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April-June 2017
Volume 13 | Issue 50
Page Nos. 203-337

Online since Tuesday, April 18, 2017

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ORIGINAL ARTICLES  

Assessment of mexican arnica (Heterotheca inuloides Cass) and rosemary (Rosmarinus officinalis) extracts on dopamine and selected biomarkers of oxidative stress in stomach and brain of Salmonella typhimurium infected rats p. 203
David Calderķn Guzmān, Maribel Ortiz Herrera, Norma Osnaya Brizuela, Gerardo Barragàn Mejía, Ernestina Hernàndez García, Hugo Juàrez Olguín, Armando Valenzuela Peraza, Norma Labra Ruíz, Daniel Santamaría Del Angel
DOI:10.4103/0973-1296.204553  
Background: The effects of some natural products on dopamine (DA) and 5-hydroxyindole acetic acid (5-HIAA) in brain of infected models are still unclear. Objective: The purpose of this study was to measure the effect of Mexican arnica/rosemary (MAR) water extract and oseltamivir on both biogenic amines and some oxidative biomarkers in the brain and stomach of young rats under infection condition. Methods: Female Wistar rats (weight 80 g) in the presence of MAR or absence (no-MAR) were treated as follows: group 1, buffer solution (controls); oseltamivir (100 mg/kg), group 2; culture of Salmonella typhimurium (S.Typh) (1 × 106 colony-forming units/rat) group 3; oseltamivir (100 mg/kg) + S.Typh (same dose) group 4. Drug and extracts were administered intraperitoneally every 24 h for 5 days, and S.Typh was given orally on days 1 and 3. On the fifth day, blood was collected to measure glucose and hemoglobin. The brains and stomachs were obtained to measure levels of DA, 5-HIAA, glutathione (GSH), TBARS, H2O2, and total ATPase activity using validated methods. Results: DA levels increased in MAR group treated with oseltamivir alone but decreased in no-MAR group treated with oseltamivir plus S.Typh. 5-HIAA, GSH, and H2O2 decreased in this last group, and ATPase activity increased in MAR group treated with oseltamivir plus S.Typh. TBARS (lipid peroxidation) increased in MAR group that received oseltamivir alone. Most of the biomarkers were not altered significantly in the stomach. Conclusion: MAR extract alters DA and metabolism of 5-HIAA in the brain of young animals infected. Antioxidant capacity may be involved in these effects. Abbreviations used: AS: Automated system, ATP: Adenosine triphosphate, CNS: Central nervous system, CFU: Colony-forming unit, DA: Dopamine EDTA: Ethylenediaminetetraacetic acid, 5-HIAA: écido 5-hidroxindolacético (serotonina), GABA: γ-aminobutyric acid, GSH: Glutathione, H2O2: Hidrogen peroxide, HCLO4: Perchloric acid, iNOS: Inducible nitric oxide synthase, LPS: Lipopolysaccharides, MAR: Arnica/ Rosemary, NaCl: Sodium Chloride, NOGSH: nitrosoglutathione,NOS: Nitric oxide, OPT: Ortho-phtaldialdehyde, Pbs: Phosphate buffered saline, pH: potential of Hydrogen, Pi: Inorganic phosphate, ROS: Reactive oxygen species, RNSs: Reactive nitrogen species Tba: Thiobarbaturic acid, TBARS: Thiobarbituric aid reactive, Tca: Trichloroacetic, Tris-HCL: Tris hydrochloride, TSA: Trypticasein Soya Agar
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Multipathway integrated adjustment mechanism of Glycyrrhiza triterpenes curing gastric ulcer in rats p. 209
Ying Wang, Shuai Wang, Yongrui Bao, Tianjiao Li, Xin Chang, Guanlin Yang, Xiansheng Meng
DOI:10.4103/0973-1296.204550  
Background: Gastric ulcer is a common chronic disease in human digestive system, which is difficult to cure, easy to relapse, and endangers human health seriously. Compared with western medicine, traditional Chinese medicine has a unique advantage in improving the general situation, stablizing medical condition, and with little side effects. Glycyrrhiza known as “king of all the medicine”, has a range of pharmacological activities and is commonly used in a variety of proprietary Chinese medicines and formulations. Objective: On the basis of explicit antiulcer effect of Glycyrrhiza triterpenes, the molecular mechanisms of its therapeutic effect on acetic acid induced gastric ulcer in rats were explored. Materials and Methods: Acetic acid induced gastric ulcer model in rats was established to evaluate the curing effect of G. triterpenes and all of the rats were randomised into six groups: Control group, model group, omeprazole group (0.8 mg/mL), triterpenes high dose group (378.0 mg/mL), triterpenes middle dose group (126.0 mg/mL), and triterpenes low dose group (42.0 mg/mL). All rats in groups were orally administered the active group solution 1.5 mL once daily (model and control groups with saline) for 7 days. HPLC-TOF-MS analysis method was performed to obtain the plasma metabolites spectrums of control group, model group, triterpenes high, middle and low dose groups. Results: A total of 11 differential endogenous metabolites related to the therapeutic effect of G. triterpenes were identified, including tryptophan, phingosine-1-phosphate, pantothenic acid, and so on, among which tryptophan and phingosine-1-phosphate are related with the calcium signaling pathway and arachidonic acid (AA) metabolism. At the same time, in order to verify the above results, quantitative real time polymerase chain reaction were performed to evaluate the expression of H+-K+-ATPase alpha mRNA and phospholipase a 2 mRNA in relational signaling pathways. Combined with statistical analysis of plasma metabolic spectrum and gene expression in tissue, it is suggested that G. triterpenes has antiulcer effect on gastric ulcer in rats. Conclusion: G. triterpenes has a certain regulating effect on the metabolism of tryptophan, AA, sphingosine, and other endogenous metabolites, and we speculated that the antiulcer potential of G. triterpenes can be primarily attributed to its inhibiting gastric acid secretion, reducing the release of inflammatory mediators, and protecting gastric mucosa effects to prevent the further development of gastric ulcer. Abbreviations used: HP: Helicobacter pylori, ECL: enterochromaffinlike,TCM: Traditional Chinese medicine; HPLC: High Performance Liquid Chromatography, HPLC/MS: High Performance Liquid chromatography Mass Spectrometry, HPLC-TOF-MS: High Performance Liquid Chromatography and Tof Mass Spectrometry, SD: Sprague Dawley, PCDL: Personal Compound Database and Library, MPP: Mass Profiler Professiona; PCA: principal component analysis, RT-PCR: real time polymerase chain reaction, PGE 2: Prostaglandin E2, COX1: cyclooxygenase 1 S1P: Sphingosine-1-phosphate, AA: Arachidonic acid, 5-HT: 5- hydroxytryptamine.
