Glutathione ameliorates Hypoxia/Reoxygenation (H/R) induced hepatocyte oxidative damage via regulating HO-1 signaling

Wuye Chen,Xiaozai Luo,Shaomei Zhu,Yongping Fang,Zhengyu Liu,Zhengyuan Xia,Kunping Li,Yihong Wang

doi:10.1590/fst.61221

Wuye Chen, Xiaozai Luo + Show 6 more

Open Access

https://doi.org/10.1590/fst.61221

Copy DOI

Abstract

In our study, CCK-8 assay and LDH release detection were performed to detect the optimal protective concentration of GSH on HL7702 cell viability during H/R injury. HL7702 cells were randomly divided into four groups: Control group, H/R group, H/R+GSH group, and H/R+GSH+HO-1-siRNA group. Then, reactive oxygen species (ROS) was evaluated by DHE staning, MDA, T-SOD measurements; Cell injury was detected by CCK-8, LDH release, and supernatant AST and ALT levels; Apoptosis was determined by Hoechst staining and caspase 3 level. Compared with controls, H/R caused significant HL7702 cell injury evidenced as reduced cell viability, increased LDH release and apoptotic cell death (P < 0.01), with concomitant increases in ROS and MDA production (P < 0.01), while treated with GSH in H/R group significantly attenuated oxidative injury, enhanced cell viability and downregulated cell apoptosis (P < 0.01) together with HO-1 upregulation (P < 0.01). Knockdown HO-1 by its siRNA cancelled the protective effects of GSH from H/R compared with GSH group (P < 0.01). HO-1 was induced in HL7702 exposed to H/R injury and its level was obviously overexpressed after H/R injury with GSH treatment, suggesting its protective potential in GSH against H/R injury. GSH increases the expression of HO-1, which enhanced the early antioxidative activity and played a protective role against HL7702 cells H/R injury.

Highlights

Ischemia/reperfusion (I/R) injury may occur in various organs, which is a common clinical problem
The molecular mechanisms mediating hepatocellular hypoxia/ reoxygenation (H/R) injury are largely unknown, activation of cell apoptosis play an important role in its pathology (Ge et al, 2019; Katwal et al, 2018)
The results of western blot assay showed that the expression of Heme Oxygenase-1 (HO-1) in H/R hepatocytes could be regulated by GSH or HO-1 siRNA

Summary

Introduction

Ischemia/reperfusion (I/R) injury may occur in various organs, which is a common clinical problem. Liver is sensitive to ischemia and hypoxia, how to effectively relieve postoperative hepatic ischemia-reperfusion injury (HIRI) has become a hot topic in clinical research (Pagano et al, 2018; Rakić et al, 2018). The molecular mechanisms mediating hepatocellular H/R injury are largely unknown, activation of cell apoptosis play an important role in its pathology (Ge et al, 2019; Katwal et al, 2018). A process of programmed cell death, significantly contributes to both hepatocellular loss during liver I/R injury and liver failure after ischemia (Gao et al, 2018). Animal studies have demonstrated that antioxidants can improve HIRI injury by increasing expression of antioxidant enzymes, and increasing antioxidant capacity during early phase of I/R may be a potential therapeutic approach to reduce the incidence of HIRI and hepatic fibrosis

Objectives

Methods

Results

Conclusion