion: Determination of the molecules and their various combinations with each other can contribute to discover new therapeutic agents for diseases dependent on AR signaling pathways.
  783 1 -
Antihyperglycemic activity of the leaves from annona diversifolia Safford. and farnesol on normal and alloxan-induced diabetic mice*
Fernando Calzada, Miguel Valdes, Normand Garcia-Hernandez, Claudia Velázquez, Elizabeth Barbosa, Celia Bustos-Brito, Leovigildo Quijano, Emmanuel Pina-Jimenez, Jesica Elena Mendieta-Wejebe
April 2019, 15(62):5-11
DOI:10.4103/pm.pm_582_18  
Background: Diabetes mellitus (DM) is a chronic disease characterized by high blood glucose levels resulting from insulin resistance or inadequate insulin secretion. In the world, DM is one of the most frequent non-contagious diseases that affect more than 371 million people. Objective: This study aimed to evaluate the antihyperglycemic properties of the ethanol extract, subsequent fractions, and farnesol obtained from the leaves of Annona diversifolia on alloxan-induced diabetic and normal mice. Materials and Methods: Bioassay-guided fractionation of the ethanol extract of the leaves of A. diversifolia (EELAd) was performed on alloxan-induced Type 2 diabetic and normoglycemic (NM) mice. Oral glucose tolerance test (OGTT), oral sucrose tolerance test (OSTT), and oral lactose tolerance test (OLTT) were performed in fast NM mice (FNM). Results: The EELAd, CHCl3 fraction, and farnesol induced a significant reduction of postprandial hyperglycemia in acute and subchronic tests using AITD mice. When EELAd, CHCl3 fraction, and farnesol were tested on NM in subchronic assays, these did not affect glycemic levels. In the case of acute test on NM, only CHCl3 fraction induced a hypoglycemic effect at 2 h after the treatment. OLTT and OSTT showed that the EELAd, CHCl3 fraction, and farnesol induced a significant reduction of hyperglycemia levels in FNM at 2 h after a lactose or sucrose load comparable to acarbose. In the case of OGTT was observed a significant reduction of hyperglycemia levels in FNM mice at 2 h after a glucose load comparable to canagliflozin. Conclusion: The EELAd and farnesol induced a significant reduction of postprandial hyperglycemia on AITD mice in acute and subchronic assays. Our results suggest that the control of postprandial hyperglycemia may be mediated by the regulation of absorption of glucose and inhibition of disaccharide digestion such as sucrose and lactose. Finally, the results explained the use of A. diversifolia in Mexican traditional medicine as an antihyperglycemic agent.
  781 0 -
In vitro antileishmanial activity of methanolic extracts for some selected medicinal plants
Ahmed Gomaa Gomaa Darwish, Mamdouh Nabil Samy, Sachiko Sugimoto, Katsuyoshi Matsunami, Hideaki Otsuka
April 2019, 15(62):34-37
DOI:10.4103/pm.pm_570_18  
Objective: The aim of this study is to evaluate the antileishmanial activity of selected medicinal plants; ten well-known medicinal plants cultivated and growing under African environmental conditions were studied. Materials and Methods: The methanolic extracts of these plants were screened for their antileishmanial activity against Leishmania major using 3-(4.5-dimethylthiazol-2yl)-2.5-diphenyltetrazolium bromide assay. Results: The methanol extract of Colchicum autumnale and Alpinia officinarum showed potent antileishmanial activity at inhibition% value of 98.29% ± 0.75% and 97.25 ± 1.63%, respectively, while Silybum marianum exhibited inhibition% value of 90.97% ± 1.13%, compared with the standard amphotericin B (89.31% ± 2.08%). Conclusion: Considering these results, medicinal plants from African environment could constitute a developer source for antileishmanial compounds.
  693 0 -
Physicochemical standardization of mucilage obtained from Althaea officinalis Linn – Root
Mazhar Husain, Abdul Wadud, Hamiduddin , Gulamuddin Sofi, Shaista Perveen, Khadeeja Abdul Hafeez
April 2019, 15(62):155-161
DOI:10.4103/pm.pm_506_18  
Introduction: Plant derived mucilage has been explored as a drug, pharmaceutical excipient, and in cosmetics. Several mucilage and mucilage-containing drugs are being utilized in Unani medicine. These are to be standardized for authentications owing to immense utilization. A mucilage-containing drug obtained from root of Althaea officinalis L. (AO) – family Malvaceae, an important Unani drug, has been subjected to physicochemical studies for standardization. Materials and Methods: Mucilage of roots of the drug was isolated by classical and reference method. The physicochemical method included determination of ash values, moisture content, viscosity, swelling index (SI), and pH value. Powder characterization study included bulk density, tapped density, Hausner's ratio, and angle of repose. For preliminary phytochemical analysis, qualitative tests for organic constituents and test for mucilage were carried out. Analytical methods, namely Fourier transform infra-red (FTIR) and high-performance thin-layer chromatography (HPTLC) were also applied. Results: The yield percentage taken by acetone method was 36.80 ± 1.25 whereas that of classical method was 42.93 ± 1.35. Values of pH, loss on drying, viscosity, and SI were 4.08 ± 0.032, 14.46 ± 0.13, 34.40 ± 0.61, and 334.36 ± 23.77, respectively. Data for ash value and powder characterization (Micromeritic Properties) were set in. Preliminary confirmative test confirmed that the isolated polysaccharide is mucilage. HPTLC fingerprinting of aqueous extract gave 6 and 4 peak at 254 nm, 4 and 5 peak at 366 nm and 5 and 6 peaks at 550 nm in mobile phase chloroform (90): methanol (10): acetic acid (2). FTIR data for the mucilage were also set in. Conclusion: Physicochemical standardization data/monograph for AO root mucilage were developed.