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Anti-inflammatory effects of KOTMIN13: A mixed herbal medicine in LPS-stimulated RAW 264.7 cells and mouse edema models p. 216
Eujin Lee, Sun-Gun Kim, Na-Young Park, Hyo-Hyun Park, Kyu-Tae Jeong, Jongkeun Choi, In-Hae Lee, Hwadong Lee, Eunkyung Lee
DOI:10.4103/0973-1296.204548  
Background: A Korean herbal medicine, KOTMIN13, composed of Inula japonica Thunberg, Trichosanthes kirilowii Maximowicz var. japonica kitamura, Peucedanum praeruptorum Dunn, and Allium macrostemon Bge, has been used for anti-allergic and anti-asthmatic treatment in oriental clinics, but its activity has not been investigated. Materials and Methods: To evaluate the anti-inflammatory activity of KOTMIN13 for in vitro study, LPS-stimulated RAW 264.7 cells were used to induce the production and expression of inflammatory mediators and its mechanisms. 12-O-Tetradecanoylphorobol-13 aceate (TPA)-induced ear edema and carrageenan-induced paw edema models were also used to evaluate the effect of KOTMIN13 on acute inflammation in vivo. Results: KOTMIN13 reduced the release of inflammatory mediators [nitric oxide, prostaglandin E2, interleukin (IL)-1β, and IL-6] and the protein expression of inducible nitric oxide synthase and cyclooxygenase-2 in LPS-stimulated RAW 264.7 cells. Mechanism studies showed the attenuation of LPS-induced NF-κB activation by KOTMIN13 via IκBα degradation abrogation and a subsequent decrease in nuclear p65 levels. Activation of mitogen-activated protein kinases (ERK, JNK, and p38) was also suppressed. Furthermore, KOTMIN13 ameliorated the development of TPA-induced ear edema and carrageenan-induced paw edema in acute inflammatory edema mouse models. Conclusion: Our study demonstrates that KOTMIN13 inhibits inflammatory mediators through the inhibitions of NF-κB and MAPK activities in LPS-induced RAW 264.7 cells, as well as acute inflammation in edema models, indicating that KOTMIN13 is an effective suppressor for anti-inflammatory activities. Abbreviations used: NO: nitric oxide; PGE2: prostaglandin E2; iNOS: inducible NO synthase; COX-2: cyclooxygenase-2; TNF-α: tumor necrosis factor-α; IL: interleukin; NF-κB: nuclear factor kappaB; MAPK: mitogen-activated protein kinases; ERK: extracellular signal regulated kinase; JNK: c-jun N terminal kinase; TPA: 12-O-tetradecanoylphorbol-13-acetate
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Flavonoids isolated from the flowers of Limonium bicolor and their in vitro antitumor evaluation p. 222
Jian Chen, Jiehui Teng, Li Ma, Haiying Tong, Bingru Ren, Linshan Wang, Weilin Li
DOI:10.4103/0973-1296.204566  
Background: Limonium bicolor, a halophytic species, can grow in saline or saline-alkali soil, is well known as a traditional Chinese medicine. Recently it attracted much attention for its treatment for cancer. Objective: The present study was performed to evaluate this species from the phytochemical standpoint and the possible relationship between the antitumor activity and its natural products. Materials and methods: The chemical constituents from the flowers of L. bicolor were investigated through bioassay-guided fractionation and isolation. All the individual compounds were characterized by spectroscopic analysis and their potential antitumor activity was tested against three different human tumor cell lines by MTT assays. Results: The EtOAc extract was proven as the most potent fraction and further fractionation led to the isolation of 15 natural flavonoids, which were characterized as luteolin (1), acacetin (2), quercetin (3), isorhamnetin (4), kaempferol (5), eriodictyol (6), kaempferol-3-O-α-L-rhamnoside (7), kaempferol-3-O-β-D-glucoside (8), quercetin-3-O-α-L-rhamnoside (9), quercetin-3-O-β-D-glucoside (10), quercetin-3-O-β-D-galactoside (11), myricetin-3-O-α-L-rhamnoside (12), kaempferol-3-O-(6″-O-galloyl)-β-D-glucoside (13), hesperidin (14) and rutin (15). The biotesting results demonstrated that both compounds 1 and 3 showed good cytotoxicity against human colon cancer cells (LOVO). Compound 5 exhibited relative greater growth inhibition against both human breast cancer cells (MCF-7) and osteosarcoma cell lines (U2-OS) at the concentration of 100 μg/mL. Conclusion: On the basis of these findings, the flavonoids were deduced to be potentially responsible for the antitumor activity of L. bicolor. The preliminary structure–activity relationship analysis suggests that the 3-O-glycosylation moiety in natural flavonoids was not essential for the antiproliferative activity on LOVO and U2-OS cells. Abbreviation used: MTT: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, EtOAc: Ethyl acetate; LOVO: human colon cancer; MCF-7: human breast, cancer; U2-OS: human osteosarcoma; 5-FU: 5-Fluorouracil; DMSO: dimethyl sulfoxide, NMR: nuclear magnetic resonance; HR-ESI-MS: high resolution electrospray ionization mass chromatography, HPLC: high performance liquid chromatography, EtOH: ethanol; n-BuOH: n-butanol; CC: column chromatography, TLC: thin layer chromatography; PBS: phosphate-buffered saline.
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Betulinic acid from Zizyphus Joazeiro bark using focused microwave-assisted extraction and response surface methodology p. 226
Fernanda Carolina Sousa Fonseca, Letícia Caribé Batista Reis, Jener David Gonçalves dos Santos, Carla Rodrigues Cardoso Branco, Sergio Luis da Costa Ferreira, Jorge Mauricio David, Alexsandro Branco
DOI:10.4103/0973-1296.204565  
Background: The effect of the extraction time (min) and temperature (°C) on the yield of betulinic acid (BA) from Zizyphus joazeiro barks using focused microwave-assisted extraction was investigated. Materials and Methods: The ethyl acetate was used as extractor solvent because it was shown to provide a betulinic acid-clean extract. A full two-level statistical factorial design was applied to determine the important effects and interactions of these independent variables upon the yield of BA. Results: The conditions that produced the highest yield of BA were at temperature of 70°C and an extraction time of 15 min (3.33 mg per gram of plant). Conclusion: The BA has drawn attention due to its use as a raw material in the synthesis of active compounds against the Human Immunodeficiency Virus (HIV). Abbreviation used: BA: Betulinic acid; FMAE: Focused microwave assisted extraction; HPLC: High-performance liquid chromatography; RSD: Relative standard deviations.
