The Study Of The Expression Of Inflammatory Cytokines And Changes Of Regenerative Cell Cycle In Liver Injury

Objective: To research the changes of inflammatory cytokines IL-6,MCP-1,TGF-?,TNF-? in liver injury,and regenerative cell cycle after hepatocytes injured,the study aims to explore the relationship betweeninflammatory factors and cell regeneration in liver injury,to provide ideas for the clinical treatment of liver diseases.Methods 1.Hep G2 cells were cultured in DMEM medium.When the cells were grown to the logarithmic phase,treated them with different concentrations of ethanol.They were divided into three groups(blank control group,0.001 M,0.01M).The expression of the targetm RNA in cells was detected by q RT-PCR.2.Hep3 B cells were cultured in MEM medium and Hep G2 and L02 cells were cultured in DMEM medium.Threetypes of cells were treated with different concentrations of ethanol or carbon tetrachloride for 24 hours,and the cell cycle changes were measured by flow cytometry.L02 cells were treated with different concentrations of carbon tetrachloride for 3 hours,6 hours,12 hours and 24 hours respectively.The cell cycle changes were measured.Used SPSS 17.0 statistical software for data analysising and processing,each sample at least three experiments.Results:1 Real-Time PCR results Contrasted the blank control group,the expression level of IL-6 m RNA was increased at the concentration of 0.001 M and 0.01 M,and the difference between 0.01 M group and control group was statistically significant(with actin as control,P<0.05).Contrasted the blank control group,the expression level of MCP-1 m RNA was increased at the concentration of 0.001 M and 0.01 M,and the difference between the 0.01 M group and the control group was statistically significant(with actin as the internal reference,P <0.01).Contrasted the blank control group,the expression of TGF-? m RNA in 0.001 M group and 0.01 M group increased,and the difference was significant at the concentration of 0.001 M.,P <0.001.Contrasted the blank control group,the expression level of TNF-? m RNA in 0.001 M group and 0.01 M group increased,and the difference was significant at the concentration of0.001 M,P<0.05.2 Cell cycle test results Used different concentrations of ethanol or carbon tetrachloride treated Hep3 B,Hep G2 and L02 cells,then the proportion of G2 phase cells in each group was increased.After 3,6,12,24 hours treatment of different concentrations of CCl4,the percentage of L02 cells in G2 phase was significantly increased.Compared with the untreated,the higher the concentration of CCl4 and the longer the treatment timewas,the more obviously the proportion of G2 phase cells was,P <0.05,the difference was statistically significant.Conclusion: The expression of IL-6,MCP-1,TGF-? and TNF-? in liver injury was significantly increased after liver injury.The proportion of G2 / M phase cells in the cells after injury increased.Suggesting that these inflammatory mediators and cytokines can activate and regulate the signal pathway in the process of liver regeneration,and further study should be carried out on the specific mechanism.