  676 0 -
14-Deoxy-11,12-Didehydroandrographolide: A novel compound isolated from Andrographis paniculata Nees. induces robust apoptosis in leukemic cells
Swadesh Sarkar, Priya K Gopal, Biswajit Chakraborty, Mausumi Paul, Chinmay Chowdhury, Santanu Paul
April 2019, 15(62):135-143
DOI:10.4103/pm.pm_466_18  
Objective: Andrographis paniculata is widely cultivated in South and Southeast Asian countries and popularly used in “Ayurveda” medicine. We attempted to investigate antileukemic activity of the biomolecules extracted from this plant and a probable mechanism of action. Materials and Methods: Biomolecules from methanolic extract were isolated using silica gel column chromatography and high-performance liquid chromatography. The structures were determined by liquid chromatography-mass spectrometry (LC-MS), 1H nuclear magnetic resonance (NMR), and 13C NMR. In vitro antiproliferative activity was measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The apoptotic efficacy of the most potent molecule was investigated by annexin V and propidium iodide (V/PI) staining and flow cytometry. Caspase activation, cell cycle distribution pattern, and nuclear morphology of the treated leukemic cells were also investigated. Results: From the methanolic extract, four biomolecules were isolated, namely 14-deoxy-11,12-didehydroandrographolide (1), andrographolide (2), neoandrographolide (3), and 14-deoxy-11,12-didehydroandrographiside (4). Results of MTT assay exhibited that out of four compounds, compound (1) showed the most potent activity against all the cell lines tested with the lowest IC50 values of 13 μM on U937 cells. Annexin V/PI staining revealed that the compound was able to induce apoptosis in concentration-dependent manner with IC50 value being 17.66 μM. Apoptotic induction was mediated through elevated activation of caspase-3 and caspase-9. Cell cycle analysis revealed that the compound (1) effectively increased the sub-G0-G1 population in the treated U937 cells (73.25% at 50 μM) in comparison to control set (3.12%). DAPI nuclear staining indicated that compound (1) increased the number of deformed nuclei and an increased level of apoptotic body formation in the treated cells.
  648 0 -
Anti-tuberculosis drug-induced oxidative stress in kidneys: Role of brahmi as an antioxidant supplement
Sabina Evan Prince, Sherry Joseph Martin, B Udhaya Lavinya, Kavitha Selvanathan, A Geetha
April 2019, 15(62):12-16
DOI:10.4103/pm.pm_481_18  
Purpose: Nephrotoxicity is a known, but rare complication of anti-tuberculosis (TB) therapy and is known to be due to induction of oxidative stress and immune reaction to the drugs. In the current study, we have made an attempt to evaluate the effect of Brahmi in attenuating the oxidative stress caused by isoniazid (INH) and rifampicin (RIF) in the kidneys of Wistar rats. Materials and Methods: The animals were administered INH and RIF with concomitant supplementation of Brahmi for 28 days. The levels of urea, uric acid, creatinine, and acid phosphatase were measured to evaluate kidney function along with catalase, superoxide dismutase, glutathione peroxidase, glutathione-S-transferase, and lipid peroxidation to assess the kidney antioxidant status. In addition, the levels of cholesterol, triglycerides, low-density lipoprotein, high-density lipoprotein, and the cytokine tumor necrosis factor (TNF-α) were also measured. The protective efficacy of Brahmi was compared to that of Silymarin. Results: Brahmi caused a statistically significant (P < 0.05) restoration of kidney function along with an increase in the antioxidant status. This was accompanied by a reduction of TNF-α level to near normal. A histopathological study of the kidney tissues confirmed the protective effect of Brahmi against INH- and RIF-induced oxidative stress in the kidneys. Conclusion: It is evident that Brahmi plays a protective role against anti-TB drug-induced oxidative stress in the kidneys.
  608 0 -
Ultra high performance liquid chromatography-electrospray ionization-mass spectroscopy quantification, xanthine oxidase inhibitory, and antioxidant activity profile of some medicinal plants from Albaha Region
Nasser A Awadh Ali, Iman Mansi, Nafees Ahmed, Saleh Alghamdi, Rajab Abu Alhalawah, Abdulwali Al-Khulaidi, Sirajudheen Anwar
April 2019, 15(62):162-167
DOI:10.4103/pm.pm_514_18  
Background: There is an urgent need to find new xanthine oxidase (XO) inhibitors with few adverse effects and potent activity, not only for treating gout but also to fight diseases associated with XO activity such as cardiovascular diseases, cancer, diabetes, and obesity. Objective: Screening of Saudi medicinal plants for XO inhibitory activity, to quantify the polyphenol–flavonoid content and to study ultra high performance liquid chromatography-electrospray ionization-mass spectroscopy (UHPLC-ESI-MS) profile of compound with best promising XO inhibitory activity among screened extracts. Materials and Methods: Sixteen methanol extracts used traditionally for treating gout and/or rheumatism were screened for total polyphenol and flavonoid contents, XO inhibitory, and antiradical activity via 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay. The degree of XO inhibition was determined by measuring the increase in absorbance at 295 nm associated with uric acid formation. The dose-dependent inhibition profiles of the most active plants were further evaluated by estimating the IC50 values of their corresponding extracts. The most promising XO inhibitory extract, i.e., Dodonaea viscosa extract was subjected to secondary metabolites profiling using UHPLC-ESI-MS in negative ionization mode and LC-MS analysis. Results: Among screened plants, D. viscosa leaves, Punica granatum flowers, Ruta chalepensis leaves, and Solanum incanum fruits exhibited the highest activity with an inhibition of 94.4%, 83.4%, 76.2%, and 65.7%, respectively. Extracts of R. nervosus leaves, P. granatum flowers, and D. viscosa leaves showed the highest antiradical activity in the DPPH assay with IC50 values of 25.8, 27.4, and 71.2 μg/ml, respectively. The LC/MS spectrum of D. viscosa revealed the presence of 13 known compounds along with unknown compounds which belong to flavonoid, terpene, and fatty acid derivatives class. Conclusion: The findings obtained from this study revealed that the methanolic extract of D. viscosa leaf showed the highest xanthine oxidase inhibition activity and therefore is promising species for isolating active compound between the polyphenol and flavonoid content and the XO inhibitory and radical activity.