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Content determination of phenylpropanoids and enhancing exercise ability of effective fractions in pedicularis densispica p. 230
Hongbiao Chu, Zhihua Zhang, Dong Chen, Xi Wang, Qilong Tu
DOI:10.4103/0973-1296.204567  
Background: Most researches were focused on chemical constituents and bioactivities of Pedicularis. However, there were a few reports on simultaneous determination of the series phenylpropanoids compounds in Pedicularis by High Performance Liquid Chromatography (HPLC). Objective: To establish an HPLC method for simultaneous determination of salidroside, verbascoside, iso-verbascoside, leucoseptoside A, jionoside D and martynoside in Pedicularis densispica (PD), and to assess the enhancing exercise ability of effective fractions of phenylpropanoids (EFP). Materials and Methods: The separation was performed on C18 column with step-wise gradient elution with water (A)-methanol (B) as the mobile phase at a flow rate of 1.0 mL/min, with detection wavelength at 275 nm (0–4 min) and 330 nm (4–40 min). The EFP were obtained from extracts of PD by resin gradient dilution. The enhancing exercise ability of EFP was exerted in exhaustive swimming and anoxia endurance tests in vivo. Results: The contents of six marker compounds had good linear relationship in the ranges of 2.10–8.40, 13.60–54.40, 0.93–3.72, 0.53–2.12, 1.50–6.00, 0.37–1.28, respectively, and the average recoveries of the six phenylpropanoids were all in the range of 98–103%. Total contents of phenylpropanoids in EFP were more than 60%. Three medicine groups of exhaustive swimming and anoxia endurance time were higher than those of the water group. Conclusion: The analytical method is reliable, simple and accurate, and can be used for the comprehensive quality control of PD. This experiment suggests that PD has the effect of promoting the recovery and elimination of fatigue and improving the exercise capacity. Abbreviation used: PD: Pedicularis densispica, EFP: Effective fractions of phenylpropanoids, DAD: Diode array detector, HPLC: High performance liquid chromatography, LOD: Limits of detection, LOQ: Limits of quantification, RSD: Relative standard deviation, BV: Bed volumes
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Evaluation of herb–drug interactions of Hovenia dulcis fruit extracts p. 236
Jong Suk Park, Shaheed Ur Rehman, In Sook Kim, Min Sun Choi, Chun-Soo Na, Hye Hyun Yoo
DOI:10.4103/0973-1296.204552  
Background: Hovenia dulcis (Rhamnaceae) fruits are popularly used as herbal medicines or dietary supplements in Asian countries due to functions such as liver protection and detoxification from alcohol poisoning. Accordingly, it is very likely for dietary supplemental products, including H. dulcis fruit extracts, to be taken with prescription drugs. Objective: In this study, possible food–drug interactions involving H. dulcis fruit extracts were evaluated based on the inhibition of cytochrome P450 (CYP) enzyme activity. Material and Methods: The water extract of H. dulcis fruit extracts was incubated in human liver microsomes with CYP-specific substrates. The formation of the CYP-specific metabolites was measured using liquid chromatography-tandem mass spectrometry. Results: H. dulcis fruit extracts showed negligible effects on seven CYP isozyme activities at all concentrations tested. Conclusion: This result suggests that H. dulcis fruit extracts may have minimal pharmacokinetic interactions with coadministered drugs through the modulation of CYP enzymes. Abbreviations Used: CYP: cytochrome P450 enzymes, HPLC: High performance liquid chromatography, LC-MS/MS : liquid chromatography-tandem mass spectrometry, MRM: multiple-reaction monitoring
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Antiprotozoal, antibacterial and antidiarrheal properties from the flowers of Chiranthodendron pentadactylon and isolated flavonoids p. 240
Fernando Calzada, Teresa Juárez, Normand García-Hernández, Miguel Valdes, Oscar Ávila, Lilian Yepez Mulia, Claudia Velázquez
DOI:10.4103/0973-1296.204564  
Background: Chiranthodendron pentadactylon Larreat. (Sterculiaceae) is a Mexican plant used in traditional medicine for the treatment of heart disease symptoms and infectious diarrhea. Objective: To evaluate in vitro antiprotozoal and antibacterial activities and in vivo antidiarrheal activity from the flowers of C. pentadactylon using the extract, fractions, and major isolated flavonoids. Materials and methods: Bioassay-guided fractionation of the methanol extract of C. pentadactylon (MECP) led to the isolation of five flavonoids, tiliroside, astragalin, isoquercitrin, (+)-catechin, and (-)-epicatechin. Antimicrobial activities were tested on two protozoa (Entamoeba histolytica and Giardia lamblia) and nine bacterial enteropathogens (two Escherichia coli strains, two Shigella sonnei strains, two Shigella flexneri strains, two Salmonella sp. strains, and Vibrio cholerae) isolated from feces of children with acute diarrhea or dysentery and resistant to chloramphenicol. Also, antidiarrheal activity was tested on cholera toxin-induced diarrhea in male Balb-c mice. Results: Epicatechin was the most potent antiamoebic and antigiardial compound with IC50 values of 1.9 μg/mL for E. histolytica and 1.6 μg/mL for G. lamblia; tiliroside showed moderate antiprotozoal activity against both protozoan. In contrast, in the antibacterial activity, tiliroside was the most potent compound on all microorganisms with minimum inhibitory concentration values less than 0.7 mg/mL. In the case of cholera toxin-induced diarrhea, epicatechin was the most potent flavonoid with IC50 of 14.7 mg/kg. Conclusion: Epicatechin and tiliroside were the flavonoids responsible for antimicrobial andantidiarrheal activities of C. pentadactylon. Its antiprotozoal, antibacterial, and antidiarrheal properties are in good agreement with the traditional medicinal use of C. pentadactylon for the treatment of infectious diarrhea. Abbreviations used: MECP: Methanol extract of C. pentadactylon
