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Journal of Clinical and Experimental Hepatology logoLink to Journal of Clinical and Experimental Hepatology
. 2011 Nov 9;1(2):57–67. doi: 10.1016/S0973-6883(11)60123-0

Phyllanthin of Standardized Phyllanthus amarus Extract Attenuates Liver Oxidative Stress in Mice and Exerts Cytoprotective Activity on Human Hepatoma Cell Line

Rajesh Krithika *, Ramtej J Verma *,**,*, Pranav S Shrivastav †, Lonchin Suguna *
PMCID: PMC3940533  PMID: 25755316

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.

Aim

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. Conclusion: The study established a significant liver-protecting role of standardized P. amarus extract due to the presence of active ingredient phyllanthin.

Keywords: Hepatitis B virus, hepatoprotection, HepG2, high performance liquid chromatography, high performance thin layer chromatography, lipid peroxidation

Abbreviations: 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; 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

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