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Jul-Sep 2020
Volume 16 | Issue 71
Page Nos. 449-688

Online since Tuesday, October 20, 2020

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ORIGINAL ARTICLES  

Methanolic extract of Mitragyna speciosa Korth leaf exhibits place preference only at higher doses in mice Highly accessed article p. 449
Kamini Vijeepallam, Vijayapandi Pandy, Dharmani Devi Murugan, Murali Naidu Kuppusamy
DOI:10.4103/pm.pm_62_20  
Background: Mitragyna speciosa Korth possesses a wide range of therapeutic benefits, despite having abuse liability. Objectives: The purpose of this research was to investigate the reinforcing properties of M. speciosa Korth leaf extract obtained via methanol extraction using mouse conditioned place preference (CPP) test. Materials and Methods: In CPP study, following baseline preference test (preconditioning score), the mice were subjected to conditioning trials at varying doses of methanolic extract of M. speciosa (MMS, 50, 75, 100, 250, 500, and 1000 mg/kg, p.o.) or reference drugs methamphetamine (0.5 mg/kg, intraperitoneally [i.p.]) and clozapine (1 mg/kg, p.o.) or vehicle controls (1% w/v sodium carboxy methyl cellulose [10 mL/kg, p.o.] and saline [10 mL/kg, i.p.]) followed by a preference test performed under drug-free state (postconditioning score). In addition, the effect of all tested drugs on the spontaneous locomotor activity was assessed. Results: The CPP study results revealed that MMS per se produced a significant place preference only at higher doses (>500 mg/kg, p.o.). Nevertheless, MMS at lower doses (50–250 mg/kg, p.o.) did not induce CPP in mice. In addition, MMS at all tested doses (50–1000 mg/kg, p.o.) did not affect the spontaneous locomotor activity in mice. Conclusion: MMS per se exhibits reinforcing properties at only an increased dose of >500 mg/kg, and therefore, it is best to administer at lower doses (<250 mg/kg) for the potential therapeutic benefits in preclinical studies.
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Apigenin attenuated ethylene glycol induced urolithiasis in uninephrectomized hypertensive rats: A possible role of bikunin, BMP-2/4, and osteopontin Highly accessed article p. 455

DOI:10.4103/pm.pm_83_20  
Background: Nephrolithiasis is a major prevalent health problem of the urinary system worldwide. Apigenin, a plant flavonoid, reported having nephroprotective and antihypertensive properties. Aim: To determine the protective effect of apigenin in experimental ethylene glycol (EG)-induced urolithiasis in uninephrectomized hypertensive animals. Materials and Methods: EG was administration in uninephrectomized rats to induce urolithiasis. Then, the rats were administered with apigenin (5, 10, and 20 mg/kg, p.o.) for 28 days followed by the evaluation of behavioral, biochemical, and histological parameters. Results: Chronic administration of EG induces alterations in the serum and urine parameters (urine output, pH, urine density and dry weight, urea nitrogen, creatinine, uric acid, calcium, sodium, citrate, albumin, glycosaminoglycans, and lactate dehydrogenase), which was significantly inhibited by apigenin treatment. Apigenin treatment attenuated EG-induced elevated cardiac and renal oxido-nitrosative stress. RT-PCR analysis revealed that apigenin inhibited EG-induced modulations in neutrophil gelatinase-associated lipocalin (NGAL), bikunin, kidney injury molecule-1 (KIM-1), inducible nitric oxide synthase (iNOs), endothelial nitric oxide synthase (eNOs), and osteopontin (OPN) mRNA expressions in the kidney. Western blot analysis showed that elevated levels of renal bone morphogenetic protein (BMP)-2/4 proteins were decreased by apigenin treatment. It also significantly inhibited cardiac and renal histological aberrations. Conclusion: Apigenin treatment attenuated EG-induced urolithiasis via the modulation of elevated oxidative stress and altered expressions of KIM-1, NGAL, OPN, bikunin, eNOs, iNOs, BMP-2, and BMP-4 in uninephrectomized hypertensive rats.
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Neuroprotective effect of the essential oil of Lavandula officinalis against hydrogen peroxide-induced toxicity in mice Highly accessed article p. 464
Sarra Akermi, Jackson Roberto Guedes da Silva Almeida, Ahd Khedher, Hafsia Bouzenna, Sabah Dhibi, Anouar Feriania, Afoua Mufti, Amal Daoud, Abdelfattah Elfeki, Najla Hfaiedh
DOI:10.4103/pm.pm_460_19  
Background: Brain is the main organ that manages all other organs and has significant oxygen requirements, which makes it vulnerable to reactive oxygen species, thus causing different neurological disorders. The aim of this work is the evaluation of Lavandula officinalis essential oil neuroprotective effect against hydrogen peroxide (HP)-induced toxicity in mice. Materials and Methods: Essential oil was extracted by hydrodistillation using a Clevenger-type apparatus. Essential oil analysis is performed using gas chromatography-mass spectrometry (GC-MS). Biological activity evaluation carried out the ferric reducing antioxidant power test, deoxyribonucleic acid (DNA) fragmentation assay, the histopathological study of the brain, and determination of antioxidant enzyme activities. Results: Chemical characterization of essential oil using GC-MS identified 47 compounds, accounting for almost 80% of the total oil and indicates the occurrence of monoterpenes and sesquiterpenes. The identified major compounds are pentanone (16.55%), propanal (15.89%), methyl ethyl ketone (13.51%), naphthalene (10.81%), terpinen-4-ol (6.55%), cyclopentanecarboxylic acid (4.77%), and isoborneol (2.27%). This study allowed us to investigate the effects of HP on brain function in Mus musculus adult mice by assessing DNA degradation, cell morphology, oxidative balance and brain weight variation. Furthermore, we have highlighted the beneficial effects of L. officinalis essential oil, which could significantly counteract all these alterations by its active compounds, which are endowed with potent biological activities. Conclusion: We can conclude that HP-induced damage in histomorphological changes in mice brain, significant atrophy, as well as an important alteration of the genetic expression.
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Comparative study of the pharmacokinetic parameters for salidroside in normal and estrogen-deficient female rats after oral administration of an aqueous extract of Fructus Ligustri Lucidi using a validated ultra-performance liquid chromatography mass spectrometry/mass spectrometry method p. 471

