Anti-apoptotic effects of phyllanthin against alcoholinduced liver cell death

Wanida Sukketsiri,Kitja Sawangjaroen,Supita Tanasawet

doi:10.4314/tjpr.v15i5.12

Abstract

Purpose: To evaluate the anti-apoptotic effect of phyllanthin on alcohol-induced liver cell death in HepG2 cells alone and in co-culture with human monocytic (THP-1) differentiated macrophage cells.Methods: Cell viability was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The cells were pretreated with 1, 5 and 10 μM phyllanthin for 24 h followed by 1300 mM alcohol for HepG2 cells and 2000 mM alcohol for the co-cultured cells. Thereafter, intracellular reactive oxygen species (ROS), mitochondrial membrane potential (MMP) changes, apoptotic cell death and caspase-3/7 activities were assessed.Results: Alcohol exposure significantly increased intracellular ROS generation (p < 0.001), decreased MMP changes (p < 0.001), increased the number of apoptotic and necrotic cells (p < 0.001) and also induced higher caspase-3/7 activity (p < 0.001) in the co-culture with THP-1 differentiated macrophage cells than in HepG2 cells alone. Pretreatment of HepG2 cells and co-cultured cells with phyllanthin for 24 h prior to alcohol exposure significantly decreased intracellular production of ROS (p < 0.001) and also increased the change in MMP (p < 0.001) as well as caused a decrease in the number of apoptotic and necrotic cells (p < 0.001), but inhibited caspase-3/7 activity (p < 0.001).Conclusion: The results indicate that phyllanthin treatment may have a significant therapeutic effect on alcohol-related liver diseases.Keywords: Hepatoprotective, Liver diseases, Human monocytic cells, Reactive oxygen species, Apoptosis, Co-culture, Mitochondrial membrane potential

Highlights

Alcohol consumption is a major cause of chronic liver disease which may develop after years of chronic alcohol consumption and results in high morbidity and mortality rates [1]
Alcohol produced a significant increase in the intracellular reactive oxygen species (ROS) production to 122.66 ± 2.56 and 126.43 ± 4.52 % of the control in the HepG2 and co-cultured cells, respectively after 4 h of alcohol exposure (Figure 2)
In the hepatocytes cultured alone, these results indicated that alcohol caused an elevation in the intracellular ROS level; it was significantly reduced by phyllanthin pretreatment

Summary

Introduction

Alcohol consumption is a major cause of chronic liver disease which may develop after years of chronic alcohol consumption and results in high morbidity and mortality rates [1]. Alcohol-induced liver injury is a complex process and is dependent upon the interplay of multiple cell types in the liver. It is of interest, that hepatic macrophages (Kupffer cells) play a pivotal role in the pathogenesis of alcoholinduced liver cell death [3]. Current alcohol liver disease therapy using modern medical practices is often restricted in its efficacy, has high adverse events and is costly [1]. The mechanism of the hepatoprotective activity of phyllanthin has been shown to effectively facilitate the recovery of its antioxidant capability including the level of total glutathione, the activities of superoxide dismutase and glutathione reductase [12,13]. The effect of phyllanthin on alcohol-induced cell death has not been clarified

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