河北医学2024，Vol.30Issue(8)：1267-1272,6.DOI:10.3969/j.issn.1006-6233.2024.08.07
BAFF调节免疫性血小板减少症模型小鼠的Th17/Treg平衡的研究
Regulation of Th17/Treg Balance by BAFF in a Mouse Model of Immune Thrombocytopenia
摘要
Abstract
Objective:To explore the regulatory role of B-cell activating factor(BAFF)on the balance of helper T cell 17(Th17)and regulatory T cell(Treg)in vivo in a mouse model of immune thrombocytope-nia(ITP).Methods:Guinea pig anti-mouse platelet serum(GP-APS)was prepared,and 150 adult male BALB/c mice(7-8 weeks old)were randomly divided into 5 groups(n=30 per group):control(blank con-trol),ITP(induced by GP-APS),ITP+rhBAFF(ITP mice treated with intravenous injection of recombinant human BAFF protein at 50μg/kg/50μL),ITP+rhBAFF+DAPT(combined with Notch1 inhibitor DAPT),and ITP+rhBAFF+PGD(combined with PI3K/Akt inhibitor Polygalacin D).DAPT was injected at 100μg/kg,and PGD at 25 μg/kg,both with a total volume of 50μL and administered once daily.Peripheral blood(1mL)was collected from mice after 1 week for serum and mononuclear cell isolation.Immunofluorescence staining was used to detect BAFF and Notch1 localization in mononuclear cells.Platelet counts in peripheral blood were determined.Enzyme-linked immunosorbent assay(ELISA)was performed to measure BAFF lev-els in mouse serum.Western blotting was conducted to assess protein expression of PI3K,AKT,Notch1,p-Akt(Thr308),and p-Akt(Ser473)in mononuclear cells.Flow cytometry was used to analyze changes in Th17/Treg ratio in mononuclear cells.Results:BAFF and Notch1 were co-localized on the cell membrane of mononuclear cells in peripheral blood of ITP mice.Compared with the control group,ITP group showed in-creased expression of BAFF,Notch1,p-Akt(Thr308),p-Akt(Ser473),increased Th17 proportion,de-creased platelet count,and decreased Treg proportion(P＜0.05).Compared with the ITP group,ITP+rh-BAFF group exhibited increased BAFF,Notch1,p-Akt(Thr308),p-Akt(Ser473)expression,increased Th17 proportion,decreased platelet count,and decreased Treg proportion(P＜0.05).Compared with the ITP+rhBAFF group,both ITP+rhBAFF+DAPT and ITP+rhBAFF+PGD groups showed decreased BAFF,Notch1,p-Akt(Thr308),p-Akt(Ser473)expression,increased platelet count,increased Treg proportion,and de-creased Th17 proportion(P＜0.05).Conclusion:BAFF promotes an increase in Th17 proportion and de-crease in Treg proportion in a mouse model of immune thrombocytopenia by activating the Notch1/PI3K/Akt signaling pathway.关键词
B细胞激活因子/免疫性血小板减少症/小鼠/辅助性T细胞17/调节性T细胞的平衡Key words
B-cell activating factor/Immune thrombocytopenia/Mouse/Th17/regulatory T cell balance引用本文复制引用
李巍,马西虎,刘晓,费飞,秦兰,买尔吾甫·木合布力,白玉盛..BAFF调节免疫性血小板减少症模型小鼠的Th17/Treg平衡的研究[J].河北医学,2024,30(8):1267-1272,6.基金项目
新疆维吾尔自治区自然科学基金项目,(编号:2022D01C167) （编号:2022D01C167）
