The Effect And Mechanism Of Low SOCS3 Expression In Promoting STAT3 Pathway Activation And Alleviating Liver Ischemia Reperfusion Injury In Mice

Objective:Liver ischemia-reperfusion injury(LIRI)is not only a common injury during liver transplantation and major liver surgery,but also one of the main factors affecting postoperative outcomes.However,whether it is a major liver disease operation or liver transplantation,the blood flow to the liver will inevitably be blocked during the operation,and the blood supply to the liver will be restored after vascular reconstruction,so liver ischemia-reperfusion injury is an inevitable problem.There is still no reliable way to solve this problem.We identified differentially expressed genes in ischemia-reperfusion injury using dataset GSE12720,and then identified 5 core pathways and 13 hub genes using MCODE,cyto Hubba plug-ins in Cytoscape,and Kyoto Encyclopedia of Genes and Genomes(KEGG).Suppressor of cytokine signaling 3(SOCS3)was selected for subsequent in-depth study.We constructed the AAV vector of SOCS3 by constructing Adeno Associated Virus(AAV)as the vector,and then transfer it into mice to knock it down,and observe the changes of liver inflammation and histopathology after hepatic ischemia-reperfusion.Observe liver inflammation and histopathological changes after hepatic ischemia-reperfusion preliminarily explore of its internal molecular mechanism,to provide new ideas for the clinical improvement of hepatic ischemia-reperfusion injury.Method: In the first part,we screen out hub genes related to liver ischemia reperfusion injury,and SOCS3 was selected as the target gene for subsequent studies to explore the expression of SOCS3 in hepatic ischemia-reperfusion injury.The m RNA and protein expression of SOCS3 were verified in C57 liver ischemiareperfusion mice by q RT-PCR and western blotting.The second part we explore the role and specific molecular mechanism of SOCS3 in liver ischemia-reperfusion in mouse.The injury of liver ischemiareperfusion in mice was detected by serum ALT and AST,and the histopathological conditions of liver ischemia-reperfusion in mice were observed in HE stained sections.We used adeno-associated virus AAV-SOCS3 and its control virus AAV-NC to construct SOCS3 knockdown mice and its control mice,and verified its knockdown effect by q RT-PCR.To explore the role of SOCS3 in liver ischemia-reperfusion injury by establishing a mouse model of liver ischemia-reperfusion injury.In the liver ischemia-reperfusion injury model,we detected serum ALT,AST and TNF-α and IL-6levels in liver tissues,performed HE stained sections to observe liver damage,and detected key signaling molecules of STAT pathway by western blotting,and verified the regulatory mechanism of SOCS3 on liver ischemia-reperfusion injury by observing the protein expression of STAT3.Results:1 we discovered five core pathways and 13 hub genes.2 The expression of SOCS3 was up-regulated in mice with hepatic ischemiareperfusion injury,and the expression was the highest at 6 hours after reperfusion.3 After 6h and 12 h of hepatic ischemia-reperfusion,serum transaminase levels in mice increased significantly.4 After ischemia-reperfusion,the histopathological damage of the liver has appeared at 6 hours after reperfusion,with inflammatory cell infiltration,hepatic sinusoidal congestion,and hepatic lobular structure disorder.5 The recombinant adeno-associated virus successfully infected the liver tissue,and the recombinant gene could be continuously and stably expressed in the liver tissue.6 C57 mice were induced with AAV before surgery.The transcriptional activity and protein of SOCS3 were lower in the knockdown group than in the corresponding empty AAV group,and the difference was statistically significant(P<0.001).7 SOCS3 knockdown mice had significantly lower serum transaminase levels.8 Reduction in histopathological damage to the liver in SOCS3 knockdown mice.9 SOCS3 knockdown mice have reduced levels of inflammatory factors.10 The protein expression of p-STAT3 in AAV-SOCS3 group mice was significantly higher than that of AAV-NC,suggesting that knocking down SOCS3 may inhibit the activation of inflammatory signaling pathways by promoting the expression of phosphorylated STAT3,and relieve Hepatic ischemia-reperfusion injury.Conclusion:1 5 core pathways and 13 hub genes(IL-1β,JUN,TNFAIP3,CCL2,ICAM1,CCL20,SOCS3,IRF1,BIRC3,GADD45 B,MYC,CCL4,and BCL2A1)related to liver ischemia reperfusion injury were successfully obtained.2 Inhibiting the expression of SOCS3 before operation can reduce the liver ischemia-reperfusion injury in mice.3 Low SOCS3 expression may negatively inhibit the activation of IL-6inflammatory signaling pathways by promoting the STAT3 pathway to alleviate hepatic ischemia-reperfusion injury.