Knockdown Of Rap-1 Regulates Non-alcoholic Steatohepatitis Via Sirt1/AMPK-mediated Autophagy Induction

Objective: To explore the expression of Rap-1 in Non-alcoholic steatohepatitis(NASH),and to clarify the biological roles and molecular mechanism of Rap-1 in NASH.Rap-1 may serve as a novel therapeutic target for NASH.Methods: The expression of Rap-1 in liver tissue from NASH patients and normal was evaluated by Western Blot,RT-PCR and immunohistochemistry staining.And we investigated the association between the degree of Rap-1 and the severity of NASH.Then,wild-type(WT)mice and Rap-2 knockout(KO)mice were randomly divided into the normal diet group and HFD/MCD group to build animal models.At the animal level,liver /body weight ratio,carbohydrate and lipid metabolism change and lipid deposition,fibrosis and inflammation level in liver were measured.ELISA,Western Blot,RT-PCR,immunohistochemistry staining,immunofluorescence staining,Oil red O staining was used to explore the biological functions of Rap-1 in NASH progression.We filtered related genes Sirt1 and AMPK in NASH by High throughput and RNA-seq,and further verified by Western Blot,Ch-IP.Results: Rap-1 expression is significantly diminished in NASH patient liver samples and in mice with NASH.Rap-1 may alleviate NASH,and knockdown the Rap-1 can aggravate the degree of NASH in mice induced by HFD/MCD.Rap-1 deficiency can influence the development of NASH by regulating autophagy in hepatocytes.Conclusion: Rap-1 can regulate the level of lipid metabolism,inflammation and fibrosis in hepatocytes by autophagy,inhibiting the progression of NASH.We propose Rap-1 may as a potential therapeutic target for NASH.