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Prevention mechanism of 2,3,5,4'-tetrahydroxy-stilbene-2-O-β-D-glucoside on lipid accumulation in steatosis hepatic L-02 cell p. 245
Pei Lin, Jian-Mei Lu, Yan-Fang Wang, Wen Gu, Rong-Hua Zhao, Jie Yu
DOI:10.4103/0973-1296.204563  
Aim: 2,3,5,4'-Tetrahydroxy-stilbene-2-O-β-d-glucoside (TSG), a natural stilbene, shows great activities in hepatic lipid regulation, especially for hepatic triglyceride lowering. However, information about its mechanisms on biosynthesis and degradation of triglyceride is still limited. This research pays close attention to clarify the mechanism of TSG on prevention of hepatic lipid accumulation. Materials and Methods: TSG was given to steatosis hepatocyte L-02 cell induced by fat emulsion incubation. The contents of free fatty acid, triglyceride, rate-controlling enzymes, and transcriptional regulatory factors, which play key role in biosynthesis and decomposition of triglyceride, were determined with or without TSG exposure. Results: TSG could reduce the free fatty acid material supply for the synthesis of endogenous triglyceride and it did so by reducing the expression of liver type fatty acid binding protein and fatty acid transport protein 4. TS Ginhibited the expression of sterol regulatory element-binding protein 1c, and then reduce the contents of acetyl-CoA carboxylase 1 and fatty acid synthase. Therefore,TSG prevented biosynthesis of triglyceride. Mean while, TSG also promoted the decomposition of triglyceride by the activation of peroxisome proliferators activator receptors alpha. Conclusion: TSG could effective intervene the accumulation of triglyceride in hepatic cell. Thus, TSG could be considered as a promising drug candidate in prevention and treatment of lipid metabolic disorders, especially nonalcoholic fatty liver disease. Abbreviations used: ACACA: Acetyl-CoA carboxylase 1, Apo-B100: Apo lipoprotein B100, FASN: Fatty acid synthase, FATP4: Fatty acid transport protein 4, FBS: Fetal bovine serum; FEN: Fenofibrate, FFA: Free fatty acid, L-FABP: Liver type fatty acid binding protein, LPL: Lipoprotein lipase, MTTP: Microsomal triglyceride transfer protein, NAFLD: Non-alcoholic fatty liver disease, PBS: Phosphate buffer saline, PPAR-α: Peroxisome proliferators activator receptors alpha, RPMI: Roswell Park Memorial Institute, SIM: Simvastatin, SREBF1c: Sterol regulatory element-binding protein 1c, TG: Triglyceride, TSG: 2, 3, 5, 4-tetrahydroxy-stilbene-2-O-β-Dglucoside, VLDL: Very low density lipoprotein
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Caspase-mediated apoptotic effects of Ebenus boissieri barbey extracts on human cervical cancer cell line hela p. 254
Ece Simsek, Nilufer Imir, Esra Arslan Aydemir, Ramazan Suleyman Gokturk, Erdem Yesilada, Kayahan Fiskin
DOI:10.4103/0973-1296.204560  
Background: Ebenus boissieri Barbey is an Antalya, Turkey-endemic plant belonging to Fabaceae family. The aerial parts and the roots of E. boissieri Barbey were used in this study. Objective: In the present study, we have examined the apoptotic effects of hydroalcoholic extracts of E. boissieri Barbey in human cervical cancer cell line HeLa. Materials and Methods: To determine the cytotoxic effect, cells were treated with various concentrations of extracts for 24, 48, and 72 h incubation periods. Cytotoxic effects were examined by Cell Titer 96 aqueous nonradioactive cell proliferation assay and the results were corrected by live/dead viability/cytotoxicity assay and trypan blue exclusion assay. Apoptotic effects were studied with multicaspase kit. Tumor necrosis factor-alpha (TNF-α) and interferon gamma (IFN-γ) release were also measured by enzyme-linked immunosorbent assay. Results: According to the results, E. boissieri Barbey extract caused significant increase in caspase levels. Thus, we suggest that the extract induces cells to undergo apoptosis. Especially, there was a sharp induction in caspase-3 activity. Levels of both TNF-α and IFN-γ in extract-treated groups were significantly and dose dependently exalted as compared to their relative controls. Conclusion: The effects of the extract on caspase-3, TNF-α, and IFN-γ levels mediate the plausible mechanism of apoptosis induction in HeLa. To the best of our knowledge, this is the first report indicating any pharmacological properties of E. Boissieri on HeLa cells. Abbreviations used: Tumor necrosis factor-alpha (TNF-α); Interferon gamma (IFN-γ); 3-(4, 5 dimethylthiazol-2-yl)-5-(3- carboxymethoxy-phenyl)-2-(4-sulfonyl)-2H-tetrazolium (MTS); Phosphate-Buffered Saline (PBS); Fetal Bovine Serum (FBS); para-Nitroanilin pNA; Enzyme-Linked ImmunoSorbent Assay (ELISA); Sodium Dodesyl sulphate –Polyacrilamide gel electrophoresis (SDS-PAGE); Tris-Buffered Saline (TBS); Hydocloric acid (HCl); Standart Error of Mean (SEM); National Cancer Institute (NCI); half maximal inhibitory concentration (IC50)
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Azorella compacta infusion activates human immune cells and scavenges free radicals in vitro p. 260
Lenka Tumová, Zuzana Dučaiová, José Cheel, Ivan Vokřál, Beatriz Sepúlveda, Doris Vokurková
DOI:10.4103/0973-1296.204558  
