Exploration of molecular mechanism of Jianpi Yanggan Decoction in treating autoimmune hepatitis based on Th17/Treg axis
Objective To explore the mechanism of action of Jianpi Yanggan Decoction in treating autoimmune hepatitis based on helper T cell 17(Th17)/regulatory T cells(Treg)axis.Methods Balb/c mice were randomly divided into 4 groups,namely the control group,the model group,Prednisolone group,and Jianpi Yanggan Decoction group.The modeling method was performed by injecting Concanavalin A(ConA)into the tail vein once a week for 6 consecutive weeks.At the same time,corresponding drugs were given by gavage once a day for 6 consecutive weeks.The differences in the ultrastructure of liver tissues among different groups under electron microscopy scanning were observed.Flow cytometry was used to detect the ratios of Th17 and Treg cell subsets in the spleen and peripheral blood.Flow fluorescence assay was used to detect the levels of inflammatory cytokines[interleukin(IL)-2,IL-6,IL-17A,tumor necrosis factor-α(TNF-α),IL-4,IL-10]in plasma.And immunohistochemistry was used to detect the expression of transcription factors RORγt and Foxp3 in liver tissue.Results Electron microscopy observation:In the control group,hepatocyte cords and the structue of hepatic sinusoids were clear,and surface of liver sinuysoidal endothelial cells was smooth,with a uniform distribution of suprafenestral networks.In the model group,hepatocyte cords,the structure of hepatic sinusoids,and fenestral network were obviously blurred and disordered,while in Prednisolone group and Jianpi Yanggan Decoction group,the structure of hepatic sinusoids,and fenestral network were relatively clear and distinct.Spleen and peripheral blood Th17 and Treg cells:Compared with the control group,the model group showed a significant increase in Th17 cells in the spleen and peripheral blood,a significant decrease in Treg cells in peripheral blood(P<0.05),and a significant increase in the ratio of Th17/Treg cells in the spleen(P<0.01).Compared with model group,the spleen Th17 cells and Th17/Treg cell ratio were significantly decreased in Prednisolone group and Jianpi Yanggan Decoction group(P<0.01).Comparison of plasma inflammatory cytokines:Compared with the control group,the model group showed a significant increase in IL-17 level(P<0.05),and a significant decrease in IL-10 level(P<0.05).Compared with the model group,Jianpi Yanggan Decoction group showed a significant decrease in IL-17 level(P<0.05).Immunohistochemistry results:Compared with the control group,the model group showed an increase in RORγt expression.Compared with the model group,the expression of RORγt was reduced in Prednisolone group and Jianpi Yanggan Decoction group.Conclusion Jianpi Yanggan Decoction may improve the structure of hepatic sinusoids and the immune microenvironment of liver sinuysoidal endothelial cells by regulating the expression of transcription factors ROR γt and Foxp3,regulating the balance of Th17/Treg axis,reducing the expression of inflammatory factors,and thereby alleviating liver inflammatory damage.
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