DOI:10.4103/pm.pm_278_19  
Background: Salidroside, one of the main active ingredients in Fructus Ligustri Lucidi (FLL), is well demonstrated to exert anti-osteoporotic effect. However, the plasma pharmacokinetic profile of salidroside in FLL in estrogen-deficient rats remains unknown. Objective: The objective was to develop a sensitive, rapid, and accurate ultra-performance liquid chromatography-mass spectrometry (UPLC-MS) method for the determination of the pharmacokinetics profile of salidroside after oral administration of FLL aqueous extract in normal and ovariectomized (OVX) rats. Materials and Methods: OVX and normal rats were orally administrated with FLL at a bolus of 7 g/kg. Plasma samples were precipitated by methanol, and the supernatant was chromatographed by a Waters BEH C18column with a gradient elution of ammonium acetate and acetonitrile. Quantification was carried out on the electrospray ionization, positive multiple reaction monitoring modes. Results: The lower limit of detection was 50 ng/mL, and the dynamic linear range was 50–30,000 ng/mL with a value of R2 > 0.99. The intra- and inter-day precisions were lower than 14.67%, and accuracy was in the range of 99.29%–103.37%. The recovery of salidroside ranged from 88.90% to 101.78%, with the matrix effect ranging from 85.53% to 100.45%. The t1/2, MRT0–∞, and apparent volume of distribution for salidroside increased in OVX rats. Conclusion: A sensitive, accurate, and rapid method was successfully established and validated for the determination of plasma characteristics of salidroside in Sprague–Dawley (SD) rats. The results suggest that ovariectomy could interfere with salidroside metabolism in SD rats.
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Narirutin suppresses M1-related chemokine interferon-gamma-inducible protein-10 production in monocyte-derived M1 cells via epigenetic regulation p. 479
Chung-Hsiang Li, Lin Wang, Chang-Hung Kuo, Yi-Ting Chen, Mei-Lan Tsai, Chih-Hsing Hung
DOI:10.4103/pm.pm_105_20  
Background: Flavonoids are groups of natural phytonutrients found in fruits and vegetables that have recently become popular because of their anti-oxidation and anti-inflammatory ability. Narirutin is a flavanone which has been proven to have anti-inflammation effects, although its fundamental mechanisms are not understood. Objective: We try to investigate this anti-inflammatory effect of narirutin in human monocytic THP-1-derived M1 macrophage cells. Materials and Methods: To confirm our hypothesis, human THP-1 cells (1 × 106 cells/mL) were initially treated with 20 ng/mL phorbol 12-myristate 13-acetate (PMA) for 24 h. The PMA-differentiated THP-1 cells were treated with various levels of narirutin 2 h before lipopolysaccharide stimulation; after that, the cells were cultured for 24–48 h and then examined. The concentration of interferon-gamma-inducible protein-10 (IP-10) was measured using enzyme-linked immunosorbent assay. Epigenetic regulation mechanisms were explored by chromatin immunoprecipitation assay. Results: Narirutin significantly suppressed IP-10 production in M1 macrophage cells, and the suppressing effect was partly reversed by the estrogen receptor antagonist, the aryl-hydrocarbon receptor antagonist, the peroxisome proliferator-activated receptor (PPAR)-α antagonist, and the PPAR-γ antagonist. We also found that narirutin-induced IP-10 suppression can be modulated by both histone H3 and H4 acetylation. Conclusion: Our study suggests the potential of narirutin for the treatment of inflammatory disease by suppressing IP-10.
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Effect of diosgenin on T-helper 17 cells in mice with collagen-induced arthritis Highly accessed article p. 486
Yaxian Gao, Yongwei Wang, Hongru Song, Yachun Guo, Enhong Xing, Xiaofei Zhao, Wei Li, Junxia Zhang, Chunyan Yu
DOI:10.4103/pm.pm_426_19  
Background: Diosgenin, obtained from Dioscorea nipponica Makino, possesses anti-inflammatory properties. Objective: The objective of this study is to explore the therapeutic effects of diosgenin against collagen-induced arthritis (CIA) in DBA/1J mice. Materials and Methods: The CIA model was established using chicken type II collagen-immunized DBA1/J mice (0.1 mL/mouse), with the same stimulus repeated on day 21. Next, the CIA mouse model was selected, and inguinal lymph nodes were excised to obtain a single-cell suspension. Using CCK-8, control group, model group, and high-/medium-/low-dose diosgenin group were set based on drug toxicity. Next, the ratio of T-helper 17 cells (Th17) to CD3+ T cells was analyzed using flow cytometer. The levels of interleukin 17 (IL)-17 and IL-6 were determined by the enzyme-linked immunosorbent assay. The expression of RORγt was determined using the reverse-transcription quantitative real-time polymerase chain reaction and Western blot analysis. Results: An in vitro inhibitory rate of ≥ 70% was set as the cutoff value to select the high (25 μmol/L), medium (12.5 μmol/L), and low (6.25 μmol/L) doses of diosgenin. The Th17 cell ratio in CD3+ T cells, the levels of IL-17 and IL-6, and mRNA and protein expression of RORγt showed significantly increased tendency in the model group compared with the control group (P < 0.01 and/or P < 0.05). Diosgenin significantly decreased the Th17 cell ratio in CD3+ T cells, IL-17 and IL-6 levels, and RORγt expression when compared with the model group (P < 0.01 and/or P < 0.05). Conclusion: Diosgenin exhibited an antiarthritic effect in CIA mice by downregulating the differentiation of Th17 cells.
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Immunostimulatory activities of a high molecular weight fraction of Cynanchum auriculatum royle ex wight root obtained by ultrafiltration p. 493
Mi Jang, Tae-Gyu Lim, Eunjung Lee, Hee-Do Hong, Young Kyoung Rhee, Yun Ji Lee, Jeong Hoon Lee, Chang-Won Cho
DOI:10.4103/pm.pm_449_19  
Background: The root of Cynanchum auriculatum Royle ex Wight has been traditionally used as a folk medicine in the eastern Asia. Objectives: In this study, we explored the immune-enhancing effect of the high-molecular weight fraction of C. auriculatum (CAHF) root extracts in RAW 264.7 macrophages and cyclophosphamide (CYP)-induced immune-suppressed mice. Materials and Methods: To obtain the CAHF, ultrafiltration was performed using 30-kDa molecular weight cutoff polyethersulfone membranes in a cross-flow microfiltration system. In vitro and in vivo experiments were performed to evaluate the immunostimulatory effects of CAHF. Results: CAHF increased the production of nitric oxide and immunostimulatory cytokines (interleukin-6 and tumor necrosis factor-α) in recombinant interferon-γ primed macrophages. In addition, the phagocytic activity of macrophage cells was enhanced by CAHF. Furthermore, CAHF treatment markedly restored the decreased body weight, immune organs (spleen and thymus) weight, natural killer cell activity, and T- and B-lymphocyte proliferation in CYP-induced immune-suppressed mice. Conclusion: These results suggest that CAHF is an effective immunostimulatory agent.
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Anticancer activity of Juniperus procera grown in southwestern region of Saudi Arabia on human oral squamous cell carcinoma cell lines p. 499

DOI:10.4103/pm.pm_28_20  
Background: One of the popular ancient plants used to treat various diseases such as hyperglycemia, hepatitis, jaundice, bronchitis, and pneumonia has been Juniperus procera (JP). JP is abundantly seen in the region of Al-Baha, Saudi Arabia, and is being used as a medicinal plant traditionally by local healers. Objectives: The objective was to evaluate the anticancer properties of JP from Al-Baha region on SCC-9 cells. Materials and Methods: Colorimetric assays such as 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay, sulforhodamine B assay, and lactate dehydrogenase (LDH) assay were performed to check the cytotoxicity induced and cell viability. Assays evaluating cellular events such as apoptosis and cell cycle were also performed. Results: MTT assay revealed IC50value of 201.6 μg/ml. The cancer cells primed with increasing concentrations of JP displayed an enhanced emission of LDH at elevated concentrations contrasted to cells which were not treated. The samples of JP pooled fraction (PF) (6–9) treated at 160 μg/ml and 320 μg/ml concentration showed 8.39% and 23.37% and 17.35% and 20.89% in early and late phases of apoptosis, respectively. Sample P (PF 6-9) at 160 μg/ml and 320 μg/ml has induced a G2M phase arrest of up to 21.06% and 26.94%. Deoxyribose nucleic acid damage was compared in tested concentrations of sample JP with untreated control cells in SCC9 cells. SCC9 cells that were treated with sample JP PF 6–9 showed the olive moments 23.22 and 37.30 at concentrations 160 μg/ml and 320 μg/ml, respectively. Studies of gene expression showed that increased concentrations of JP triggered the development of caspases and p53. Conclusion: The bioactive compounds found in JP were effective and potent against the SCC-9 cancer cells.
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Protective effects of Gracilaria lemaneiformis extract against ultraviolet B-induced damage in HaCaT cells p. 510