Background: Azorella compacta is traditionally used in the form of tea (infusion), in the Andean region of South America, to treat various chronic diseases. However, the health-promoting properties of this herbal tea have not yet been extensively explored. Materials and Methods: The free radical scavenging activity of A. compacta infusion (ACI) was evaluated by the 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical and superoxide anion radical assays. The activation of immune cells by ACI, as determined by cell surface cluster of differentiation 69 expression, was measured by flow cytometry. The qualitative polyphenolic composition of ACI was investigated by HPLC/PDA/ESI-MS, (High-performance liquid chromatography coupled with photodiode array detection and electrospray ionization - mass spectrometry) and the total content of polyphenols was estimated by spectrophotometric methods. Results: Eight polyphenols including chlorogenic acid, 6,8-di-C-hexosyl apigenin, isoorientin, orientin, dicaffeoylquinic acid, biochanin A-O-glucoside, biochanin A-O-(malonyl)-glucoside, and licoisoflavone A were tentatively identified in ACI. The total contents of phenols, flavonoids, and tannins in lyophilized ACI were 5.40 mg/100 mg ACI, 1.79 mg/100 mg ACI, and 1.76 mg/100 mg ACI, respectively. ACI, within the range of 25-400 µg/mL, scavenged DPPH and O2.– by 15-90% and 20-88%, respectively. The human natural killer (NK) cells were substantially activated by ACI, whereas T cells and granulocytes were slightly stimulated. Conclusion: Overall, the results demonstrate the free radical scavenging and immune-stimulating properties of ACI, and support, at least in part, its potential utilization as a functional herbal tea. for preventing chronic diseases and as a nonspecific immune stimulator during human immunosenescence. Abbreviations used: ESI: electrospray ionization, HPLC: high performance liquid chromatography, PDA: photodiode array detector, MS: mass spectrometry, MS/MS: tandem mass spectrometry, MW: molecular weight, m/z: mass-to-charge ratio, FITC: fluorescent isothiocyanate, PE: phycoerythrin
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Ultrasound-assisted extraction of ursolic acid from the flowers of Ixora coccinia linn (Rubiaceae) and antiproliferative activity of ursolic acid and synthesized derivatives p. 265
Silvana Amadeu Ferreira Alves Monteath, Maria Aparecida M Maciel, Raquel Garcia Vega, Heloisa de Mello, Carollina de Araújo Martins, Andressa Esteves-Souza, Cerli Rocha Gattass, Aurea Echevarria
DOI:10.4103/0973-1296.204557  
Background: Ixora coccinea Linn (Rubiaceae) is an evergreen shrub with bright scarlet colored flowers found in several tropical and subtropical countries. It is used as an ornamental and medicinal plant. Phytochemical studies revealed that its major special metabolites are triterpene acids, such as ursolic and oleanolic acid. Objective: To evaluate the isolation of ursolic acid (UA) (1) from methanol extracts of I. coccinea flowers through two methodologies, to prepare four derivatives, and to evaluate the cytotoxic effect against six cancer cell lines. Materials and Methods: The UA was isolated from vegetal material by percolation at room temperature and by ultrasound-assisted extraction. The preparation of derivatives was performed according to literature methods, and the cytotoxic effects were evaluated using the MTT (3,4,5-dimethylthiazol-2-yl-2,5-diphenyltetrazolium bromide) assay. Results: The most efficient extraction was achieved through ultrasound irradiation with a yield of 35% after KOH-impregnated silica in chromatography column. Furthermore, four derivatives (3, 5, 6, 7) of UA were prepared and evaluated, including 1, against two lung cancer (A549 and H460) and four leukemia (K562, Lucena, HL60, and Jurkat) cell lines. Generally, results showed that 1 and 7 were the most active compounds against the assayed cell lines. Also, the cytotoxic effects observed on terpenes 1 and 7 were higher when compared with cisplatin, used as positive control, with the exception of Jurkat cell line. Conclusion: The efficiency of such an alternative extraction method led to the principal and abundant active component, 1, of I. coccinea, thus representing a considerable contribution for promising triterpenoid in cancer chemotherapy. Abbreviations used: MTT: 3,4,5-dimethylthiazol-2-yl-2,5-diphenyltetrazolium bromide, RP: reverse phase, TLC: thin layer chromatography, KOH: potassium hydroxide, IR: infrared, DMF: dimethylformamide, DMSO: dimethyl sulfoxide, TEA: triethylamine, RT: room temperature, EtOAc: ethyl acetate, MeOH: methanol, i-PrOH: iso-propanol, NMR: nuclear magnetic resonance, MDR: multiple drug resistance, RPMI: Roswell Park Memorial Institute
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Comparison of ultrasound-assisted extraction and dynamic maceration over content of tagitinin C obtained from Tithonia diversifolia (Hemsl.) A. gray leaves using factorial design p. 270
Aline M. R. Silva, Nayara L. O. Ferreira, Anselmo E Oliveira, Leonardo L Borges, Edemilson C Conceição
DOI:10.4103/0973-1296.204555  
Background:Tithonia diversifolia belongs to the Asteraceae family. The leaves of T. diversifolia have been studied lately because of the presence of tagitinin C. Objective: Looking for an easy and inexpensive method to extract tagitinin C from T. diversifolia leaves, this work aims to conduct a screening to evaluate the influence of different experimental factors using the dynamic maceration and ultrasound-assisted extraction methods with 23 factorial design based on response surface methodology in enhancing this chemical marker extraction. Materials and Methods: The experimental factors were: extraction time (ET) of 30 and 60 minutes, solid: liquid ratio (SLR) of 5 and 10 grams/grams and ethanolic strength (ES) 48 and 96% (w/w). The experiments were done tripled. The content of tagitinin C in each produced extract was quantified by HPLC method. Results: The highest concentrations of tagitinin C obtained under the experimental design were 0.53 mg/mL and 0.71 mg/mL, respectively for dynamic maceration (DM) and ultrasound-assisted extraction (UAE) from Tithonia diversifolia powdered leaves. For the UAE method, the main parameter for higher contents of tagitinin C was the solid: liquid ratio, followed by the ethanolic strength, and the extraction time was not significant for this method. As for the DM method, all the parameters (SLR, ES, and ET) were significant for a higher content of tagitinin C. Conclusion: Based on the obtained results, it was revealed that the ultrasound-assisted extraction was more effective than dynamic maceration for tagitinin C extraction from T. diversifolia powdered leaves. Abbreviation used: DME: dynamic maceration extraction, UAE: ultrasound-assisted extraction, ET: extraction time, ES: ethanolic strength, SLR: solid:liquid ratio, Tag C: tagitinin C, HPLC: high-performance liquid chromatography.