  592 0 -
Wound healing and antibacterial activities of 2-Pentadecanone in streptozotocin-induced Type 2 diabetic rats
Stepfanie N Siyumbwa, Sophia Ogechi Ekeuku, Farahnaz Amini, Naw May Emerald, Deepali Sharma, Patrick Nwabueze Okechukwu
April 2019, 15(62):71-77
DOI:10.4103/pm.pm_444_18  
Background: Skin regeneration is critical in diabetic wound healing. Anti-inflammatory activity of 2-Pentadecanone (2-P), identified from Marantodes pumilum was reported in our previous studies. Objective: This study was aimed at evaluating 2-P wound healing and anti-bacterial activities. Materials and Methods: Fifteen streptozotocin-induced type 2 diabetic Sprague Dawley rats were divided into negative (N), positive (P), and 2-P treatment (2-P) groups. 2-P aqueous cream was applied daily to 5 mm excision wounds up to day 15. Wound closure rate was assessed by tracing the wound onto plastic sheets and were accessed using a stereoscope microscope. Antibacterial activity (minimal inhibition concentration and disc diffusion) was determined using the modified resazurin assay and Kirby-Bauer method. The skin samples were excised on days 2, 7, and 15 and were analyzed by Hematoxylin/Eosin staining. Semi-quantification analysis was done for re-epithelialization, polymorphonuclear leukocytes (PMNL), and fibroblasts proliferation. Hydroxyproline (collagen deposition) and glutathione (antioxidant) activities were evaluated. Results: The results showed that 2-P possess antibacterial activity against Staphylococcus aureus. It demonstrated significantly higher wound closure compared with negative and positive groups. It showed a higher effect on epithelial cell migration (P < 0.05) and collagen deposition (P < 0.05) compared to negative control group. However, 2-P effect on fibroblast proliferation (P > 0.05) and reduction in PMNL cells (P > 0.05) was not significantly different compared with negative control. Conclusion: It can be concluded that 2-P reveals antibacterial activity, wound closure, collagen deposition, and fibroblast proliferation. Thus, we suggest that 2-P has the potency to be used as an active ingredient in the formulation of a diabetic wound healing cream.
  555 0 -
Cloning, identification, and in silico analysis of terpene synthases involved in the competing pathways of artemisinin biosynthesis pathway in Artemisia annua L
Umara Nissar Rafiqi, Irum Gul, Monica Saifi, Nazima Nasrullah, Javed Ahmad, Prasanta Dash, Malik Zainul Abdin
April 2019, 15(62):38-46
DOI:10.4103/pm.pm_244_18  
Background: Endoperoxide sesquiterpene lactone, artemisinin, is a widely used antimalarial drug. Artemisia annua L. synthesizes this terpenoid and is the only source of artemisinin. In plants, the content of artemisinin is low (0.1–0.8% by dry weight). One of the best approaches to increase artemisinin production is metabolic engineering. Methods: Both the genes were amplified and cloned in Topo vector. Using computational approach, full gene sequencing and a detailed in silico analysis was performed to check the functional and structural properties of these enzymes. Expression patterns of both the genes were assessed at different developmental stages (vegetative, preflowering, flowering, and postflowering stage) of the plant reverse transcription polymerase chain reaction. Results: Deduced amino acid sequence of these genes possessed two important and highly conserved aspartate-rich motifs, and lacks an N-terminal signal peptide, a characteristic of sesquiterpene synthases. Physiochemical properties demonstrated are thermostable. Low hydropathy values ascertain them to be hydrophobic and are active at neutral pH. Structural analysis disclosed that both the proteins possess more α-helices followed by random coils. Ramachandran analysis showed a C-score of −0.35, TM-score of 0.67 ± 0.13 for β caryophyllene synthase model while as C-score of −0.21, TM-score of 0.69 ± 0.12 for β-Farnesene synthase model. Both the proteins contain enormous nitrosylation sites suggesting their functional link through nitrosylation. Gene expression pattern of both the enzymes were upregulated during preflowering and flowering stage. Conclusion: A thorough analysis of these two putative genes in A. annua L paves way to essential insights concerning terpene biosynthesis in general and regulation in artemisinin production in particular. This study also strongly indicates that these two enzymes are developmentally controlled and may have the regulatory effects on the terpene biosynthesis.
