| Background:Ginseng plays a role in cardiovascular,endocrine and other systems,and also has a certain regulatory role in the immune system.AD-1 is a saponin derivative extracted from the berries of ginseng.It has been proved to have a significantly higher inhibitory effect on lung cancer cells than other saponins,but its regulation on immune cells is rarely studied.T cells are classified into CD4~+T cells and CD8~+T cells according to their recognition of antigens.CD4~+T cell subsets can differentiate into T helper 1,Th1,Th2,Th9,Th17,Th22,follicular helper T cells(Tfh)and regulatory T cells(Treg)and other different cells.The body’s inflammation is usually accompanied by the disorder of the CD4~+T cell subsets in the body.Studies have confirmed that the CD4~+T cell subsets in the body are disordered after inflammatory bowel disease(IBD).However,the current drugs used for its treatment generally have disadvantages such as large side effects and recurrence of the disease.Traditional Chinese medicine treatment has become a new treatment method.The search for new drugs that can regulate T cell differentiation is of great significance for the treatment of IBD.Whether AD-1 can improve IBD through CD4~+T cell subsets has not been reported.Therefore,we used AD-1 to act on IBD mice to explore whether it can improve chronic IBD in mice by regulating changes in CD4~+T cell subsets.Objective:Explore whether AD-1 can regulate CD4~+T cell subsets in vivo to improve chronic IBD in mice;induce differentiation of mouse CD4~+T cells in vitro,and explore the effects of AD-1 on cell activation,proliferation and differentiation.Methods:AD-1 can improve the colon shortening caused by chronic IBD in mice induced by DSS,and at the same time can reduce the mouse DAI score;the results of HE staining of colon tissue sections show that the model group mouse colon mucosa is seriously damaged,the epithelial cell structure is disordered,and there is hidden Fossa abscess,goblet cells disappeared,and a large number of inflammatory cells infiltrated;the colon tissue structure of the AD-1 group was relatively complete,which significantly improved the structural disorder of colon epithelial cells,reduced inflammatory cell infiltration,destroyed the crypt structure and disappeared goblet cells The situation has improved;ELISA method detects the expression of IFN-γ,IL-4,IL-17A,IL-10 cytokines in the colon of mice,and the results show that AD-1significantly reduces the levels of IFN-γ,IL-17A Secretion,significantly up-regulate the secretion of IL-4 and IL-10;WB method detects mouse colon tissue and found that AD-1 significantly down-regulates the expression of T-bet and RORγt in the colon,and up-regulates the expression of GATA3 and Fox P3;IHC staining results Compared with the Model group,the colon tissues of the AD-1 group had significantly higher expressions of GATA3 and Fox P3 positive results,while the expression of T-bet and RORγt positive results were significantly reduced;the results of flow cytometry showed that the AD-1 group was compared with the Model group.Compared with the group,it can inhibit the differentiation of Th1 and Th17 cells in mouse spleen and MLN,and can promote the differentiation of Th2 and Treg cells.CCK-8 results showed that AD-1(5,10,20μM)had no effect on the proliferation of Naive CD4~+T cells sorted from mouse spleen;flow cytometry results showed that AD-1(5,10,20μM)had an effect on CD4~+The expression of T cell activation marker CD69 has no effect.AD-1 can inhibit the differentiation of Th1 and Th17 cells in vitro,and promote the differentiation of Th2 and Treg cells in vitro.Results:AD-1 can improve the colon shortening caused by chronic IBD in mice induced by DSS,and reduce the DAI score of mice.The results of HE staining of colon tissue sections show that the mice in the Model group have serious colonic mucosa damage,disordered epithelial cell structure,and appearance Fossa abscess,goblet cells disappeared,and a large number of inflammatory cells infiltrated;the colon tissue structure of the AD-1 group was relatively complete,which significantly improved the structural disorder of colon epithelial cells,reduced inflammatory cell infiltration,destroyed the crypt structure and disappeared goblet cells The situation has improved;ELISA method detects the expression of IFN-γ,IL-4,IL-17A,IL-10cytokines in the colon of mice,and the results show that AD-1 significantly reduces the levels of IFN-γ,IL-17A Secretion,significantly up-regulate the secretion of IL-4and IL-10;WB method detects mouse colon tissues and found that AD-1 significantly down-regulates the expression of T-bet and RORγt in the colon,and up-regulates the expression of GATA3 and Fox P3;IHC staining Compared with the Model group,the colon tissues of the AD-1 group had significantly higher expressions of GATA3 and Fox P3 positive results,while the expression of T-bet and RORγt positive results were significantly reduced;the results of flow cytometry showed that the AD-1 group was compared with the Model group.Compared with the group,it can inhibit the differentiation of Th1 and Th17 cells in mouse spleen and MLN,and can promote the differentiation of Th2 and Treg cells.CCK-8 results showed that AD-1(5,10,20μM)had no effect on the proliferation of Naive CD4~+T cells sorted from mouse spleen;flow cytometry results showed that AD-1(5,10,20μM)had an effect on CD4~+The expression of T cell activation marker CD69 has no effect.AD-1 can inhibit the differentiation of Th1 and Th17 cells in vitro,and promote the differentiation of Th2and Treg cells in vitro.Conclusion:1.AD-1 alleviates the inflammation degree of chronic IBD in mice induced by DSS;2.The therapeutic effect of AD-1 on chronic IBD in mice is related to down-regulation of Th1 and Th17 cells and up-regulation of Th2 and Treg cell differentiation;3.In vitro AD-1 does not participate in the activation and proliferation stage of CD4~+T cells,but directly regulates its differentiation stage.AD-1 inhibits the differentiation of Th1 and Th17 cells and promotes the differentiation of Th2 and Treg cells. |
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