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Phytochemical and biological evaluations of Arum hygrophilum boiss. (Araceae) p. 275
Fatma U Afifi, Violet Kasabri, Simona Litescu, Ismail F Abaza, Khalid Tawaha
DOI:10.4103/0973-1296.204551  
Background: Arum hygrophilum is a traditional medicinal plant indigenous to Jordan. The present study explores its phytochemistry, antioxidative, antidiabesity, and antiproliferative potentialities. Materials and Methods: Column chromatography and HPLC-MS analysis were used for its phytochemical evaluation. Using leaf crude water and ethanol extracts, the antioxidative capacities, their modulation of pancreatic β-cell proliferation, and insulin secretion as well as glucose diffusion and enzymatic bioassays were evaluated. Results: Three flavonoids (luteolin, isoorientin, and vitexin) and β-sitosterol have been isolated and their structures determined. HPLC-MS analysis of the ethanol extract further revealed the presence of caffeic, ferulic, gallic, and rosmarinic acids and quercetine-3-O-rhamnoside. The ethanol extract exhibited DPPH and ABTS radical scavenging and antioxidative capacities. A. hygrophilum (1), vitexin (2), and rosmarinic acid (3) inhibited pancreatic lipase (PL) dose dependently with PL-IC50 (µg/mL) values in an ascending order: (3); 51.28 ± 7.55 < (2); 260.9 ± 21.1 < (1); 1720 ± 10. Comparable to GLP-1-enhanced β-cell proliferation in 2-day treatment wells, a dose-dependent augmentation of BrdU incorporation was obtained with the A. hygrophilum aqueous extract (AE) (0.5 and 1 mg/mL, with respective 1.33- and 1.41-folds, P < 0.001). A. hygrophilum AE was identified as an inhibitor of α-amylase/α-glucosidase with IC50 value of 30.5 ± 2.1 mg/mL but lacked antiproliferative effects in colorectal cancer cell lines (HT29, HCT116, and SW620) and insulinotropic effects in β-cell line MIN6. Conclusion: A. hygrophilum extracts inhibited gastrointestinal enzymes involved in carbohydrate and lipid digestion and absorption. Abbreviations used:ABTS: 2,2'-Azino-Bis-3-Ethylbenzothiazoline-6-Sulfonic Acid, AE: Aqueous Extract, ANOVA: Analysis Of Variance, AUC: Area Under Curve, BrdU: 5-Bromo-2'-Deoxyuridine, DPPH: 2,2-Diphenyl -1-Pycriylhydrazyl, ELISA: Enzyme Linked Immunosorbent Assay, GLP1: Glucagon Like Peptide 1, GSIS: Glucose Stimulated Insulin Secretion, HPLC-MS: High Performance Liquid Chromatography-Mass Spectrometry, IC50: 50% Inhibitory Concentration, KRH: Krebs/Ringer/Hepes, MTT: 3-(4,5-Dimethylthiazol-2-Yl)-2,5-Diphenyltetrazolium Bromide, OGTT: Oral Glucose Tolerance Test, ORAC: Oxygen Radical Antioxidant Capacity, OSTT: Oral Starch Tolerance Test, PL: Pancreatic Lipase, SEM: Standard Error Of The Mean, SRB: Sulforhodamine B, TEAC: Trolox Equivalent Antioxidant Capacity, TLC: Thin Layer Chromatography
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Effect of Helix aspersa extract on TNFα, NF-κB and some tumor suppressor genes in breast cancer cell line Hs578T p. 281
Ibtissem El Ouar, Cornelia Braicu, Dalila Naimi, Alexendru Irimie, Ioana Berindan-Neagoe
DOI:10.4103/0973-1296.204618  
Background: The garden snail, Helix aspersa, is a big land snail widely found in the Mediterranean countries. It is one of the most consumed species and widely used in zootherapy. Objective: The present study was carried out to investigate for the first time the first time the antitumor activity of an aqueous extract from Helix aspersa. Materials and Methods: The effect of H. aspersa extract was studied on a triple negative breast cancer cell line Hs578T. Firstly, the morphological changes and the mode of cell death induced by the extract have been evaluated by microscopy and acridine orange/ethidium bromide staining. The effect of the extract at dilution 0.1% and 1% was then tested on some genes, regulators of cell death and proliferation like tumor necrosis factor α (TNFα), NF- κB, and the tumor suppressor genes P53 and PTEN. Results: Data demonstrate that the extract induces necrosis in tumor cells. It enhances significantly the expression of TNFα; mRNA levels were 20 and 10 times more important in treated cells compared to nontreated cells. NF-κB and PTEN were inhibited with the dilution 1% after 8 and 24 hours of treatment. P53 expression was further inhibited but only with the highest dose, after 4, 8, and 24 hours. Conclusion: Our results show that H. aspersa extract has an antitumor activity against Hs578T cells; it is a potent stimulator for TNFα and a good inhibitor for NF-κB. Abbreviations used: AO: acridine orange; Bcl-2: B cell lymphoma 2. cDNA: complementary DNA; ELISA: enzyme linked immunosorbent assay; EB: ethidium bromide; IC50: the half maximal inhibitory concentration; mRNA: messenger RNA. MAPK: mitogen-activated protein kinase; NF-κB: nuclearfactorkappa B; PBS: phosphate buffered saline. PI3K: phospho-inositol 3 kinase; PTEN: phosphatase and tensin homolog; ROS: reactive oxygen species. RT-PCR: reverse transcription polymerase chain reaction; TNFα: tumor necrosis factor alpha. TNFR1: TNF receptor-1; TP53: tumor protein 53
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Antimicrobial and anti-inflammatory effects of ethanol extract of Corylopsis coreana uyeki flos p. 286
Da-Eon Park, In-Soo Yoon, Jung-Eun Kim, Ji-Hye Seo, Jin-Cheol Yoo, Chun-Sik Bae, Chang-Dai Lee, Dae-Hun Park, Seung-Sik Cho
DOI:10.4103/0973-1296.204554  
Background: Corylopsis coreana Uyeki (Hamamelidaceae) is a medicinal plant cultivated in Northeast Asia. Previously, we have reported that an ethanol extract of Corylopsis coreana Uyeki flos (ECCF) contains four active compounds with antioxidant activity. Objective: The aim of this study was to investigate the antimicrobial spectrum against infectious bacteria and anti-inflammatory effect of ECCF in a mouse model of acute local inflammation. Materials and Methods: In vitro antimicrobial susceptibility was evaluated using standard plate assay technique. Antimicrobial activities (minimum inhibitory concentration, MIC; μg/mL) were determined with the serial dilution method. In vivo anti-inflammatory activity was studied using a mouse model of carrageenan-induced air pouch inflammation. Results: The ECCF showed antimicrobial activities against general bacteria and drug-resistant bacteria including Staphylococcus aureus, Micrococcus luteus ATCC 9341, Mycrobacterium smegmatis ATCC 9341, Mycrobacterium smegmatis ATCC 9341, Salmonella typhimrium KCTC 1925, and nine methicillin-resistant Staphylococcus aureus strains, with MIC values ranging from 250 to 1000 μg/mL. In in vivo mouse model, inflammatory morphologic changes observed in the air pouch membrane were restored to its normal condition by the ECCF treatment. Moreover, the ECCF significantly reduced exudate volumes, protein contents, inflammatory cell counts, and pro-inflammatory cytokine levels in the exudates recovered from air pouches of the mouse model. Flavonoids in the ECCF were found to contain bergenin, quercitrin, and quercetin with reported anti-inflammatory activity via suppressing tumor necrosis factor-α production. Conclusion: To the best of our knowledge, this is the first report to demonstrate the antimicrobial and anti-inflammatory activities of ECCF. Our results suggest that the ECCF might potentially serve as an alternative or complementary medicine for treating inflammatory diseases caused by microbial infection. Abbreviations used: Corylopsis coreana CCF: Uyeki flos, ECCF: ethanol extract of CCF
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Anti-differentiation effect of B, D-seco limonoids of Swietenia mahogani p. 293
Heejung Yang, Mina Choi, Dong Young Lee, Sang Hyun Sung
DOI:10.4103/0973-1296.204549  
Background: Obesity is a pathological state caused by abnormal or excessive accumulation of fat. Swietenia mahogani JACQ., known as West Indian mahogany, is a medium-sized semi-evergreen tree belonging to Meliaceae. Their seeds are used in Indonesian folk medicine as a treatment for hypertension, diabetes, malaria, and it also has anti-feedant activities. The major components of S. mahogani are B, D-seco limonoids, a type of irregular triterpenes are well known. Objective: We tried to find the bioactive components, which have the inhibitory activity on adipocyte differentiation from the seeds of S. mahogani. Material and Methods: 3T3-L1 cells, derived from mouse preadipocyte, are widely used in studying adipogenesis process. In this study, we used 3T3-L1 cells to find natural products with the inhibitory activity on adipocyte differentiation. S. mahogani seeds were dried and extracted with 100% MeOH. Results: The methanolic extract was fractionated by bioassay-guided method to give nine B, D-seco limonoids (1-9) with slight structural modifications. Among nine compounds, compounds 4, 6 and 8 exhibited significant inhibitory effects of cell differentiation on 3T3-L1 cells. Those compounds have tigloyl residue at C-3 in common. Besides, compounds with no tigloyl residue at C-3 showed insignificant effect. Nevertheless, not all compounds with tigloyl residue at C-3 exerted significant inhibitory effect. Conclusion: These results suggested that tigloyl residue at C-3 may play a role in the anti-proliferative activity on a dipogenesis and the refined extract of S. mahogani may have a potential to be developed as a therapeutic agent to treat obesity.
