The Effect And Mechanism Of The MicroRNA-122 And SOCS3 In Sepsis Induced Liver Cell Injury

Objective:To investigate the expression and role of miR-122 in sepsis induced liver cell injury,and the related mechanisms of SOCS3 and NF-κB.Methods1.LO2 cells were divided into two groups:LO2 group(normal untreated)and LO2+LPS group(LPS induced group,10-3g/L);the expression of miR-122,socs3 m RNA and NF-κB m RNA was detected by RT-q PCR.2.The LO2 cells were transfected with miR-122 mimics or miR-122 inhibitor separately.To confirm whether there were overexpression and inhibition effect,RT-q PCR was used to assay the expressions of miR-122 in cells.3.The LO2 cells were transfected with miR-122 mimics or miR-122 inhibitor separately.LPS(10-3g/l)was added after 24 hours.MTT was used to detect the proliferation of cells,Annexinv/PI double staining was used to detect the apoptosis of cells,RT-q PCR was used to assay the expressions of SOCS3 m RNA and NF-κB m RNA in cells,Western Blot was used to detect the protein expressions of SOCS3 and NF-κB.Results:1.The expression of miR-122,SOCS3 m RNA and NF-κB m RNA in LO2 cells induced by LPS.Compared with normal LO2 group,the expression of miR-122 and SOCS3 m RNA was down regulated and the expression of NF-κB was up-regulated in LPS induced LO2+LPS group.2.After transfection of miR-122 mimics and miR-122 inhibitor into LO2 cells,the expression of miR-122 was detected by RT-q PCR.Compared with the control group(LO2 mimics NC group),the expression of miR-122 in the experimental group(LO2 mimics group)was increased,and compared with the control group(LO2 inhibitor NC group),the expression of miR-122 in the experimental group(LO2 inhibitor group)was decreased,which indicated that overexpression and interference were effective,and further experiments could be carried out.3.Transfection of miR-122 mimics into LO2 cells,after LPS induced injury,cell proliferation,apoptosis,SOCS3,NF-κB expression:compared with the normal control group(mimics NC group),the growth rate of cells in mimics transfection group was slightly faster,the apoptosis rate was reduced,SOCS3 expression was up-regulated,NF-κB expression was down regulated;compared with the negative control group(mimics NC+LPS group),the growth rate of cells in mimics+LPS group was lower.There was no significant difference in the expression of NF-κB and SOCS3.4.Transfection of miR-122 inhibitor into LO2 cells,after LPS induced injury,cell proliferation,apoptosis,SOCS3 expression,NF-κB expression.Compared with the negative control group(inhibitor NC group),the growth of cells in the inhibitor transfection group was slower,the number of apoptosis increased,SOCS3 expression was down regulated,NF-κB expression was up-regulated;compared with the normal control group(inhibitor NC+LPS group),the growth of cells in the inhibitor+LPS group was slower.There was no significant difference in the expression of SOCS3 and NF-κB between the two groups.Conclusion:MiR-122 may further inhibit the activation of NF-κB by regulating the expression of SOCS3,which provides a new target for the diagnosis and treatment of liver injury in sepsis.