Study On The Role Of LncRNA XIST In Immune Cells Of Primary Biliary Cholangitis

Objective:Primary biliary cholangitis(PBC)is an autoimmune disease characterized by multiple female cholangitis and liver damage.The significant difference between male and female is a starting point to explore the pathogenesis of PBC.In this study,differences of LncRNA XIST in immune cells of patients with PBC and healthy control group were investigated,as well as the effects of LncRNA XIST on na?ve CD4~+T lymphocytes proliferation,typing and related cytokines in patients with PBC.Methods:PBC and normal peripheral venous blood samples were collected,immune cells in peripheral venous blood were separated by magnetic beads,including NKs cells,B cells,CD4~+T cells,CD8~+T cells,using real-time fluorescent quantitative(qRT-PCR)and fluorescence in situ hybridization(FISH)quantitative analysis and positioning PBC patients and normal immune cells LncRNA XIST expression level.Then,siRNA was used to screen out appropriate sequences,lentivirus was used to stabilize the expression of LncRNA XIST,and CCK8 method was used to detect the effect of LncRNA XIST on Na?ve CD4~+T cells proliferation in PBC patients.Finally,na?ve CD4~+T cells interfering with the expression of LncRNA XIST were co-cultured with bile duct epithelial cells,and the effects of LncRNA XIST on the typing of na?ve CD4~+T cells and related cytokines were detected by qRT-PCR,enzyme-linked immunoadsorption assay(ELISA)and flow cytometry(FACS).Results:The expression levels of LncRNA XIST of various immune cells in peripheral venous blood of patients with PBC and healthy control group were detected by q RT-PCR.It was found that the expression levels of LncRNA XIST in NK and CD4~+T lymphocytes in PBC patients were significantly higher than those in healthy control group(1.19±?1.23 vs 0.34±?0.31,p<0.01;0.48±?0.19 vs 1.26±?0.09,p<0.05),and could be clearly located on the nucleus.Lentivirus was used to interfere with the expression of LncRNA XIST in na?ve CD4~+T cells in PBC patients.It was found that LncRNA XIST could promote the proliferation of na?ve CD4~+T cells.Through co-culture of na?ve CD4~+T cells with bile duct epithelial cells,it was found that interfering with the expression of LncRNA XIST could reduce the secretion of IFN-?,IL-17,T-bet and ROR?t in na?ve CD4~+T cells,thereby reducing the proportion of Th1 and Th17 cells(5.88±1.18 vs 2.64±0.79,p<0.01;4.91?±?1.51 vs 2.3?±?0.76,p<0.05).Conclusion:LncRNA XIST was associated with lymphocyte abnormalities in patients with PBC.The high expression of LncRNA XIST in na?ve CD4~+T cells in PBC patients stimulates na?ve CD4~+T cell proliferation and stimulates the secretion of inflammatory cytokines,leading to the increase of Th1 and Th17 ratios,which may contribute to the occurrence of PBC.Therefore,this study provides a direction for further exploration of the pathogenesis of PBC and the search for new targets for clinical drug therapy.