  531 0 -
Nutritional value and nephrotoxicity effect of dillenia pentagyna fruit on alloxan-induced diabetic rats
Ranjeet Kumar Yadav, Arpit Azad, SK Srivastava, B Sharma, Mahesh Pal
April 2019, 15(62):118-122
DOI:10.4103/pm.pm_494_18  
Background: Dillenia pentagyna Roxb. (Dilleniaceae) is an endangered medicinal plant, commonly known as “agai” and distributed randomly in the Terai belt of North East states of India. D. Pentagyna extract has been studied for therapeutic possessions. Objective: The proximate parameters and high-performance thin layer chromatography (HPTLC) analysis in the same plant of D. pentagyna of four-time harvest stage during the maturation of fruits were studied and also the antihypoglycemic activity of hydroalcoholic extract of D. pentagyna fruit. Materials and Methods: The proximate compositions were determined by measuring the amount of water removed from the food AOAC method and the chemical fingerprinting was carried out by HPTLC method. An acute oral toxicity study was carried out in healthy male Wister rats (110 g). The dose was finally made to 200 and 400 mg/kg body weight for oral administration after the LD50 estimation. Results: The most phenolic concentration was continuously decrease with during the maturation of fruits. After 21 days of treatment, level of blood glucose, as well as albumin, protein, creatinine, urea, and uric acid were significantly decreased when compared with the diabetic control. Hydroalcoholic (80% alcohol to 20% DW) extracts will be subjected to further extensive studies to isolate and identify their active constituents which are useful for against antidiabetic. Conclusion: D. pentagyna fruit could act as a source of functional compounds for the control of diabetes mellitus.
  529 0 -
Improvement of sexual behavior and ejaculatory parameters by the methanolic extract of Guibourtia tessmannii (Caesalpiniaceae) in high-fat-diet-fed sexually sluggish male rats
Patrick Brice Defo Deeh, Pierre Watcho, Kamalesh K Gulia
April 2019, 15(62):54-61
DOI:10.4103/pm.pm_449_18  
Background: Guibourtia tessmannii (GT) (Caesalpiniaceae) is claimed as a plant having aphrodisiac property as per the African traditional medicine. Objective: This study evaluated the effects of the methanolic extract of GT on sexual behavior and fictive ejaculation in high-fat diet (HFD)-induced sexually sluggish male rats. Materials and Methods: Male Wistar rats fed either on HFD or standard diet (SD) for 16 weeks were monitored for their growth rate and Lee index. At the end of this period, three consecutive copulatory tests were conducted and HFD-induced sexually sluggish rats were selected. Besides Group 1 as time control (SD), drugs or vehicle (veh) were administered orally everyday into three groups of rats as Group 2: HFD (veh), Group 3: HFD + GT (220 mg/kg), and Group 4: HFD + sildenafil citrate (5 mg/kg). Their copulatory activities were tested on day 1, 7, 14, and 21 and the electromyography of bulbospongiosus muscles was assessed for fictive ejaculation on day 22. Results: Treatment with the methanolic extract of GT facilitated sexual behavior by decreasing (P < 0.001) ejaculatory latency within 14 days of treatment that sustained till day 21 compared to the control (HFD) and improving sex drive scores (P < 0.01). The fictive ejaculation parameters were more pronounced in HFD + GT group compared with the HFD group. For instance, after urethral stimulation, the contraction of the bulbospongiosus muscles was significantly increased in HFD + GT group (12.26 ± 7.25) compared to the control group (6.75 ± 0.25). Conclusion: These findings provide robust evidence for the GT treatment in the management of sexual deficits pertaining to obesity.
  525 2 -
Erigeron linifolius attenuates lipopolysachharide-induced depressive-like behavior in mice by impeding neuroinflammation, oxido-nitrosative stress, and upregulation of tropomyosin receptor kinase B-derived neurotrophic factor and monoaminergic pathway in the hippocampus
Chandana Choudhury Barua, Kunjbihari Sulakhiya, Prakash Haloi, Lipika Buragohain, Beenita Saikia, Iswar Chandra Barua, Debesh Chandra Pathak, Shantanu Tamuli, R Elancheran, Xinquo Ren
April 2019, 15(62):92-103
DOI:10.4103/pm.pm_258_18  
Background: Immuno-inflammatory, oxido-nitrosative stress, brain-derived neurotrophic factor-tropomyosin receptor kinase (BDNF-TrkB), and monoaminergic pathways involve in the pathophysiology of depression. Erigeron linifolius (EL) possesses antioxidant, anticonvulsant, antitumor, and hypoglycemic activities. Objective: To investigate the effect of EL hydroalcoholic extract (ELHA) against lipopolysachharide (LPS)-induced depressive-like behavior and neurochemical alterations in mice. Materials and Methods: Mice were pretreated with vehicle, imipramine (10 mg/kg, intraperitoneally [i. p.]), and ELHA (100 and 200 mg/kg, per oral) for 14 days, and depressive-like behavior was induced by LPS (0.83 mg/kg, i. p.). Depressive-like behavior in mice was evaluated by open-field test, forced swimming test (FST), and tail suspension test (TST). Cytokines (tumor necrosis factor-alpha [TNF-α], interleukin-1β [IL-1β], IL-6, and IL-10), BDNF, monoamines (noradrenaline [NA], dopamine, 5-hydroxy), and oxido-nitrosative stress parameters (lipid peroxidation [LPO], glutathione [GSH], GSH peroxidase [GPx], catalase [CAT], superoxide dismutase [SOD], nitric oxide) were measured in the hippocampus (HC). Hippocampal caspase-3, nuclear factor-kappa B phosphor 65 (NF-κB p65), nuclear-related factor 2 (Nrf2), and TrkB and BDNF levels were measured by reverse transcriptase-polymerase chain reaction (RT-PCR) and immunoblotting. Results: Liquid chromatography-electrospray ionization-mass spectroscopy (MS)/MS analysis revealed the presence of ambrosin, friedelin, flavone, scopoletin, and luteolin in ELHA. ELHA showed significant antidepressant-like effect by decreasing the immobility time in FST and TST in LPS-challenged mice. Moreover, ELHA significantly attenuated TNF-α, IL-1β, IL-6, and LPO levels in LPS-challenged mice, whereas LPS-induced decrease in hippocampal IL-10, BDNF, GSH, GPx, CAT, SOD, and monoamine levels was also restored significantly by ELHA. RT-PCR and immunobloting results showed that ELHA significantly attenuated the upregulation of Caspase-3, NF-κB, p65, and Nrf2 and downregulation of TrkB and BDNF levels in the HC of LPS-challenged mice. Conclusion: The findings demonstrated antidepressant-like action of ELHA which could be due to the inhibition of neuroinflammation, oxido-nitrosative stress, and upregulation of TrkB-BDNF and monoaminergic pathways in the HC.
