Mechanisms And Therapeutic Treatment Of Endoplasmic Reticulum-Associated Deficiency Induced Liver Injury

Objective:Liver injury is the most common type of liver disease and is considered a predisposition to many liver disease.The causes of liver injury have been deeply explored.However,the molecular mechanisms triggering liver injury are still unclear.Endoplasmic Reticulum-associated deficiency(ERAD)is a protein quality control system in eukaryotic cells responsible for clearing unfolded proteins and restoring cell homeostasis.It is unclear whether ERAD plays any role in liver injury.Therefore,the objective of this study is to investigate the role of ERAD dysfunction during the pathogenesis of liver injury and to explore the potential role of Mg2+in the prevention of liver disease.Methods:1.Use CRISPR-Cas9 technology to construct HepG2 cells with ERAD deficiency.2.Liver injury was detected by analyzing plasma biochemical indexes and tissue sections of normal mice and ERAD deficient mice.3.Western blot and Co-IP were used to analyze the degradation and aggregation of unfolded proteins in ERAD-deficient HepG2 cells.4.Fat metabolism in HepG2 cells with ERAD deficiency was analyzed by oil red O staining.5.Mg2+ intervention was used to detect the changes in relative cell activity of ERAD-deficient HepG2 cells.Results:1.HepG2 cells and mouse models with ERAD deficiency were successfully constructed.2.Loss of ERAD function led to liver injury in mice.3.Impaired degradation of unfolded proteins in ERAD-deficient HepG2 cells.4.Aggregation of unfolded proteins occurred in ERAD-deficient HepG2 cells.5.Unfolded proteins are substrates of ERAD and are degraded by the ubiquitin pathway.6.Abnormal fat metabolism occurred in ERAD-deficient HepG2 cells.7.Mg2+ intervention can restore the relative decline in cell activity caused by ERAD dysfunction.Conclusions:The present findings suggest that ERAD plays an important role in supporting hepatocellular functions through maintaining endoplasmic reticulum homeostasis in hepatocytes.The present study also provide evidence that Mg2+supplementation may be used as a potential approach for the prevention and treatment of liver injury.