DOI:10.4103/pm.pm_437_19  
Background: Gracilaria lemaneiformis is an edible red marine macroalga that contains various active components including phycoerythrin, polysaccharides, and phenolics. In our previous work, crude ethanolic extracts of G. lemaneiformis exhibited potential antioxidative and anti-photoaging activities. Therefore, in this study, we aimed to further investigate the antioxidative and anti-photoaging activities of different fractions of G. lemaneiformis (n-hexane, ethyl acetate, and aqueous fractions [HF, EAF, and AQF, respectively] using ultraviolet B (UVB)-induced HaCaT cells. Materials and Methods: Solvents with different polarity were used to fractionate crude ethanolic extract of G. lemaneiformis. The physicochemical properties of HF, EAF, and AQF were detected by gas chromatography–mass spectrometry, Fourier transform-infrared spectroscopy, and high-performance liquid chromatography–mass spectroscopy. The antioxidant and antiapoptotic effects of the fractions were evaluated by estimating the levels of reactive oxygen species, antioxidant enzymes, and mitochondrial membrane potential (MMP) in UVB-exposed HaCaT cells. Results: According to our results, fatty acids, chlorophyll-a, and soluble polysaccharides were, respectively, present in the HF, EAF, and AQF. Furthermore, both EAF and AQF decreased the UVB-induced apoptosis by decreasing MMP. These results also suggest that EAF and AQF provide cytoprotective effects by activating the antioxidant enzymes. The soluble polysaccharides of AQF and chlorophyll-a of EAF were positively correlated with antioxidant activity. In addition, AQF exhibited stronger antioxidant and anti-photodamage properties than that of other fractions in UVB-radiated HaCaT cells. Conclusion: The results of this study indicated that water-soluble polysaccharides from G. lemaneiformis may be suitable to use as a natural anti-photodamage agent.
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Involvement of toll-like receptor 2/myeloid differentiation factor 88/nuclear factor kappa B/NLR family pyrin domain-containing 3 signaling pathways in the hepatoprotective effect of Lagotis brachystachys in rats with alcoholic liver disease p. 518

DOI:10.4103/pm.pm_557_19  
Background: Excessive consumption of alcohol is ranked as one of the leading causes of death from alcoholic liver disease (ALD). Our previous study demonstrated that the whole extracts of Lagotis brachystachys decreased acute hepatic injury in rats. However, the compounds of the extracts that are responsible for the hepatoprotective activity and their underlying mechanism are still unknown. Objectives: The objective of the study was to evaluate the hepatoprotective effect of whole extracts of L. brachystachys against chronic alcohol-induced ALD in rats. Materials and Methods: Different polar compounds of ethanolic extract from L. brachystachys were orally administered to rats that underwent 8 weeks of alcohol exposure. Results: The histological evaluation of rat liver showed that the rats exposed to 30% and 50% ethanolic extracts of L. brachystachys had significantly less formation of lipid droplets and showed less inflammatory infiltration than that of control rats with ALD. The extracts also inhibited alcohol-induced elevation of serum lipid peroxidation levels. In addition, L. brachystachys restored the levels of antioxidants and inhibited the alcohol-induced activation of the hepatic Toll-like receptor 2 (TLR2)/myeloid differentiation factor 88 (MyD88)/nuclear factor kappa B (NF-κB)/NLR family pyrin domain-containing 3 (NLRP3) signaling pathway, thereby decreasing the release of interleukin (IL)-1β. Conclusion: Our data revealed that L. brachystachys showed hepatoprotective effect against chronic alcohol-induced hepatic injury by decreasing the levels of lipid peroxidation and oxidative stress and by inhibiting the inflammatory processes. The extracts decreased the release of IL-1β via inactivation of the hepatic TLR2/MyD88/NF-κB/NLRP3 signaling pathway.
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Honey as a solvent for the green extraction, analysis, and bioconversion of daidzin from Pueraria candollei var. mirifica root p. 524

DOI:10.4103/pm.pm_74_20  
Background: Honey has been widely used as a traditional vehicle of herbal medicines. Honey behaves as a natural deep eutectic solvent (NADES) containing β-glucosidase; therefore, it can be used for the extraction and bio-activation of the bioactive compounds of herbs. Objectives: This study aims to apply honey (H-NADES) and a sugar-based NADES (S-NADES) for the extraction, analysis, and bioconversion of daidzin from Pueraria candollei var. mirifica (PM) root. Materials and Methods: Various concentrations of H-NADES and S-NADES (water:sucrose:glucose: fructose, 18:3:18:22 by weight) were used as solvents for extraction. Indirect competitive enzyme-linked immunosorbent assay (icELISA) was developed and validated for monitoring the extraction efficacy. The catalytic reactivity against daidzin of β-glucosidase purified from honey was investigated. Results: Using NADESs as solvents, icELISA was suitable for the reliable determination of daidzin with high sensitivity (1.95–125 ng/mL), specificity (% cross-reactivity ≤ 2.60), and accuracy (98.3-106% daidzin recovery). Daidzin at a concentration of 75.8 ± 3.67 μg/mL was extracted using 50% (v/v) S-NADES, which was the most effective for the extraction compared to H-NADES, water and ethanol. In addition, daidzin was converted to daidzein by honey β-glucosidase. Conclusion: Both S-NADES and H-NADES were useful for the extraction, analysis, and bioconversion of daidzin, and β-glucosidase from honey might enhance the oestrogenic activity and bioavailability of PM phytochemicals.
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Chlorogenic acid in Viscum album callus is a potential anticancer agent against C6 glioma cells p. 531

DOI:10.4103/pm.pm_479_19  
Background: Chlorogenic acid (CA), a polyphenolic component of fruits, vegetables, coffee, wine, and olive oil, has beneficial effects on human heath, including antioxidant and anticancer effects. However, its precise effects on glioma have not been examined. Objective: Our study aimed to explore the anticancer effects of CA obtained from Viscum album callus on C6 glioma cell migration and proliferation. Materials and Methods: Anticancer potency was analyzed by the 2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide salt assay to assess the ability to inhibit cell growth and proliferation. Cell mobility was investigated based on the Boyden chamber and the scratch wound healing assay. Factors involved in cell cycle progression were evaluated by mRNA and protein expression. Cell death was determined by staining with specific dyes and fluorescence microscopy. Results: CA significantly reduced C6 glioma cell proliferation and migration. Furthermore, it induced reactive oxygen species generation and apoptotic cell death. Treatment with CA also suppressed extracellular signal-regulated kinase ½ (ERK½) phosphorylation and the gene expression of cyclins E and A. Conclusion: Our results show that CA may regulate glioma cell migration and proliferation via modulation of ERK½ phosphorylation and cell cycle regulation. Thus, it might be a potent anticancer agent in preventing progression of glioma.
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Identification of Polygonatum odoratum based on support vector machine p. 538