  498 0 -
Formulation and evaluation of wound healing activity of sophorolipid-sericin gel in wistar rats
Snehal V More, Santosh S Koratkar, Navnath Kadam, Sachin Agawane, Asmita Prabhune
April 2019, 15(62):123-127
DOI:10.4103/pm.pm_392_18  
Sericin is a useful by-product of silk processing and is resistant to oxidation and ultraviolet and can absorb and release moisture easily. Sericin has biological activities such as antibacterial, antioxidant, and tyrosinase inhibition. Sophorolipids are microbial extracellular glycolipids produced by resting cells of Candida bombicola. Sophorolipids show excellent skin compatibility and also have antibacterial property. In this study, we have developed a novel formulation consisting of sericin and sophorolipid with calcium alginate as a binding agent. Since both the ingredients are biocompatible and biodegradable, the formulation was tested for wound healing in Wistar rats. A commercial ointment povidone was used as control. The animal group, treated with sericin and sophorolipid cream, showed fast contraction, rapid closure, and healing when compared with control and commercial ointment. These observations were validated with histopathological studies where more fibroblast proliferation, angiogenesis, and keratinization were observed. This is a green, cost-effective formulation for fast wound healing.
  489 1 -
Bioautography-based identification of antioxidant metabolites of Solanum nigrum L. and exploration its hepatoprotective potential against D-galactosamine-induced hepatic fibrosis in rats
Karishma Chester, Sultan Zahiruddin, Adil Ahmad, Washim Khan, Sarvesh Paliwal, Sayeed Ahmad
April 2019, 15(62):104-110
DOI:10.4103/pm.pm_359_18  
Objective: A traditional herb Solanum nigrum L. is well known for the management of different ailments including hepatic disorders. The objective of our study is to identify antioxidant metabolites and bioactive fraction of S. nigrum and to explore their hepatoprotective potential. Materials and Methods: The aerial parts (leaves and berries) of S. nigrum were extracted with hydroethanol- and polarity-based fractionations were performed. Total phenolic (TP), flavonoid content, and thin-layer chromatography (TLC) fingerprints of different extracts were carried out for their quality control and determination of compounds present in them. TLC-based bioautographic assay was carried out to identify the antioxidant metabolites. The hepatoprotective activity of a steroidal glycoalkaloid-enriched fraction of S. nigrum berries was investigated in D-galactosamine (D-GalN)-induced hepatic fibrosis. Hepatic damage was evaluated by assessing enzymatic activities of oxidative markers in serum and liver homogenate and histological study of the liver. Results: The n-butanol fraction of S. nigrum (berries) was found to have the highest value of TP and flavonoids. The treatment of rats with 250 mg/kg crude extract as well as 16 and 25 mg/kg of n- butanol fraction for 10 days was able to normalize the biochemical markers along with liver antioxidative markers in D-GalN treated hepatotoxic rats. The histopathological studies revealed that n- butanol fraction treatment also restored the markers of fibrosis toward a normal level. Conclusion: The n- butanol fraction from S. nigrum berries showed in vitro and in vivo hepatoprotective activity and can be explored after further investigations as a potent phytopharmaceuticals for the management of liver disorders.
  474 0 -
In vitro evaluation of the effects of lycopene on caspase system and oxidative DNA damage in high-glucose condition
Bunyamin Bazyel, Semiha Dede, Sedat Cetin, Veysel Yuksek, Mehmet Taşpinar
April 2019, 15(62):30-33
DOI:10.4103/pm.pm_488_18  
Aim and Background: The present study was planned to investigate the effects of lycopene, on the caspase-dependent apoptosis in high-dose glucose (HG)-treated PC12 cell line. PC12 cells were cultured in vitro. Materials and Methods: HG was prepared as G (250 mM), and lycopene was prepared as L1 (10 μM), L2 (20 μM), and L3 (40 μM). After 6 h of incubation, the cells were exposed to trypsin, and the samples were obtained with freeze/thaw method. Caspase 3, 8, 9; 8-hydroxy-2-deoxyguanosine (8-OHdG); and M30 were determined (enzyme-linked immunosorbent assay). Results: 8-OHdG increased in L3 (P ≤ 0.001), whereas L1 caused a decrease in HG group (P ≤ 0.001). Caspase-3 decreased significantly in L1, L2, and L3G compared to control (P ≤ 0.001) group. Caspase-8 increased significantly in L1, L1G, L2G, and all L3 glucose groups (P ≤ 0.001). There was no difference for Caspase-9. M30 was not affected by L and HG, which decreased significantly (P ≤ 0.001). Conclusion: As a result, it was determined that, when PC12 cell line was treated with HG, lycopene application had effects on caspase enzymes and DNA damage.
