黄芪甲苷在体外对高迁移率族蛋白B1介导小鼠调节性T淋巴细胞免疫功能的影响

Effects of astragaloside IV on immune function of regulatory T cell in mice mediated by high-mobility group protein B1 in vitro

目的在前期实验基础上,明确经高迁移率族蛋白B1(HMGB1)刺激的小鼠调节性T细胞(Treg)对CD4+CD25-T淋巴细胞免疫功能的影响,观察黄芪甲苷(AST IV)对HMGB1介导Treg免疫功能的拮抗作用。方法随机将CD4+CD25-T细胞分为对照组、Treg组、(HMGB1+Treg)组、(HMGB1+AST IV+Treg)组。4组均于培养后72 h检测脾脏CD4+CD25-T细胞增殖活性及CD4+CD25-T细胞孵育上清液中IL-4、IFN-γ表达水平。结果 Treg组CD4+CD25-T细胞增殖活性受到抑制(P<0.01),其上清中IFN-γ表达水平较对照组明显下降(P<0.01),IL-4表达水平较对照组显著增加(P<0.01);HMGBI+Treg组CD4+CD25-T细胞增殖活性受抑制程度明显逆转(P<0.01),与Treg组比较,(HMGB1+Treg)组IFN-γ表达水平明显升高(P<0.01),IL-4表达水平显著下降(P<0.01);与(HMGB1+Treg)组比较,(HMGB1+AST IV+Treg)组CD4+CD25-T细胞增殖受抑制程度显著加重(P<0.01),IFN-γ表达水平明显降低(P<0.01),IL-4水平显著上升(P<0.01)。结论 HMGB1在体外可明显降低小鼠Treg免疫抑制能力,而黄芪甲苷可拮抗此抑制作用,表明其对HMGB1介导的促炎效应具有治疗作用。

Objective Based previous studies to explore the effects of mice regulatory T cell（ Treg） stimulated by high-mobility group protein B1（ HMGB1） on immune function of CD4^＋CD25^-T cells,and to observe the antagonistic effects of astragaloside IV（ AST IV）on immune function of Treg mediated by HMGB1. Methods CD4^＋CD25^-T cells were randomly divided into a control group,a Treg group,a（ HMGB1 ＋ Treg） group and a（ HMGB1 ＋ AST IV ＋ Treg） group. 72 h after cell culture,CD4^＋CD25^-T cells and the liquid supernatant were again collected to measure the proliferative activity of CD4^＋CD25^-T cells and expression level of IL-4 and IFN-γ in the liquid supernatant in spleen in four groups. Results The proliferative activity of CD4^＋CD25^-T cells was inhibited in the Treg group（ P〈0. 01）,and the expression level of IFN-γ was significantly increased compared with that in the control group（ P〈0. 01）. When Treg that was stimulated with HMGB1 was added in the（ HMGB1 ＋ Treg） group,the inhibited proliferative activity of CD4^＋CD25^-T cells was significantly reversed（ P〈0. 01）,compared with the Treg group,the expression level of IFN-γ in the HMGB1 ＋ Treg group was obviously increased and that of IL-4 was evidently reduced（ P〈0. 01）. Compared with（ HMGB1 ＋ Treg） group,the degree of inhibited proliferative activity of CD4^＋CD25^-T cells was enhanced（ P〈0. 01）,the expression level of IFN-γ was obviously reduced and that of IL-4 was evidently increased（ P〈0. 01）. Conclusion HMGB1 in vitro could obviously reduce Treg immunosuppression in mice,and astragaloside IV have an antagonism effect, indicating its therapeutic effect on pro-inflammatory effects meditated byHMGB1.