Autophagy regulates turnover of lipid droplets via ROS-dependent Rab25 activation in hepatic stellate cell

• Published in: Redox biology 2017-04, Vol.11, p.322-334
• Main Authors: Zhang, Zili, Zhao, Shifeng, Yao, Zhen, Wang, Ling, Shao, Jiangjuan, Chen, Anping, Zhang, Feng, Zheng, Shizhong
• Format: Article
• Language: English
• Subjects: Acetylcysteine - pharmacology; Animals; Antioxidants - pharmacology; Autophagy; Autophagy - drug effects; Autophagy - genetics; Autophagy-Related Protein 5 - agonists; Autophagy-Related Protein 5 - antagonists & inhibitors; Autophagy-Related Protein 5 - genetics; Autophagy-Related Protein 5 - metabolism; Class III Phosphatidylinositol 3-Kinases - genetics; Class III Phosphatidylinositol 3-Kinases - metabolism; Gene Expression Regulation; Glutathione - pharmacology; Hepatic stellate cells; Hepatic Stellate Cells - drug effects; Hepatic Stellate Cells - metabolism; Hepatic Stellate Cells - pathology; Lipid droplets; Lipid Droplets - drug effects; Lipid Droplets - metabolism; Liver - drug effects; Liver - metabolism; Liver - pathology; Liver Cirrhosis - drug therapy; Liver Cirrhosis - genetics; Liver Cirrhosis - metabolism; Liver Cirrhosis - pathology; Male; Mice; Mice, Inbred ICR; Primary Cell Culture; Proteins - antagonists & inhibitors; Proteins - genetics; Proteins - metabolism; Rab25; Reactive Oxygen Species - agonists; Reactive Oxygen Species - antagonists & inhibitors; Reactive Oxygen Species - metabolism; Research Paper; RNA, Small Interfering - genetics; RNA, Small Interfering - metabolism; ROS; Signal Transduction
• ISSN: 2213-2317; 2213-2317
• DOI: 10.1016/j.redox.2016.12.021

Activation of hepatic stellate cells (HSCs) is a pivotal event in liver fibrosis, characterized by dramatic disappearance of lipid droplets (LDs). Although LD disappearance has long been considered one of the hallmarks of HSC activation, the underlying molecular mechanisms are largely unknown. In this study, we sought to investigate the role of autophagy in the process of LD disappearance, and to further examine the underlying mechanisms in this molecular context. We found that LD disappearance during HSC activation was associated with a coordinate increase in autophagy. Inhibition or depletion of autophagy by Atg5 siRNA impaired LD disappearance of quiescent HSCs, and also restored lipocyte phenotype of activated HSCs. In contrast, induction of autophagy by Atg5 plasmid accelerated LD loss of quiescent HSCs. Importantly, our study also identified a crucial role for reactive oxygen species (ROS) in the facilitation of autophagy activation. Antioxidants, such as glutathione and N-acetyl cysteine, significantly abrogated ROS production, and in turn, prevented autophagosome generation and autophagic flux during HSC activation. Besides, we found that HSC activation triggered Rab25 overexpression, and promoted the combination of Rab25 and PI3KCIII, which direct autophagy to recognize, wrap and degrade LDs. Down-regulation of Rab25 activity, using Rab25 siRNA, blocked the target recognition of autophagy on LDs, and inhibited LD disappearance of quiescent HSCs. Moreover, the scavenging of excessive ROS could disrupt the interaction between autophagy and Rab25, and increase intracellular lipid content. Overall, these results provide novel implications to reveal the molecular mechanism of LD disappearance during HSC activation, and also identify ROS-Rab25-dependent autophagy as a potential target for the treatment of liver fibrosis. [Display omitted] •Autophagosome generation and autophagic flux are increased during HSC activation.•The inhibition of autophagy blocks LD disappearance of quiescent HSCs.•The induction of autophagy accelerates LD disappearance of quiescent HSCs.•Rab25 activation is required for autophagy to degrade LDs during HSC activation.•Mitochondrial H2O2 production triggers autophagy activation during HSC activation.