In vitro assessment of hepatoprotective agents against damage induced by acetaminophen and CCl4

Liliana Torres González,Ricardo Salazar Aranda,Jonathan Pérez Meseguer,Linda Elsa Muñoz Espinosa,Paula Cordero Pérez,Noemí Waksman Minsky

doi:10.1186/s12906-016-1506-1

Abstract

BackgroundIn vitro bioassays are important in the evaluation of plants with possible hepatoprotective effects. The aims of this study were to evaluate the pretreatment of HepG2 cells with hepatoprotective agents against the damage induced by carbon tetrachloride (CCl4) and paracetamol (APAP).MethodsAntioxidative activity was measured using an assay to measure 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging. The in vitro hepatotoxicity of CCl4 and APAP, and the cytotoxic and hepatoprotective properties of silymarin (SLM), silybinin (SLB), and silyphos (SLP) were evaluated by measuring cell viability; activities of aspartate aminotransferase (AST), alanine aminotransferase (ALT), and lactate dehydrogenase (LDH); total antioxidant capacity (TAOxC); and reduced glutathione (GSH), superoxide dismutase (SOD), and lipid peroxidation (malondialdehyde (MDA) levels).ResultsOnly SLB and SLM showed strong antioxidative activity in the DPPH assay (39.71 ± 0.85 μg/mL and 14.14 ± 0.65 μg/mL, respectively). CCl4 induced time- and concentration-dependent changes. CCl4 had significant effects on cell viability, enzyme activities, lipid peroxidation, TAOxC, and SOD and GSH levels. These differences remained significant up to an exposure time of 3 h. APAP induced a variety of dose- and time-dependent responses up to 72 h of exposure. SLM, SLB, and SLP were not cytotoxic. Only SLB at a concentration of 100 μg/mL or 150 μg/mL significantly decreased the enzyme activities and MDA level, and prevented depletion of total antioxidants compared with CCl4.ConclusionsCCl4 was more consistent than APAP in inducing cell injury. Only SLB provided hepatoprotection. AST, LDH, and MDA levels were good markers of liver damage.

Highlights

In vitro bioassays are important in the evaluation of plants with possible hepatoprotective effects
The aims of this study were to evaluate the hepatoprotective activities of SLM, SLB, and SLP against liver damage induced by APAP and Carbon tetrachloride (CCl4) in the Human hepatoma cell line (HepG2) cell line
In this study, we used the HepG2 cell line to evaluate the hepatoprotective activity of SLM, SLB, and SLP against liver damage induced by APAP and CCl4 in an attempt to establish a simple strategy for monitoring the hepatoprotective activities of plant extracts without high-end and excessive testing

Summary

Introduction

In vitro bioassays are important in the evaluation of plants with possible hepatoprotective effects. Medicinal plants with hepatoprotective activity contain a large number of bioactive molecules. One hepatoprotective agent used widely in the treatment of various liver disorders, such as hepatitis or fatty infiltration caused by alcohol or toxins, is the standardized extract of Silybum marianum, known as milk thistle or silymarin (SLM) [8,9,10,11]. It is a complex mixture of the flavonolignans silybinin (SLB), silychristin, silydianin, and isosilybin. SLM is poorly absorbed, the bioavailability of SLB is higher than that of phosphatidylcholine (silyphos (SLP)) [14, 15]

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