  469 1 -
Bioactive compound of Ocimum sanctum carvacrol supplementation attenuates fluoride toxicity in sodium fluoride intoxicated rats: A study with respect to clinical aspect
Kondeti Ramudu Shanmugam, Mavulapati Siva, Sahukari Ravi, Bhasha Shanmugam, Kesireddy Sathyavelu Reddy
April 2019, 15(62):144-149
DOI:10.4103/pm.pm_436_18  
Background: Ocimum sanctum (OS) Linn. commonly known as Holy Basil or Tulsi is an Ayurvedic herb of India. The culinary, medicinal, and industrial importance of this plant led to explore its chemical and pharmacological properties. Objective: The present study was carried out to know the anti-oxidant activity of carvacrol bioactive compound of OS in sodium fluoride (NaF) rats and free-radical scavenging activities of OS. Materials and Methods: Superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), glutathione reductase (GR), glutathione (GSH), GSH-s-transferase (GST), malonaldehyde (MDA), alanine aminotransferase (AAT), aspartate aminotransferase (AST), alkaline phosphatase (ALKP), calcium (Ca), and phosphorus (P4) levels are estimated in all experimental groups. 2,2-Diphenyl-1-picrylhydrazyl (DPPH), hydrogen peroxide (H2O2), and hydroxyl radical activities are analyzed in the ethanolic extract of OS. Results: SOD, CAT, GPx, GR, GSH, activities, and Ca levels depleted and GST, MDA, AAT, AST, ALKP, and P4 levels elevated in NaF intoxicated rats. Whereas carvacrol supplementation normalized all the antioxidant enzymes and hepatic markers in NaF toxicity rats. DPPH, H2O2, and hydroxyl radical of OS showed potent free-radical scavenging activities. In addition, histopathological studies also prove that carvacrol protected the liver tissue from fluoride toxicity in rats. Conclusion: The present study revealed that carvacrol of OS modulated the antioxidant enzymes and hepatic stress markers in NaF rats. Our research study will be helpful in the development of new active principle and nutraceuticals in the area of drug resistance and therapeutic compounds against disease vectors.
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Quantitative assessment of tactile allodynia and protective effects of flavonoids of Ficus carica Lam. leaves in diabetic neuropathy
Khan Dureshahwar, Mohammed Mubashir, Aman Upaganlwar, Jaiprakash N Sangshetti, Chandrashekhar D Upasani, Hemant D Une
April 2019, 15(62):128-134
DOI:10.4103/pm.pm_553_18  
Background: Flavonoids, a group of polyphenols responsible for protective role against many diseased conditions, provide antioxidant activity which is the reason for their medicinal properties. Tactile allodynia is a behavioral biomarker of neuropathy that is well estimated by von Frey filaments and Randall–Selitto test. Objective: Ficus carica Lam. leaves were studied for the conformation of flavonoids in ethyl acetate fraction of methanolic extract (FCEA) using GC-HRMS for the identification of flavonoids. It was analyzed for antioxidant activity by in vitro free radical scavenging activity, performed using 2,2-diphenyl-1-picrylhydrazyl (DPPH) followed by blood glucose-level estimation, evaluation of neuropathic pain, and kidney and liver function tests in diabetic rats. Materials and Methods: The shade-dried leaves of F. carica Lam. were extracted with methanol and after that fractionated using ethyl acetate (FCEA). The characterization of FCEA was established using GC-HRMS. In vitro free radical scavenging activity was performed using DPPH assay. Diabetes was induced using streptozotocin (40 mg/kg/intraperitoneally), and effects of FCEA were studied on blood glucose level, neuropathy markers, and liver and kidney functions of diabetic rats. Results: GC-HRMS results highlighted the presence of quercetin, kaempferol, and chrysin in FCEA with free radical scavenging activity of 78.35% and IC50 value of 5.508 μM. FCEA reduces glucose levels and also shows protective effects in case of diabetic neuropathy as it increases the threshold of withdrawal latency in tactile allodynia and also decreases the serum glutamic-oxaloacetic transaminase, serum glutamic-pyruvic transaminase, blood urea nitrogen, and creatinine levels. Conclusion: The protective effects of FCEA against diabetic neuropathy, hepatoprotective and nephroprotective effects might be due to strong antioxidant property of important flavonoids present which is confirmed in the study.
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Exploring the potential of Desmodium gangeticum (L.) DC. extract against spatial memory deficit in rats
Nilanjan Changdar, Rajesh Kumar Ganjhu, Samita Rijal, Ayush Kumar, Shanchari Basu Mallik, Madhavan Nampoothiri, Rekha R Shenoy, Kiran B Sonawane, Mallikarjuna C Rao, Jayesh Mudgal
April 2019, 15(62):78-83
DOI:10.4103/pm.pm_501_18  
Background: A few published reports demonstrated the neuroprotective effect of Desmodium gangeticum (L.) DC. in an acute model of dementia. Objective: The purpose of the present study was to evaluate the preclinical efficacy of D. gangeticum against chronic dementia when administered prophylactically. Materials and Methods: Chronic spatial memory deficit was induced in rats by aluminum chloride (AlCl3, 10 mg/kg, i.p.). Treatment with hydroalcoholic whole plant extract of D. gangeticum (DG extract) was initiated 2 week before AlCl3 challenge and continued till the 51st day after the challenge, orally at the dose of 400 mg/kg/day. The spatial memory was assessed by Morris water maze test. Hippocampal and frontal cortex acetyl cholinesterase (AChE) and oxidative stress were assessed in diseased rat brains. Results: Chronic administration of AlCl3 produced spatial memory deficit in rats. Memory impairment was manifested in rats as an increase in escape latency and D-quadrant latency whereas a decrease in total time spent in D-quadrant. These behavioral alterations were reversed significantly by the treatment with DG extract. In addition, DG extract significantly increased the island time, indicating memory improvement. DG extract corrected the declined AChE in frontal cortex and altered frontal cortex/hippocampus catalase activity. Phytochemical investigation of the DG extract revealed large content of saponins among the other phytochemicals such as tannins, alkaloids, and flavonoids. Conclusion: These results indicate the possible prophylactic potential of saponin-rich DG extract against chronic memory deficit in rats.
