EFFECT OF UBIQUITIN-SPECIFIC PROTEASE 13(USP13)KNOCKOUT ON ACETAMINOPHEN-INDUCED HEPATIC INFLAMMATORY STRESS IN MICE
Objective To investigate the role of ubiquitin-specific protease 13(USP13)in hepatic inflammatory stress response induced by acetaminophen(APAP).Methods Usp13+/+mice and Usp13-/-mice were set up as the control group and three experimental groups(APAP 2 h,4 h and 12 h group),with 6 mice in each group.After being starved for 18 h,the liver inflammatory stress model was established by intraperitoneal injection of 300 mg/kg APAP in the experimental groups.Live tissues and serum were collected at 2,4,and 12 h respectively.In the control group liver tissues and serum were collected at 0 h.Histopathological changes in the liver were observed by HE staining.Inflammatory cell infiltration in liver tissue was observed by immunohistochemistry.The levels of alanine aminotransferase(ALT)and aspartate aminotransferase(AST)in the serum were measured by ELISA.Glutathione(GSH)kit was used to detect hepatic GSH changes.Western blot was used to detect hepatic cytochrome p450 2E1(CYP2E1)protein expression and c-Jun N-terminal kinase(JNK)pathway activation.The expression of inflammatory factor-related genes was detected by qPCR.Results Compared with the control group,mice in the APAP 2 h and APAP 4 h groups showed no significant changes in hepatic CYP2E1 protein expression,a significant decrease in GSH(P<0.0001),an increase in JNK pathway activation and inflammatory factor expression(P<0.001).Compared with the control group,mice in the APAP 12 h group experienced significant liver necrosis,increased number of F4/80+ cells(P<0.001)and Ly6G+ cells(P<0.01),as well as increased serum ALT and AST levels(P<0.01).However,Usp13 knockout did not significantly affect the pathophysiological process of APAP-induced inflammatory stress response in the liver compared with Usp13+/+ mice in all experimental groups.Conclusion USP13 deletion does not affect various pathophysiological processes during the APAP-induced inflammatory stress response in the liver,which differs from previous reports showing the involvement of USP13 in the development of multiple liver diseases and suggests the complexity of the role of USP13 in liver diseases.[ACTA NUTRIMENTA SINICA,2024,46(1):56-61]