DOI:10.4103/pm.pm_410_19  
Background: The dried rhizome of Polygonatum odoratum (Mill.) Druce has been widely used in traditional medicinal preparations in China, Japan, and Korea. In China, it is distributed in Hunan, Guangdong, and Liaoning provinces, and its quality differs from habitat to habitat. In addition, P. odoratumhas some adulterants, such as Polygonatum inflatumKom, Polygonatum prattii Baker, and Polygonatum cyrtonema Hua. The morphological traits and chemical composition of the aforementioned adulterants have many similarities with those of P. odoratum. Therefore, it is possible that people often use adulterants instead of P. odoratum for clinical treatment. Objectives: We aimed to establish a reliable and accurate classification model of P. odoratum based on the support vector machine (SVM) and identify it from different habitats; we also aimed to identify its adulterants. Materials and Methods: In this study, we first determined the ultraviolet (UV) absorption spectrum of the water extract of the rhizome from 162 samples (including P. odoratum from Hunan, Guangdong, Heilongjiang, Yunnan, and Liaoning Provinces and adulterant species including P. inflatum, P. prattii,P. cyrtonema, and Disporopsis pernyi (Hua) Diels) by UV-visible spectrophotometry. The UV absorption data were preprocessed with the SVM model before establishing the habitat and other details. Results: According to our results, the SVM model showed a prediction accuracy of 100%. The model accurately identified five different habitats and four different adulterants of P. odoratum. Pretreatment of samples with UV spectrum might be useful in the accurate identification of P. odoratum. Conclusion: The SVM model seems very prospective in identifying herbs with multiple habitats and its adulterants.
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Preparation, characteristics, and antioxidant activity of the selenium nanoparticles stabilized by polysaccharides isolated from Grateloupia filicina p. 543

DOI:10.4103/pm.pm_405_19  
Background: Selenium is an essential trace element which is of fundamental importance to human health. Compared to organic and inorganic selenium, selenium nanoparticles (SeNPs) display unique biological and physicochemical properties. Objectives: In this study, we aimed to optimize the extraction parameters of polysaccharides of Grateloupia filicina (GFPs) and investigate the antioxidant activity of SeNPs stabilized by GFPs. Materials and Methods: GFPs were extracted using hot water. The extraction parameters were optimized by performing an orthogonal experiment. SeNPs were prepared under mild conditions using GFP as the modifier and the stabilizer. A scanning electron microscope (SEM) was used to characterize the prepared GFPs-SeNPs. The antioxidant activities of GFPs, SeNPs, and GFPs-SeNPs were compared by measuring 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity, hydroxyl radical scavenging activity, superoxide anion radical scavenging activity, and via reducing power assay. Results: The optimal conditions for the extraction were determined as follows: extraction time 5 h, extraction temperature 90°C, and the ratio of water to raw material 100 mL/g. Under these conditions, the yield of GFPs was 50.34% ±0.27%. GFP-decorated SeNPs with an average diameter of 100 nm were synthesized. GFPs-SeNPs showed a higher antioxidant activity compared to GFPs and SeNPs alone. Conclusion: Hot water extraction was found to be efficient in the process of extraction of polysaccharides of G. filicina. GFPs can significantly enhance the antioxidant activity of SeNPs as modifier and stabilizer. GFPs-SeNPs is an efficient radical scavenger and may be explored as a novel antioxidant agent for use in the fields of foodstuff and medicine.
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Anticancer effects of Calotropis procera latex extract in mcf-7 breast cancer cells p. 550

DOI:10.4103/pm.pm_156_20  
Background: Calotropis procera is a wild growing medicinal plant with many pharmacological properties, arising mainly from its latex, which contains many biologically active compounds, including cardiac glycosides. Objectives: The present study was conducted to isolate a cardiac glycosidal (CG) extract from the latex of C. procera and to assess its potential in inducing anticancer effects on breast cancer cells (MCF-7). Materials and Methods: Cytotoxicity was evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay and morphological changes observations. The generation of intracellular reactive oxygen species (ROS) was evaluated both qualitatively and quantitatively. Flow cytometry technique was used to evaluate apoptosis and autophagy was determined by fluorescence microscopy and Western blotting. Results: The extract significantly (P < 0.05) inhibited the proliferation of MCF-7 cells and that this effect increased in line with concentration. Systemic changes in the morphology of treated cells when compared with control cells were observed. ROS levels were increased by about 1.5 and 1.95-fold at the highest concentration of 75 μg/ml after 12 and 24 h of treatment, respectively. A significant (P < 0.05) increase in the percentage of early and late apoptotic cells were recorded. Autophagy induction in treated MCF-7 was confirmed with the presence of acidic vesicular organelles. Finally, the change in the intracellular localization of light chain 3 (LC3) protein was determined by Western blotting using primary antibodies. A maximum of a 1.76-fold increase was observed in the expression level of the LC3 marker protein. Conclusion: These findings suggest that CG extract increased the levels of intracellular ROS resulting in the induction of cytotoxicity, apoptosis, and autophagy in MCF-7 cells.
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Isovitexin, A new metabolite, was found in the metabolites of co-cultured five flavonoids isolated from Ziziphus jujuba Mill var. spinosa seeds by rat intestinal flora p. 557

DOI:10.4103/pm.pm_454_19  
Background: Ziziphus jujubaMill var. spinosa seeds (ZSS) is one of the most popular traditional Chinese herbs. It shows several pharmacological effects, such as anti-anxiety, antidepressant, neuroprotection, and cardiotonic, and prevents insomnia. The primary biologically active flavonoids derived from ZSS are 6´´´-feruloyl spinosin, 6´´´-p-coumaroyl spinosin, spinosin, swertisin, and isovitexin. Objectives: The primary objective of this study was to investigate the mechanism of degradation of co-cultured flavonoids, namely 6´´´-feruloyl spinosin, 6´´´-p-coumaroyl spinosin, spinosin, swertisin, and isovitexin, in the intestinal flora of rats under in vitroconditions. Materials and Methods: In this study, we co-cultured the five flavonoids in the intestinal flora of rats under in vitro conditions and determined the degradation of these five flavonoids via high-performance liquid chromatography with tandem mass spectrometry (MS/MS). Results: The degradation rate of the 6´´´-feruloyl spinosin, 6´´´-p-coumaroyl spinosin, and swertisin was affected by the concentration of the sample and conforms to the first-order kinetic model. According to our results, the 6´´´-feruloyl spinosin and 6´´´-p-coumaroyl spinosin may be degraded to spinosin and swertisin, and spinosin continued to decompose into swertisin. These results were consistent with their individual experiments. Moreover, by comparing the structures of isovitexin and swertisin, isovitexin was first found to be the product of the seventh demethylation reaction of the flavonoid core structure, demonstrating that isovitexin is a metabolite of ZSS flavonoids following spinosin and swertisin. Conclusion: Taken together, the results of this study explain the metabolic and interrelation of these five main flavonoids in vitro.
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Chemical constituents from Saussurea pachyneura p. 564

DOI:10.4103/pm.pm_453_19  
Background: Genus Saussurea is famous for its use in traditional Chinese medicine. Objectives: Saussurea pachyneura, a plant from genus Saussurea, was studied for its secondary metabolites. Materials and Methods: The air-dried whole plant material was extracted by ethanol. The compounds from the ethanolic extract were isolated and purified by silica gel, octadecylsilyl-silica gel, and Sephadex LH-20 column chromatography. Their structures were identified based on the spectral analysis. Results: A total of 13 compounds, including a new long-chain fatty acid ester (4), were isolated and identified from the ethanolic extract of S. pachyneura. Conclusion: This is the first study to report the phytochemical analysis of S. pachyneura and also the first study to isolate the compounds from ethanolic extract of the whole plant material of S. pachyneura.
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Synthesis, characterization, and biological activity of silver nanoparticles synthesized from Aristolochia bracteolata lam. p. 568