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Cytoprotective effect of Cactus cladode (Opuntia ficus-indica) against chlorpyrifos induced reactive oxygen species in rat hepatocytes: Involvement of heat shock protein 70 and CYP1A1/2 proteins
Hafiz Antar Makeen, Saida Ncibi, Syam Mohan, Abdullah Farasani, Roger Rahmani, Mohammed Al Bratty, Hassan A Alhazmi
April 2019, 15(62):47-53
DOI:10.4103/pm.pm_484_18  
Background: Organophosphorus insecticides are well known to induce hepatotoxicity. One among this is chlorpyrifos (CPF), which is an insecticide inducing various toxicities including in liver. Objective: This investigation focused on CPF-induced oxidative damage in rat hepatocytes primary culture and the protective effect of Cactus cladode aqueous extract. Materials and Methods: Hepatocytes were treated with CPF (50, 75, and 150 μM) and cactus aqueous extract. On treatment for 48 h, mortality within these cells was determined by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide test, reactive oxygen species (ROS) levels were measured by H2DCFDA test. Furthermore, heat shock protein 70 (HSP70) and CYP1A1/2 levels were determined using western blot analysis. Annexin V and 4',6-diamidino-2-phenylindole analysis was run to determine the level of cell death and cytoprotection exerted by CPF and extracts, respectively. Results: The results showed that CPF increases the levels of H2O2 and HSP70 and induces CYP1A1/2 and mortality within these cells. In the other side of this study, the co-treatment of these cells with CPF and Cactus cladode aqueous extract showed a recovery of these parameters. It also has been found that the Cactus cladode aqueous extract has the potential to do cytoprotective effect by preventing necrosis induced by CPF. Conclusion: Taken together, these findings suggest that the toxicity exerted by CPF in hepatocytes are involved with the generation of ROS and the regulation of well-controlled programmed cell death, which could be well protected by the Cactus cladode extract pretreatment.
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Bioavailable curcumin alleviates lipopolysaccharide-induced neuroinflammation and improves cognition in experimental animals
Anu Sunny, Kannan Ramalingam, S Syam Das, Balu Maliakel, IM Krishnakumar, Sibi Ittiyavirah
April 2019, 15(62):111-117
DOI:10.4103/pm.pm_307_18  
Background: Healthy neurons and neurotransmitter levels are necessary for the survival of an organism. Considering the fact that the global incidence of neurological disorders are increasing at an alarming rate; there is a global move toward the development of cost-effective natural neuroprotective agents. Objective: In the present contribution, we hypothesized that the formulations of curcumin capable of delivering curcuminoids in the brain would provide enhanced cognitive effects. In this regard, we investigated the relative efficacy of unformulated curcumin (UC) in comparison with “curcumin-galactomannan complex (CGM), an enhanced bioavailable formulation of curcumin that has been reported to possess improved blood-brain-barrier permeability and tissue distribution (Trademarked as “CurQfen®”). Materials and Methods: Lipopolysaccharide (LPS)-induced neuro-inflammatory animal model was employed for the study. Wistar rats of 180–200 g body weight (aged 3–4 weeks) were grouped as Group I: Vehicle control, Group II: LPS treated (250 μg/kg b.wt.), Group III: CGM (200 mg/kg b.wt.) + LPS (i.p.250 μg/kg b.wt.), and Group IV: UC (200 mg/kg b.wt.) + LPS (i.p.250 μg/kg b.wt.) and treated for 28 days. Results: Behavioral studies (elevated plus maze, radial arm maze, and Y-maze), neurotransmitter levels, and histopathology revealed a statistically significant (P ≤ 0.001) cognitive improvement and reduced inflammation among CGM treated rats as compared to UC treated groups. Conclusion: CGM possesses significant enhanced cognitive effects than UC (IEAC No: DPS/12/2015).
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SHORT COMMUNICATION
Inducible nitric oxide synthase downregulation and cellular antioxidant enzyme enhancing potential of oleanolic acid from Isodon wightii (Bentham) H. Hara against lipopolysaccharide-induced liver damage in bagg albino strain C mice
Madhusudhanan Gogul Ramnath, Ramaraj Thirugnanasampandan, Sadasivam Mathusalini, Gunasekaran Bhuvaneswari, Selvaraj Vasantharani, Palathurai Subramaniam Mohan
April 2019, 15(62):168-172
DOI:10.4103/pm.pm_533_18  
Background: Isodon wightii (Bentham) H. Hara is an aromatic medicinal herb belongs to Lamiaceae which has been considered as a prolific source of diterpenoids with diverse structural and medicinal values. Objective: The objective of the study is to evaluate in vivo anti-inflammatory activity of oleanolic acid from I. wightii against lipopolysaccharide-induced liver inflammation in Bagg albino strain C mice. Materials and Methods: Fractions were obtained using silica gel 60–120 mesh column chromatography, and structural elucidation was done using spectroscopic studies. Hepatoprotection, inducible nitric oxide synthase (iNOS) gene downregulation and the levels of cellular antioxidant enzymes with their respective gene expression were analyzed using reverse transcriptase-polymerase chain reaction (RT-PCR). Results: White amorphous powder (58 mg) was isolated from petroleum ether extract (15 g) eluted with petroleum ether: ethyl acetate mixture (90.5:9.5), and structure has been elucidated as oleanolic acid. 50 μg of oleanolic acid showed effective hepatoprotection and multiple fold increase in the level of antioxidant enzymes such as superoxide dismutase, catalase, glutathione peroxidase, and glutathione S-transferase. RT-PCR analysis revealed that oleanolic acid had a remarkable effect on iNOS downregulation and antioxidant genes upregulation at 50 μg concentration. Conclusion: The present study proves that oleanolic acid possesses liver protecting activity with a positive effect on increased antioxidant enzymes production. The triterpenoid acid and oleanolic acid could be a suitable natural source for preparing hepatoprotective tonics.