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Inhibition of cytochrome P450 (CYP3A4) activity by extracts from 57 plants used in traditional chinese medicine (TCM) p. 300
Mohamed L Ashour, Fadia S Youssef, Haidy A Gad, Michael Wink
DOI:10.4103/0973-1296.204561  
Background: Herbal medicine is widely used all over the world for treating various health disorders. It is employed either alone or in combination with synthetic drugs or plants to be more effective. Objective: The assessment of the effect of both water and methanol extracts of 57 widely used plants from Traditional Chinese Medicine (TCM) against the main phase I metabolizing enzyme CYP3A4 in vitro for the first time. Materials and Methods: The inhibition of cytochrome P450 activity was evaluated using a luminescence assay. The principal component analysis (PCA) was used to correlate the inhibitory activity with the main secondary metabolites present in the plant extracts. Molecular modeling studies on CYP3A4 (PDB ID 4NY4) were carried out with 38 major compounds present in the most active plant extracts to validate the observed inhibitory effect. Results: Aqueous extracts of Acacia catechu, Andrographis paniculata, Arctium lappa, Areca catechu, Bupleurum marginatum, Chrysanthemum indicum, Dysosma versipellis, and Spatholobus suberectus inhibited CYP3A4 is more than 85% (at a dose of 100 μg/mL). The corresponding methanol extracts of A. catechu, A. paniculata, A. catechu, Mahonia bealei, and Sanguisorba officinalis inhibited the enzyme by more than 50%. Molecular modeling studies revealed that two polyphenols, namely hesperidin and rutin, revealed the highest fitting scores in the active sites of the CYP3A4 with binding energies equal to -74.09 and -71.34 kcal/mol, respectively. Conclusion: These results provide evidence that many TCM plants can inhibit CYP3A4, which might cause a potential interference with the metabolism of other concomitantly administered herbs or drugs. Abbreviation used: CHARMm: Chemistry at HARvard Macromolecular Mechanics, CYP: Cytochrome P450, DMSO: Dimethyl Sulfoxide, PCA: Principal Component Analysis, PDB: Protein Data Bank, TCM: Traditional Chinese Medicine
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Development and validation of a simultaneous RP-HPLC-UV/DAD method for determination of polyphenols in gels containing S. terebinthifolius raddi (Anacardiaceae) p. 309
Melina G. Carvalho, Cícero F. S Aragão, Fernanda N Raffin, Túlio F. A. de L. Moura
DOI:10.4103/0973-1296.204559  
Topical gels containing extracts of Schinus terebinthifolius have been used to treat bacterial vaginosis. It has been reported that this species has anti-microbial, anti-inflammatory and anti-ulcerogenic properties, which can be attributed to the presence of phenolic compounds. In this work, a sensitive and selective reversed-phase HPLC-UV/DAD method for the simultaneous assay of six polyphenols that could be present in S. terebinthifolius was developed. The method was shown to be accurate and precise. Peak purity and similarity index both exceeded 0.99. Calibration curves were linear over the concentration range studied, with correlation coefficients between 0.9931 and 0.9974. This method was used to determine the polyphenol content of a hydroalcoholic extract and pharmacy-compounded vaginal gel. Although the method is useful to assess the 6 phenolic compounds, some compounds could not be detected in the products. Abbreviations used: RP-HPLC-UV/DAD: Reverse Phase High Performance Liquid Chromatograph with Ultraviolet and Diode Array Detector, HPLC: High Performance Liquid Chromatograph, HPLC-UV: High Performance Liquid Chromatograph with Ultraviolet Detector, ANVISA: Brazilian National Health Surveillance Agency, LOD: Limit of detection, LOQ: Limit of quantitation
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Bioassay-guided isolation and identification of antioxidant flavonoids from Cyclotrichium origanifolium (Labill.) manden and scheng p. 316
Abdussamat Guzel, Huseyin Aksit, Mahfuz Elmastas, Ramazan Erenler
DOI:10.4103/0973-1296.204556  
Background: Medicinal and aromatic plants play a significant role in drug discovery and development process. Flavonoids, revealing a wide spectrum of biological activities, extensively found in plants are important secondary metabolites. Materials and Methods: Aerial parts of Cyclotrichium origanifolium were collected, dried, and boiled in water then extracted with hexane, ethyl acetate, and n-butanol. Total phenolic content, DPPH· scavenging activity, reducing power (FRAP) activity, and ABTS·+ scavenging activity assays were applied for all extracts. The ethyl acetate extract revealing the most antioxidant activity as well as including the highest phenolic contents was subjected to chromatographic techniques (column chromatography, sephadex LH-20, semipreparative HPLC) to isolate the active compounds. The structure of isolated compounds were elucidated by spectroscopic methods (1D NMR, 2D NMR, and LC-TOF/MS). Results: Isosakuranetin (1), eriodictyol (2), luteolin (3), naringenin (4), and apigenin (5) were isolated and identified. All isolated flavonoids displayed the excellent antioxidant activity. Conclusion: The isolated flavonoids and also plant extract have potency to be a natural antioxidant. Abbreviations used: DPPH•: 1,1-diphenyl-2-picryl-hydrazyl free radical, ABTS•+: 2,2•-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid), UV:Ultraviolet, DNA: Deoxyribonucleic acid, BHT: Butylated hydroxytoluene, BHA: Butylated hydroxyanisole, HPLC: High performance liquid chromatography
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Anti-quorum sensing activity of Forsythia suspense on Chromobacterium violaceum and Pseudomonas aeruginosa p. 321
An Zhang, Wei-Hua Chu
DOI:10.4103/0973-1296.204547  
