Optimized models of xenobiotic-induced oxidative stress in HepG2 cells

Yollada Sriset,Kanokwan Jarukamjorn,Waranya Chatuphonprasert

doi:10.4314/tjpr.v18i5.13

Abstract

Purpose: To evaluate the molecular impact of ethanol, sodium selenite, and tert-butyl hydroperoxide (TBHP) on oxidant-antioxidant balance in HepG2 cells to establish an optimized oxidative stress model of HepG2 cells. Methods: HepG2 cells were treated with ethanol (10 - 500 mM) and sodium selenite (1 - 10 µM) for 24 and 48 h and with TBHP (50 - 200 µM) for 3 and 24 h, respectively. Biomarkers for cellular injury, ie, lactate dehydrogenase (LDH), aspartate aminotransferase (AST), alanine aminotransferase (ALT), and malondialdehyde (MDA), and for antioxidant system, i.e., superoxide dismutase (SOD), catalase (CAT), and total glutathione content, were determined. Results: All treatments increased the levels of LDH, AST, ALT, and MDA but decreased SOD and CAT activities and the total glutathione content in HepG2 cells. Oxidative stress was induced by these oxidative stressors in HepG2 cells via oxidant-antioxidant imbalance, with TBHP (100 µM, 3 h) acting as a powerful oxidant based on the minimal time to induce oxidative stress. The antioxidants, ascorbic acid and gallic acid, improved oxidant-antioxidant imbalance against xenobiotic-induced oxidative stress in HepG2 cells. Conclusion: These oxidative stress models are suitable for investigating the antioxidant and/or hepatoprotective potential of chemicals, including natural compounds.

Highlights

The liver plays a key role in detoxification, including the regulation of physiological homeostasis damage to this organ results in bodily dysfunction [1]
-©---2-0--1--9---T--h-e---a--u-t-h--o-r-s--.--T-h--i-s--w--o--r-k--i-s--l-i-c-e--n-s--e--d--u--n--d-e--r--t-h-e---C--r-e--a-t-i-v-e---C--o--m--m---o-Tn--rs-o-A-p-t-tJ-r-iP-b-uh--ta-io-r-nm---4R-.-0e--s-I,n--Mt-e--ra-ny-a--2t-i0o--1n--9a-;l--L1--i8c--(e-5n-)-s:--e1--0--0-1 of such xenobiotics include ethanol and sodium selenite, which may be introduced through the diet and lifestyle, and tert-butyl hydroperoxide (TBHP), which is typically related to industrial air pollution [4]
All concentrations of sodium selenite significantly increased the levels of these biomarker enzymes at both 24 and 48 h (Figure 1 E and H)

Summary

Introduction

The liver plays a key role in detoxification, including the regulation of physiological homeostasis damage to this organ results in bodily dysfunction [1]. Toxic compounds are among the most pivotal causes of liver toxicity due to the generation of free radical byproducts, especially reactive oxygen species (ROS) [2]. -©---2-0--1--9---T--h-e---a--u-t-h--o-r-s--.--T-h--i-s--w--o--r-k--i-s--l-i-c-e--n-s--e--d--u--n--d-e--r--t-h-e---C--r-e--a-t-i-v-e---C--o--m--m---o-Tn--rs-o-A-p-t-tJ-r-iP-b-uh--ta-io-r-nm---4R-.-0e--s-I,n--Mt-e--ra-ny-a--2t-i0o--1n--9a-;l--L1--i8c--(e-5n-)-s:--e1--0--0-1 of such xenobiotics include ethanol and sodium selenite, which may be introduced through the diet and lifestyle, and tert-butyl hydroperoxide (TBHP), which is typically related to industrial air pollution [4]. These xenobiotics have been used as oxidative stress stimuli, causing hepatotoxicity [5,6,7] via the induction of cellular oxidative stress [8,9], neither the concentrations nor the trial intervals were optimized in these previous studies

Results

Discussion

Conclusion