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ORIGINAL ARTICLES
The contribution of ionotropic gabaergic and N-methyl-D-Aspartic acid receptors in the antidepressant-like effects of hispidulin
Abeer Abdelhalim, Imran Khan, Nasiara Karim
April 2019, 15(62):62-70
DOI:10.4103/pm.pm_539_18  
Background: Salvia triloba, commonly known as sage belongs to Lamiaceae family, is conventionally used as a brain-enhancing tonic. The purpose of this study was to evaluate hispidulin, a flavonoid isolated from S. triloba for its antidepressant-like effects and to identify its possible mechanisms of action. Materials and Methods: Mouse models of the forced swimming test (FST) and tail-suspension test (TST) were used to assess the antidepressant-like effects of hispidulin. Results: The results showed that hispidulin at the doses level of 1–10 mg/kg, intraperitoneal (ip) exerted significant antidepressant-like effects in both FST and TST. Pretreatment of animals with bicuculline (4 mg/kg, ip; a competitive γ-aminobutyric acid (GABA) antagonist) and N-methyl-D-aspartic acid (NMDA) (75 mg/kg, ip, glutamate receptor agonist) significantly blocked the reduction in immobility time of mice treated with hispidulin (3 mg/kg, ip) in FST. Furthermore, brain GABA levels were significantly decreased by coadministration of hispidulin with bicuculline, whereas glutamate levels were increased with combined administration of hispidulin and NMDA. Moreover, coadministration of sub-effective doses of hispidulin (0.5 mg/kg, ip) and ketamine (0.3 mg/kg, ip) or MK 801 (0.1 mg/kg, ip) also exerted significant antidepressant-like effects in FST. Conclusion: Taken together, these findings suggest that hispidulin possess significant antidepressant-like effects mediated most likely through GABAergic and glutamatergic mechanisms.
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Phytochemical evaluation and cytotoxic potential of chloroform soluble fraction of methanol extract of Thespesia populnea in human breast cancer cell lines
Anu Gopalakrishnan, Bibu John Kariyil, Reni John, PT A. Usha
April 2019, 15(62):150-154
DOI:10.4103/pm.pm_329_18  
Background: Thespesia populnea (Poovarasu), belonging to family Malvaceae, is a tropical evergreen tree found abundantly in the coastal regions of India. It is traditionally known for its hepatoprotective, antitumor, antioxidant, and wound healing activities. Objective: The present study was undertaken to find out different phytochemical principles of chloroform-soluble fraction of T. populnea (CSFTP) using gas chromatography mass spectrometry-mass spectrometry (GCMS-MS) and Fourier-transform infrared (FTIR) spectroscopy and to analyze in vitro cytotoxic potential of the fraction in MDA-MB-231 and MCF-7 human breast carcinoma cell lines. Materials and Methods: Shade-dried bark of T. populnea was extracted using methanol and fractionated to obtain CSF. The fraction was subjected to GCMS, FTIR, and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay. Results: GCMS-MS and FTIR analysis showed the presence of various steroids and fatty acid analogs. Cell growth inhibition was noted for CSFTP in MDA-MB-231 and MCF-7 cell lines. Conclusion: CSFTP revealed the presence of various active principles which could be attributed for its cytotoxic potential.
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Vasorelaxant and antihypertensive activities of citroflavonoids (Hesperidin/Naringenin Mixture): Potential prophylactic of cardiovascular endothelial dysfunction
Amanda Sánchez-Recillas, Nubia Arely González-Rivero, Verenice Barrera-Canto, Maximiliano Ibarra-Barajas, Samuel Estrada-Soto, Rolffy Ortiz-Andrade
April 2019, 15(62):84-91
DOI:10.4103/pm.pm_489_18  
Background: Flavonoids in the Citrus genus have a positive influence in cardiometabolic parameters, preventing cardiovascular diseases (CVDs). The main flavonoids in sweet orange are hesperidin and naringenin. Objective: The aim of this study is to evaluate the cardiovascular effects of mixture of Hesperidin: Naringenin (mix-H:N). Materials and Methods: The relaxant effect and the mechanism of action of mix-H:N were studied on isolated aorta of Wistar rats. Aortic reactivity was determined through concentration-response curves of norepinephrine (NE) and carbamylcholine or carbachol (CCh) after intragastric administration of mix-H:N (150 mg/kg) for 30 days. The antihypertensive effect of a single dose of mix-H:N was studied on spontaneously hypertensive rats (SHR). Results: Mix-H:N produced concentration-dependent relaxation response in Wistar rat's aorta pre-contracted by NE. Inhibitors of NO production and inhibition of extracellular Ca2+ influx caused a significant blockade on the relaxation response to mix-H:N; besides, mix-H:N elicited a vasorelaxant effect on KCl (80 mM)-induced contraction. In addition, oral administration of 150 mg/kg of mix-H:N of SHR rats evoked a significant decrease in systolic and diastolic blood pressure at 5 h and 7 h after administration. Finally, sub-chronic oral administration of mix H:N for 30 days caused ex vivo vascular reactivity modification on NE-induced contraction and CCh-induced relaxation, improving endothelial function. Conclusion: The mix-H:N has the vasorelaxant and antihypertensive effect that may be attributed to an increase of NO production and a blockade of the Ca2+ channels on VSMCs. Furthermore, mix-H:N improves endothelial function of Wistar rats acting as a potential prophylactic against CVDs.
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