DOI:10.4103/pm.pm_313_20  
The aim of this research is to analyze the antimicrobial activity and cytotoxicity of biologically synthesized silver nanoparticles against A549 cancer cell line. Silver nanoparticles were synthesized from the leaf extract of Aristolochia bracteolata. For the characterization of silver nanoparticles, Fourier transform infrared, high-performance liquid chromatography, and ultraviolet techniques were adopted. The silver nanoparticle was found to show better antimicrobial activity when compared to the crude extract of the plant. Noteworthy cytotoxic effects against A549 cell line were shown by the nanoparticles derived from the plant extract. Following the potential of biologically synthesized silver nanoparticles, they can be a valuable addition to the treatment of human laryngeal carcinoma and can be categorized as a potent anticancer agent.
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Isoliquiritigenin induces apoptosis through caspases and reactive oxygen species signaling pathways in human bladder cancer cells p. 574

DOI:10.4103/pm.pm_21_20  
Background: Isoliquiritigenin (ISL) is a flavonoid isolated from the roots of various species of licorice plants. Objectives: Mechanisms underlying ISL-induced cell death were investigated in 5637 human bladder cancer cell line. Materials and Methods: Cell viabilities were measured with 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide and cell counting kit-8 assay. Cell cycle analysis, caspase activity assay, western blotting, and reactive oxygen species (ROS) assay were also used to investigate the anticancer effects of ISL on 5637 cells. Results: ISL (100–500 μg/ml) inhibited cancer cell proliferation and increased sub-G1 cell cycle phase ratios. ISL-induced cell death resulted in reduced Bcl-2 and increased Bax. ISL also activated caspase-3 and -9 and increased the levels of intracellular ROS generated. In addition, TG100-115 (transient receptor potential [TRP] melastatin 7 inhibitor) and tranilast (TRP vanilloid 2 inhibitor) each exerted a synergistic effect with ISL on ISL-induced apoptosis. Conclusion: These findings suggest that ISL causes apoptosis in 5637 cancer cell line. Therefore, ISL may be a potential anticancer drug for treating bladder cancer and a good anticancer supplement.
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Analgesic, anti-inflammatory, and central nervous system depressant activities of Monochoria hastate (L.) Solms. in animal models p. 580

DOI:10.4103/pm.pm_463_19  
Background: Monochoria hastate (L.) Solms. is extensively used as traditional medicine despite the fact that no scientific data has yet been published revealing its biological activity in vivo. Objectives: The objective of the study was to investigate the pharmacological effect of crude leaf extracts of M. hastate in mice model, justifying its medicinal use and evaluating its safety and efficacy as a crude drug. Materials and Methods: Crude extracts were prepared using methanol and ethyl acetate as solvent. Analgesic activity was evaluated by writhing and formalin-induced licking bioassay. Anti-inflammatory activity was examined by detecting its effect on carrageenan-induced edema. Effect on the central nervous system (CNS) was tested by monitoring movements of test animals by open-field and hole-cross assay. Results: Oral administration (400 mg/kg) of methanol and ethyl acetate extract resulted in 93% and 92% reduction of writhing (P < 0.001) compared to the 87% by diclofenac, whereas ethyl acetate extract caused maximum inhibition (91%) of licking response. Both the extracts manifested marked (P < 0.05) reduction in edema diameter, but the effects were less significant than the standard. Open-field and hole-cross test demonstrated significant (P < 0.001) suppression of motor activity in the treated group. The resulting movements 120 min after oral administration of methanol and ethyl acetate extract (400 mg/kg) and diazepam (1 mg/kg) were 205.8 ± 33, 1.4 ± 0.5, 217 ± 28, 1.0 ± 0.3, 191.2 ± 25, and 2.0 ± 0.7, respectively. Conclusion: These findings provide substantial evidence that leaf extracts of M. hastate possess analgesic, anti-inflammatory, and CNS depressant activity in mice explaining some of its use in traditional medicine.
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Protective effect of caffeic acid phenethyl ester against acute and subchronic mice cardiotoxicity induced by cyclophosphamide alone or plus naproxen p. 585

DOI:10.4103/pm.pm_159_20  
Background: The limiting factor in the use of cyclophosphamide (CYP) in cancer chemotherapy is its induced oxidative cardiotoxicity. Objectives: This study aims to investigate the possible protective effect of caffeic acid phenethyl ester (CAPE) in the co-administration of CYP and naproxen (NAP) with acute and subchronic treatments in mice. Materials and Methods: Male BALB/c mice were divided into two phases of acute (24 h) and subchronic (30 days) treatments, of which seven groups each were used. Two groups from both acute and subchronic treatments represented untreated controls and CAPE groups, while others were CYP, NAP, CYP+NAP, CYP+CAPE and CYP+NAP+CAPE groups for both treatments. The activity of the cardiac antioxidative enzyme catalase was measured. The levels of cardiac reduced glutathione (GSH), protein carbonyl and malondialdehyde (MDA) were also assayed. In addition, histopathology of the heart tissues and immunohistochemistry of endothelial nitric oxide synthase (eNOS) expression were evaluated. Results: Our results showed that catalase and GSH were significantly decreased in all subchronic treatments. Furthermore, protein carbonyl and MDA were increased in both acute and subchronic treatments. Histopathological examination showed hypertrophic cells induced by CYP, NAP, and in combination. Moreover, CYP, NAP and in combination, significantly reduced eNOS levels. However, CAPE significantly prevented changes induced by CYP and NAP in both treatment groups. Conclusion: These observations highlight the protective potentials of CAPE in CYP-NAP-induced cardiotoxicity.
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Profiling of antiviral and antioxidant phytochemicals of Pterocephalus frutescens hochist. using high-resolution ultra-performance liquid chromatography/quadrupole time-of-flight mass spectrometer p. 592

DOI:10.4103/pm.pm_558_19  
Background: Medicinal plant extracts and herbal preparations are complex mixtures of active and ballast substances, which may contain in frequently up to several hundreds of different constituents, with no exactly defined structures at which chromatography undoubtedly is a fundamental tool to overcome the challenges of these phytoanalytics. Aim of the Study: To investigate Pterocephalus frutescens aerial part extracts biologically and chemically and the correlation of biologically active constituents with their corresponding activities. Materials and Methods: Ultra-performance liquid chromatography/quadrupole time-of-flight mass spectrometer with principal component statistical analysis was used for the identification of phytoconstituents, while spectrophotometer was used for quantitative analysis and both qualitative and quantitative antioxidant effects using stable radical diphenylpicrylhydrazyl, and the cytopathic effect inhibition assay was used for antiviral screening. Results and Conclusion: P. frutescens extracts exhibited antioxidant and antiviral activities along with tentative identifying of 46 compounds from which 38 compounds were correlated with either antiviral and/or antioxidant activities, besides the quantification of phenolic, flavonoid, phenylethanoid, and iridoid contents.
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Polygonum chinense water decoction lessens acute lung injury in mice induced by influenza virus p. 600

