title: Phyllanthin of Standardized Phyllanthus amarus Extract Attenuates Liver Oxidative Stress in Mice and Exerts Cytoprotective Activity on Human Hepatoma Cell Line.
abstract: BACKGROUND
  Phyllanthus amarus, a traditional herbal liver-protecting medicine, is known to contain an active ingredient phyllanthin. Many research studies and clinical trials performed in the past using this plant have given contentious results which clearly accentuates the need for the standardization of the extracts.
  
  
  OBJECTIVE
  In this study, P. amarus extract was standardized for phyllanthin content by high performance thin layer chromatography (HPTLC) and high performance liquid chromatography (HPLC) analysis. The preventive role of a standardized extract of P. amarus against CC14-induced hepatotoxicity in vivo and in vitro using mice model and human hepatoma HepG2 cell line, respectively, was investigated.
  
  
  METHODS
  Phyllanthin was used as a marker phytochemical for the standardization of P. amarus extract. The extracts were verified for phyllanthin content by HPTLC and HPLC. Female mice were orally administered with CCl4 either with or without standardized P. amarus extract in three different doses. Similarly, the cytoprotective role of the standardized extract in vitro was studied in HepG2 cell line.
  
  
  RESULTS
  Oral administration of CCl4 resulted in increased oxidative stress, decreased antioxidative defense, and liver injury. Treatment with P. amarus along with CCl4 significantly mitigated the increase in activities of liver marker enzymes, lipid peroxidation, and bilirubin content. It also increased the antioxidant enzymatic and non-enzymatic defense parameter levels. The results of the in vitro study conducted in HepG2 cells indicated that the hepatotoxin lowered 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (Mil) metabolism and increased the release of transaminases which were corrected with co-incubation with P. amarus.
  
  
  CONCLUSIONS
  The study established a significant liver-protecting role of standardized P. amarus extract due to the presence of active ingredient phyllanthin.
authors.affiliation: Department of Biochemistry, Central Leather Research Institute, Council of Scientific and Industrial Research (CSIR), Chennai, India.
authors.forename: Rajesh
authors.initials: R
authors.lastname: Krithika
authors.affiliation: Department of Biochemistry, Central Leather Research Institute, Council of Scientific and Industrial Research (CSIR), Chennai, India ; Zoology Department, Gujarat University, Ahmedabad, India.
authors.forename: Ramtej J
authors.initials: RJ
authors.lastname: Verma
authors.affiliation: Chemistry Department, School of Sciences, Gujarat University, Ahmedabad, India.
authors.forename: Pranav S
authors.initials: PS
authors.lastname: Shrivastav
authors.affiliation: Department of Biochemistry, Central Leather Research Institute, Council of Scientific and Industrial Research (CSIR), Chennai, India.
authors.forename: Lonchin
authors.initials: L
authors.lastname: Suguna
keywords: ALT, alanine transaminase; ANOVA, analysis of variance = AST = aspartate transaminase; CAM, complementary and alternative medicines; CAT, catalase; DMSO, dimethylsulfoxide; GSH, glutathione; HBV, hepatitis B virus; HETP, height equivalent of theoretical plates; HPLC, high performance liquid chromatography; HPTLC, high performance thin layer chromatography; HQC, high quality control; HepG2; Hepatitis B virus; LDH, lactate dehydrogenase; LPO, lipid peroxidation; LQC, low quality control; MDA, malondialdehyde; MTT, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide; TAA, total ascorbic acid; hepatoprotection; high performance liquid chromatography; high performance thin layer chromatography; lipid peroxidation