Resarch Of IL-6 In Liver Regeneration After Apap-induced Acute Liver Injury

Objective: To explore the role of IL-6 in the process of liver regeneration after APAP-induced acute liver injury in vitro experiments.Methods: Using in vitro experiments,AML12 cell lines which origins from normal mice hepatocytes,cultured and treated with different concentrations of APAP drugs and IL-6 neutralizing antibody（IL-6 Ab）.CCK-8 method was used to detect cell viability and screen out the appropriate drug effects concentration.The IL-6 concentration was measured by ELISA.Western blot was used to detect the expression levels of liver regeneration-related proteins such as PCNA,Cyclin D1 and HGF;the m RNA levels of IL-6,PCNA and Cyclin D1 were measured by q RT-PCR.Datas were analyzed by Graph Pad prism8.0 software.Results:1.Based on the CCK-8 data,the optimal drug concentration of 5mmol/L APAP for modeling and 0.01 μg/ml IL-6 Ab acting cells were selected.2.The results of ELISA showed that the IL-6 concentrations at 0,4,12,24,and 48 h were 0,1.794,2.264,1.658,and 1.086 pg/ml,respectively..3.Western blot results showed that compared with other time points after APAP administration on AML12 cells,the protein levels of Cyclin D1 was the highest at 4 h,PCNA and HGF were the highest at 12 h（P<0.05）.As compared with the 4 h APAP group,the expression of Cyclin D1 and p-STAT3 protein in the APAP+IL-6 Ab group decreased（P<0.05）;compared with the 12 h APAP group,the expression of PCNA and HGF in the APAP+IL-6 Ab group decreased（P<0.05）.4.The results of q RT-PCR showed that the m RNA levels of IL-6,PCNA,and Cyclin D1 in the 4 h APAP group reached the peak（P<0.05）;compared with the 4 h APAP group,the PCNA and Cyclin D1 levels in the APAP+IL-6 Ab group were reduced（P<0.05）.Conclusions: IL-6 Ab inhibits STAT3 phosphorylation by neutralizing IL-6 and subsequently inhibits liver regeneration.It is speculated that IL-6 may play a role in promoting liver regeneration after APAP-induced liver injury.