DOI:10.4103/pm.pm_206_19  
Aim/Background: In traditional Chinese medicine, Polygonum chinense has been used to treat influenza. However, little information is available from current studies regarding the anti-influenza pharmacological activities of P. chinense water decoction (PCWD) and its underlying mechanisms. The present study aimed at investigating the treatment of PCWD on acute lung injury (ALI) induced by the H1N1 influenza A virus (IAV) and its underlying mechanisms. Materials and Methods: Mice were infected with IAV. PCWD (300 and 600 mg/kg/day) and ribavirin (100 mg/kg/day) were orally administrated to mice. Mice survival rate was observed for 15 days after inoculation. On day 5 after virus infection, serum and lung tissues of mice were collected for the analysis of lung index, virus titers, cytokines, histopathology, and immunohistochemistry. Results: PCWD significantly lessened ALI and improved survival rate induced by H1N1. PCWD also decreased the 5th day lung index and ameliorated the injuries, inflammatory cells infiltration, and lung edema. PCWD reduced the level of tumor necrosis factor-α and interleukin-6. PCWD obviously decreased hemagglutinin titer in the lungs. Immunohistochemistry showed that PCWD obviously inhibited TLR-4 and p-NFκB p65 expression. Conclusion: PCWD can alleviate ALI induced by IAV through inhibiting inflammation.
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Fisetin attenuates gastric mucosal lesions through modulating nuclear factor-kappa B and peroxisome proliferator-activated receptor-γ in rats p. 605

DOI:10.4103/pm.pm_4_20  
Background: Nowadays, the use of plant extracts is increasing in the world for the prevention and treatment of ulcer. Objective: The objective of this study was to explore the underlying mechanism of action of fisetin on ethanol-induced gastric ulcer model. Materials and Methods: In this study, gastric mucosal lesions were induced by ethanol in rats. Five groups of rats were formed based on the treatment administered: model group (model), omeprazole (40 mg/kg) group (omeprazole), high-dose fisetin group (100 mg/kg, H-fisetin), medium-dose fisetin group (50 mg/kg, M-fisetin), and low-dose fisetin group (25 mg/kg, L-fisetin). Interleukin (IL)-1β, IL-6, and tumor necrosis factor (TNF)-α levels were assessed in serum. The expression of peroxisome proliferator-activated receptor (PPAR)-γ, nuclear factor-kappa B (NF-κB), and p38-mitogen-activated protein kinase (p38-MAPK) in the gastric mucosa was also measured. Results: In the case of the high-dose fisetin group, the level of TNF-α, IL-1β, and IL-6 decreased from 9.57 pg/mL to 5.19 pg/mL, from 0.59 pg/mL to 0.27 pg/mL, and from 37.96 pg/mL to 21.09 pg/mL, respectively. In the case of the omeprazole group, the level of TNF-α, IL-1β, and IL-6 decreased to 4.38 pg/mL, 0.27 pg/mL, and 18.58 pg/mL, respectively. The expression of PPAR-γ protein in the high-dose fisetin and omeprazole groups was about 1.5 times higher than that in the model group. Compared with the model group, the expression of NF-κB protein reduced to 0.34 level and 0.47 level in the omeprazole and high-dose fisetin groups, respectively. Compared with the model group, the expression of p38-MAPK protein reduced to 0.55 level and 0.68 level in the omeprazole and high-dose fisetin groups, respectively. Conclusion: Fisetin might relieve the symptoms of ethanol-induced gastric ulcer in rats through the regulation of NF-κB pathway.
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Rehmannia radix extract ameliorates imiquimod-induced psoriasis-like skin inflammation in a mouse model via the janus-kinase signal transducer and activator of transcription pathway p. 613

DOI:10.4103/pm.pm_218_19  
Background: Rehmannia glutinosa, commonly known as Dihuang, is a medicinal herb widely used in the formulations of traditional Chinese medicine. Radix or the root of Rehmannia is a vital ingredient used in various oriental medicines such as Jihwangeumja, Saenghyeoryunbuem, and Gyeongokgo. Hence, in the present study, we evaluated the potency of Rehmannia radix (RR) extract to ameliorate psoriasis like in vivo model induced by imiquimod (IMQ). Materials and Methods: Histopathological and histomorphometrical analysis was performed to assess the effect on RR extract against the psoriasis induction. To estimate whether the RR extract pretreatment inhibits the induction of psoriasis, the oxidative stress and antioxidants status were assessed. Further, to prove ameliorative effect of RR extract against psoriasis induction, the levels of inflammatory cytokines were estimated. Immunoblotting analyses of Janus kinase/signal transducer and activator of transcription (JAK/STAT) protein were estimated to confirm the molecular mechanism of RR extract against psoriasis induction. Results: RR extract effectively scavenged the free radicals, increased the antioxidant status, and decreased the inflammatory cytokines induced by IMQ. It also inhibits the expression of phosphorylated JAK/STAT protein, thereby preventing the dermis from inflammation. Histological analysis of psoriasis-induced and RR-pretreated mice' skin tissue authentically proves the inhibitory effect of RR extract against psoriasis induction. Conclusion: Our overall results suggest that RR extracts possess antipsoriatic property and can be subjected to human trial in the future as antipsoriatic drug.
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Naringenin reduced migration in osteosarcoma cells through downregulation of matrix metalloproteinase-2 and matrix metalloproteinase-9 and snail p. 620

DOI:10.4103/pm.pm_31_20  
Background: Osteosarcoma is one of the most malignant cancers in children. Naringenin exhibits several cellular functions. Objectives: In this study, we investigate the effects of naringenin on osteosarcoma. Materials and Methods: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was performed to detect the cell viability. Zymography assay and transwell assay were used to measure the matrix metalloproteinase (MMP) activity and migration ability. Western blot analysis was used to determine the expression of migration-related proteins. Results: No overt alternation of cytotoxicity in response to different concentrations of naringenin for 24 h was found by MTT assay. MMP-2 and MMP-9 activities and expression were significantly repressed by naringenin in a dose-dependent manner, as evidenced by zymography and Western blot analysis. Naringenin dose dependently reduced the expression of mesenchymal marker Snail. Conclusion: These results indicate that naringenin exhibited antimigration property through inhibition of MMP-2 and MMP-9 and downregulation of Snail expression. Thus, naringenin may be a potential inhibitor of metastasis of osteosarcoma.
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A special aromatic Chrysanthemum breed with high content of thujone p. 625

DOI:10.4103/pm.pm_382_19  
Background: A cultivar of Chrysanthemum, whose whole plant has strong and special fragrance, was found in Jiugong Mountain area, Xianning City, Hubei Province. Objective: The objective of the study was to determine taxonomic status of the cultivar in genus Dendranthema. Materials and Methods: The differences of morphological characteristics and volatile oil components between this cultivar and Dendranthema indicum, D. indicum var. aromaticum, and Dendranthema molifolium cv. “Jinju” were compared. Results: The cultivar was found to be a Chrysanthemum breed with unisexual flowers. The volatile oil content was 0.25% in its stem and leave of the breed and about 1.81% in its inflorescence which was much higher than that of other 3 species. There are 17 common components among volatile oils in inflorescence of the four Chrysanthemum species, but the thujone content of this cultivar is much higher than other three species. Conclusions: The cultivar is a new, cultivated Chrysanthemum breed, rich in volatile oil and thujone content, and named as Dendranthema morifolium (Ramat.) Tzvel. cv. “Jiugong Xiangju.”
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Effect and mechanism of ginsenoside Rg1 on synaptic plasticity of oxygen-glucose deprivation/reoxygenation-induced neuronal injury p. 630