Background: Quorum sensing (QS) plays an important role in the production of virulence factors and pathogenicity in Pseudomonas aeruginosa, and the interruption of QS will be a hopeful pathway to combat bacterial infection. Objective: In this study, we selected Forsythia suspense (Thunb.) Vahl from traditional Chinese herbal medicines for its anti-QS activity. Materials and Methods: Anti-QS of F. suspense extracts (FSE) was monitored using the Chromobacterium violaceum 12472 bioassay. Standard methods were used to investigate the effects of FSE on QS-controlled virulence factors production, swimming motility, and biofilm establishment in P. aeruginosa PAO1. Results: FSE could obviously inhibit the violacein production in C. violaceum 12472 and also could inhibit quorum sensing–regulated virulence factors production and biofilm formation in P. aeruginosa in a concentration-dependent manner. The elastase activity and pyocyanin production were inhibited at a maximum of 40.97 and 47.58% when P. aeruginosa was grown in the presence of 0.25 g/mL FSE, which can also inhibit swimming motility of P. aeruginosa. The biofilm formation ability was decreased about 72.45% when in PAO1 cultured with the 0.25 g/mL FSE. The results suggested that FSE may be used as an alternative drug to control and handle harmful infections caused by bacterial pathogens based on QS inhibition. Abbreviations used: QS: Quorum sensing, Pseudomonas aeruginosa P. aeruginosa, Forsythia suspense F. suspense, FSE: F. suspense extracts, Chromobacterium violaceum 12472 C. violaceum 12472, AIs: autoinducers, AHLs: N-acyl-homoserinelactones, LB: Luria-Bertani, MICs: Minimum inhibitory concentrations, CFU: Colony-Forming Units, ATCC: American Type Culture Collection, PBS: phosphate buffered saline
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Anti-fatigue effect of green tea polyphenols (-)-Epigallocatechin-3-Gallate (EGCG) p. 326
Yu-song Teng, Di Wu
DOI:10.4103/0973-1296.204546  
Background: (-)-Epigallocatechin-3-gallate (EGCG) is the most abundant of the green tea polyphenols that exhibit a variety of bioactivities. The objective of this study was to evaluate the anti-fatigue effect of EGCG by forced swimming exercise. Materials and Methods: The mice were divided into one control group and three EGCG-treated groups. The control group was administered with distilled water and EGCG-treated groups were administered with different dose of EGCG (50, 100, and 200 mg/kg) by oral gavage for 28 days. On the last day of experiment, the forced swimming exercise was performed and corresponding biochemical parameters were measured. Results: The data showed that EGCG prolonged exhaustive swimming time, decreasing the levels of blood lactic acid, serum urea nitrogen, serum creatine kinase and malondialdehyde, which were accompanied by corresponding increase in liver and muscle glycogen contents, and superoxide dismutase, catalase, and glutathione peroxidase activities. Conclusions: This study indicated that EGCG had an anti-fatigue effect. Abbreviations used: EGCG: (-)-Epigallocatechin-3-gallate, ROS: reactive oxygen species, BLA: blood lactic acid, SUN: serum urea nitrogen, SOD: superoxide dismutase, GPx: glutathione peroxidase, CAT: catalase, SCK: serum creatine kinase, MDA: malondialdehyde, C: control, LET: Low-dose EGCG-treated, MET: Middle-dose EGCG-treated, HET: High-dose EGCG-treated, GTE: green tea extract.
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Near-infrared spectroscopy as a process analytical technology tool for monitoring the parching process of traditional chinese medicine based on two kinds of chemical indicators p. 332
Kaiyue Li, Weiying Wang, Yanping Liu, Su Jiang, Guo Huang, Liming Ye
DOI:10.4103/pm.pm_416_16  
Background: The active ingredients and thus pharmacological efficacy of traditional Chinese medicine (TCM) at different degrees of parching process vary greatly. Objective: Near-infrared spectroscopy (NIR) was used to develop a new method for rapid online analysis of TCM parching process, using two kinds of chemical indicators (5-(hydroxymethyl) furfural [5-HMF] content and 420 nm absorbance) as reference values which were obviously observed and changed in most TCM parching process. Materials and Methods: Three representative TCMs, Areca (Areca catechu L.), Malt (Hordeum Vulgare L.), and Hawthorn (Crataegus pinnatifida Bge.), were used in this study. With partial least squares regression, calibration models of NIR were generated based on two kinds of reference values, i.e. 5-HMF contents measured by high-performance liquid chromatography (HPLC) and 420 nm absorbance measured by ultraviolet–visible spectroscopy (UV/Vis), respectively. Results: In the optimized models for 5-HMF, the root mean square errors of prediction (RMSEP) for Areca, Malt, and Hawthorn was 0.0192, 0.0301, and 0.2600 and correlation coefficients (Rcal) were 99.86%, 99.88%, and 99.88%, respectively. Moreover, in the optimized models using 420 nm absorbance as reference values, the RMSEP for Areca, Malt, and Hawthorn was 0.0229, 0.0096, and 0.0409 and Rcalwere 99.69%, 99.81%, and 99.62%, respectively. Conclusions: NIR models with 5-HMF content and 420 nm absorbance as reference values can rapidly and effectively identify three kinds of TCM in different parching processes. This method has great promise to replace current subjective color judgment and time-consuming HPLC or UV/Vis methods and is suitable for rapid online analysis and quality control in TCM industrial manufacturing process. Abbreviations used: NIR: Near-infrared Spectroscopy; TCM: Traditional Chinese medicine; Areca: Areca catechu L.; Hawthorn: Crataegus pinnatifida Bge.; Malt: Hordeum vulgare L.; 5-HMF: 5-(hydroxymethyl) furfural; PLS: Partial least squares; D: Dimension faction; SLS: Straight line subtraction, MSC: Multiplicative scatter correction; VN: Vector normalization; RMSECV: Root mean square errors of cross-validation; RMSEP: Root mean square errors of validation; Rcal: Correlation coefficients; RPD: Residual predictive deviation; PAT: Process analytical technology; FDA: Food and Drug Administration; ICH: International Conference on Harmonization of Technical Requirements for Registration of Pharmaceuticals for Human Use.
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