DOI:10.4103/pm.pm_541_19  
Background: Ginsenoside Rg1 is the primary bioactive component of ginseng, which is a famous traditional Chinese medicine used to treat ischemic cardiovascular and cerebrovascular diseases. It has demonstrated considerable protective effects in neurons injured by ischemia/reperfusion both in in vitro and in vivoconditions. However, the effect and mechanism of action of ginsenoside Rg1 on the neural synaptic plasticity injured by ischemia/reperfusion have not yet been clarified. Objective: In this study, we aim to establish the model of oxygen–glucose deprivation/reoxygenation (OGD/R)-injured primary cortical neurons to mimic ischemia/reperfusion injury and investigate the mechanisms of action of ginsenoside Rg1 on the neural synaptic plasticity. Materials and Methods: Protective effects of ginsenoside Rg1 on neurons after OGD/R injury were measured by cell counting kit-8, lactate dehydrogenase, and apoptosis assay. The mRNA expression and activity of growth associated protein (GAP)-43, microtubule-associated protein (MAP)-2, Tau, insulin like growth factors (IGF)-1, brain derived neurotrophic factor (BDNF), and vascular endothelial growth facto r (VEGF) in neurons after OGD/R injury were measured by real-time polymerase chain reaction (RT-PCR) or enzyme-linked immunosorbent assay. The expression of apoptosis-related genes and cytosolic Ca2+ levels in neurons were determined via RT-PCR or Rhod-2 fluorescence staining. Results: According to our results, ginsenoside Rg1 protected the neurons and promoted axonal regeneration and neuronal remodeling after OGD/R injury; increased the expression of GAP-43, MAP-2, Tau, IGF-1, BDNF, VEGF, and Bcl-2 and inhibited the expression of Bax; and decreased the intracellular Ca2+ overload in OGD/R-injured neurons. Conclusion: Ginsenoside Rg1 can promote axonal regeneration and neuronal remodeling after OGD/R injury by upregulating the expression of synaptic remodeling proteins and endogenous neurotrophic factors, inhibiting intracellular Ca2+ overload and regulating the expression of apoptotic genes.
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Sagittaria sagittifolia polysaccharide, a chinese herbal extract, protects against isoniazid- and rifampicin-induced hepatotoxicity in in-vitro model p. 637

DOI:10.4103/pm.pm_542_19  
Background: Isoniazid (INH) and rifampicin (RFP) are first-line antituberculosis drugs; however, one of the most common adverse effects of their use is hepatotoxicity. Sagittaria sagittifolia polysaccharide (SSP) is the primary component of S. sagittifolia. Our previous research has confirmed the protective effect of SSP against INH and RFP-induced hepatic injury in in vivo model system. Objectives: In this study, we aimed to investigate the protective effect and the underlying mechanisms of SSP in cultured HepG2 cells after induction with INH and RFP. Materials and Methods: The study was designed as follows: normal (without any treatment), model (injury induced by INH + RFP), and SSP group (treated with SSP followed by INH + RFP). Cell viability, catalase (CAT), superoxide dismutase (SOD), and glutathione (GSH), along with malondialdehyde (MDA), as well as alanine aminotransferase, aspartate transaminase, and lactate dehydrogenase, were assessed by corresponding kits. Real-time polymerase chain reaction and Western blot were used to evaluate the mRNA and protein expression of Bcl-2, Bax, Nrf2, and Keap1. Results: SSP had a potential protective effect as it increased the cell viability and reduced the intracellular levels of hepatic injury markers. SSP reduced the intracellular content of MDA and increased the activity of SOD, CAT, and GSH. It is noteworthy that the optimum protective effect was observed with a 0.5 mg/mL SSP at 24 h after treatment. In addition, the protection appears to be associated with its activation of nuclear factor E2-related factor 2 signaling pathway. Conclusion: SSP protected against INH + RFP-induced hepatotoxicity in HepG2 cells and this protection might via Nrf2 signaling pathway.
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Methanolic extracts of Capparis ecuadorica iltis inhibit the inflammatory response in lipopolysaccharide-stimulated RAW 264.7 macrophage cells p. 644

DOI:10.4103/pm.pm_464_19  
Background: Some species of the caper family are known to possess antibacterial, antioxidant, anti-inflammatory, immunomodulatory, and antiviral properties. However, to date, the therapeutic effects of Capparis ecuadorica Iltis (Capparis L.) have not been studied. Objectives: In this study, we investigated the anti-inflammatory activity of a methanolic extract of C. ecuadorica leaves (MCE) in macrophages. Materials and Methods: Anti-inflammatory responses and mechanisms were assessed in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells after pretreatment with MCE. Results: In the MCE + LPS-treated group, the relative mRNA levels of pro-inflammatory cytokines (tumor necrosis factor (TNF)-α and interleukin (IL)-1β, IL-6) were downregulated and protein levels of IL-6 were also decreased as compared to the vehicle + LPS-treated group. Furthermore, the MCE + LPS-treated group showed reduced levels of reactive oxygen species (ROS) production as compared to the vehicle + LPS-treated group; detection of nitrite concentration revealed nitric oxide (NO) to be the reduced ROS. The expression levels of inducible nitric oxide synthase (iNOS)/cyclooxygenase-2 (COX-2) mRNA and the level of phosphorylation of IκBα were decreased in the MCE + LPS-treated group. In addition, MCE + LPS-treated group showed reduced levels of phosphorylation of MAP kinase in comparison to the vehicle + LPS-treated group. Interestingly, MCE also inhibited other inflammatory mechanisms, namely, endoplasmic reticulum (ER) stress and autophagy. Conclusion: These results indicate that MCE inhibits inflammatory responses through the inhibition of inflammatory cytokines and NO production, iNOS/COX-2 expression and nuclear factor-kappa B (NF-κB) activation, MAPK inhibition, as well as regulation of ER stress and autophagy in LPS-stimulated RAW 264.7 cells.
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Distinguishing the rhizomes of Atractylodes japonica, Atractylodes chinensis, and Atractylodes lancea by comprehensive two-dimensional gas chromatography coupled with mass spectrometry combined with multivariate data analysis p. 654

DOI:10.4103/pm.pm_33_20  
Background: In clinical practice, the species of Atractylodes are difficult to identify based on their morphological and chemical features which often leads to confusion. In addition, the composition of volatile components may influence the clinical efficacy of rhizomes of Atractylodes. Materials and Methods: In this study, a comprehensive two-dimensional gas chromatography with mass spectrometry coupled with multivariate data analysis was employed to investigate the differences in the volatile components of the rhizomes of three species of Atractylodes, namely Atractylodes lancea (Thunb.) DC, Atractylodes japonica Koidz. et Kitam, and Atractylodes chinensis (DC.) Koidz. Results: A total of 119 compounds were tentatively identified and confirmed based on the NIST database. Thirty-three samples were well distinguished and the results of two different analytical methods using principal component analysis and partial least-squares discriminant analysis were in satisfactory agreement with one-way analysis of variance. Atractylodin and β-eudesmol can be used to reveal the chemical differentiation and distinguish different species of Atractylodes. Conclusion: The results may provide a reliable reference to quality control and product grade of rhizomes of Atractylodes.
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Optimization of ultrasound-assisted enzymatic extraction and antioxidant activity of polysaccharide from radix Morindae officinalis by response surface methodology p. 662

DOI:10.4103/pm.pm_444_19  
Objectives: In this study, we aimed to establish an efficient method for the extraction of radix Morindae officinalispolysaccharide (MOP) by applying ultrasonic technology and by optimizing the parameters through response surface methodology (RSM) based on the central composite design. Materials and Methods: Ultrasound-assisted enzymatic extraction (UAEE) was performed to extract the MOP. We applied an orthogonal array design to optimize the concentration of enzymes (cellulose, pectinase, and papain). The extraction parameters were optimized based on the RSM technique. Furthermore, the effects of ultrasound-assisted enzymatic treatment on 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical-scavenging activity by the MOP was optimized using the RSM technique. Results: According to our results, 1.0% cellulase, 1.5% pectinase, and 1.0% papain were the optimum concentrations of the enzyme (calculated based on the percentage dry weight of radix Morindae officinalis powder). The optimum conditions of extraction were as follows: 21 mL/g solvent–solid ratio, 280 W, pH of 5.3, and temperature of 50°C. Under these conditions, the yield of MOPs and DPPH-scavenging activity of MOP were 23.68% ± 0.52% (n = 3) and 117.26 ± 2.73 mg Vitamin C/100 g dry weight (n = 3), respectively. The overall desirability was 1.03 ± 0.01 (n = 3). Conclusion: UAEE was effective in the extraction of MOP, and RSM technique was adequate to design and optimize the extraction parameters.
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Content determination and anti-fatigue effect of the purified anthocyanin from purple Daucus carota p. 670

DOI:10.4103/pm.pm_452_19  
Background: The content and anti-fatigue effect of the purified anthocyanin from purple Daucus carota(PAPD) were studied. Materials and Methods: The content of total anthocyanins in PAPD was determined by the pH trial-difference method, and the content of cyanidin-3-o-glucoside in PAPD was determined by high-performance liquid chromatography (HPLC) method. Hundred ICR mice were randomly divided into five groups with 20 mice in each group: Negative control (NC) group, positive control (PC, red bull drink of 1.0 g/kg BW) group, PAPD dose group of low, middle, high (Low, 0.2 g/kg; Mid, 0.4 g/kg; Hig, 0.8 g/kg). Mice were intragastric injected continuously for 4 weeks; once a day, the NC group received an equal volume of saline, the PC group was given 1.0 g/kg BW red bull drink. After the last administration, the time of weight-bearing exhaustion swimming of 10 mice in each group was determined, the contents of lactic acid and urea nitrogen in the serum and glycogen in the liver and muscle tissues after 30 min of no-weight-bearing swimming of ten mice in each group were determined and the weight of mice during the experiment was also determined and the pathological analysis of the liver and kidney tissues in mice was conducted. Results: The content of total anthocyanin in PAPD was determined by the pH-differential method to be 132.46 mg/g DW, which was converted into fresh weight as 1.146 mg/g. The content of cyanidin-3-O-glucoside in PAPD was determined by HPLC, which was 31.45 mg/g DW, accounting for 21.37% of the total anthocyanin. Compared with the NC group, the exhausted swimming time of the PC group and PAPD group was significantly prolonged (P < 0.05), the contents of muscle glycogen and liver glycogen in the tissues of mice were significantly increased (P < 0.05), the contents of lactic acid and urea nitrogen in the serum of mice were significantly reduced (P < 0.05), and the lesions of the liver and kidney tissues were enhanced to some extent. Conclusion: The content of anthocyanin is rich in PAPD and has an observable anti-fatigue effect; its potential mechanism could improve the metabolism of sugar and lipid in muscle and liver tissues, reduce the oxidative damage of cells, improve the adaptability and endurance of the body to strenuous exercise, and enhance the immunity and antioxidant ability of cells, to play an anti-fatigue role.
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Glabridin downregulates lipopolysaccharide-induced oxidative stress and neuroinflammation in BV-2 microglial cells via suppression of nuclear factor-κB signaling pathway p. 675

DOI:10.4103/pm.pm_497_19  
Background: Microglia initially undergoes chronic activation in response to the damages caused by stressful stimuli, such as bacterial infection and hypoxia. Inflammatory responses, as well as oxidative stress, perform crucial roles in the process of neuroinflammation, which results in brain damage. Glabridin is a natural organic compound with extensive beneficial properties such as anti-inflammatory and antioxidant properties. To the best of our knowledge, there are no studies conducted on the anti-neuroinflammatory activity of glabridin. Therefore, in this study, we aimed to investigate the potency of glabridin against the expression of lipopolysaccharide (LPS)-stimulated BV-2 cells. Materials and Methods: BV-2 cells were preincubated with glabridin followed by the LPS challenge. Subsequently, the cellular status of nitric oxide (NO), reactive oxygen species (ROS), prostaglandin E2(PGE2), and pro-inflammatory modulators (interleukin [IL]-1β and IL-6) were investigated and related signaling pathways were inspected via blotting assay. Results: Our results indicate that glabridin appreciably alleviated the LPS-induced accretion of inducible-NO synthase (iNOS), PGE2, IL-1β, and IL-6. Moreover, it noticeably allayed the NO/iNOS, PGE2/cyclooxygenase-2 protein statuses, and pro-inflammatory cytokine (tumor necrosis factor-α) on LPS-induced microglia. We also found that LPS severely increased the phosphorylation of Inhibitory kappa B kinases (IKKs), IκBα, p65, and nuclear factor (NF)-κB. Although glabridin supplementation suppressed the phosphorylation of the aforementioned molecules, LPS remarkable caused the nuclear interchange of NF-κBp65. Conclusion: Glabridin alleviates LPS-induced neuroinflammation in BV-2 cells by suppressing the accumulation of ROS and cell death and by inhibiting the pro-inflammatory responses via NF-κB-dependent mechanisms. According to our results, glabridin may be beneficial in neuroinflammation-related neurodegenerative disorders.
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Fucoxanthin modulates the development of 7, 12-dimethyl benz (a) anthracene-induced skin carcinogenesis in swiss albino mice in vivo p. 681

DOI:10.4103/pm.pm_292_19  
Background: Fucoxanthin (Fx), an distinctive carotenoid occurs on brown seaweed, contains several benefits including anti-cancer effects. To investigate the chemopreventive effectual of Fx on 7,12-dimethylbenz[a] anthracene (DMBA)-alone skin tumor development in Swiss albino mice. Materials and Methods: The skin sarcoma being provoked on the hairless flipside of the mice's skin, twice weekly for 8 weeks through challenging through DMBA (25 μg on 0.1 ml acetone/mice). Thereafter, the mice were oral supplementation with 50 mg/kg body weight (BW), Fx for 25th week a frequency of three times/week. Tumor size, change in BW, the cumulative quality of papillomas and some oxidative stress-related parameters were measured. Results: Orally administered Fx were notably increased BW, delayed tumor incidence with no abnormal pathology in DMBA-induced skin tumor mice. Fx effectively modulates the level of xenobiotic enzymes, brought back to the statuses of lipid peroxidation (LPO) enzymes and increased antioxidant enzymes status in squamous cell carcinomas (SCC). A considerable decrease in the protein expression of proinflammatory markers that is interleukin (IL) 1-6, tumor necrosis factor-alpha (TNF-α), and IL-β and restored the expression status of inflammatory regulators such as nuclear factor-kappa B (NF-κB), cyclooxygenase-2 (COX-2), and inducible nitric oxide synthase (iNOS) on serum. These findings were supported through histopathological examinations. Hence, it was clear that Fx possessed a good anticancer activity against skin cancer. Conclusion: The chemopreventive prospective of Fx was eventually owing to its altering capacity on the levels of proinflammatory (IL 1-6, TNF-α, and IL-β) cytokines, inflammatory (NF-κB, COX-2, and iNOS) markers, LPO, antioxidants, and toxin-eliminating mediators in DMBA-provoked skin SCC. Thus, it can be concluded that the FX indicates the antitumor potential on DMBA challenged skin tumor proliferation model on mice.
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