他克莫司对小鼠T_H17型细胞分化增殖的影响及其机制

目的 观察在T_H17型细胞极化环境下,他克莫司(Tac)对小鼠T_H17型细胞分化增殖的影响及机制.方法 将小鼠脾脏初始CD4+CD25+T淋巴细胞使用抗CD3抗体及抗CD28抗体进行活化,使用转化生长因子β_1,(TGF-β_1,)和白细胞介素6(IL-6)等细胞因子进行诱导,促使其向T_H17型细胞方向分化.将体外培养的T_H 17型细胞分为4组,分别用不同剂量的Tac进行干预,即:Tac0 ng/ml组(对照组)、Tac 0.1 ng/ml组、Tac 1 ng/ml组和Tae 10 ng/ml组.应用流式细胞术检测各组T_H 17型细胞亚群纯度,使用逆转录聚合酶链反应(RT.PCR)检测各组IL-17 mRNA相对表达量.结果 Tac可抑制小鼠T_H17型细胞亚群的分化增殖,降低IL-17 mRNA的表达量,且呈剂量依赖性,各组间比较,差异均有统计学意义(P<0.05).结论 Tac通过抑制T_H17型细胞胞浆内的钙调磷酸酶,抑制T_H17型细胞亚群的分化增殖,抑制IL-17 mRNA产生并减轻炎症进程,从而抑制排斥反应.

The influonce of tacrolimus on the differentiation and proliferation of T_H 17 cells in mice and the mechanism

Objective To explore the effects of tacrolimus (Tac) on the differentiation and proliferation of T_H 17 (T helper type 17) cells in mice under T_H 17 polarized conditions and the mechanism.Methods Murine naive CD4+ CD25- T cells were stimulated with plate-bound anti-CD3 and anti-CD28, and induced by the cytokines such as TGF-β1 and IL-6, etc.The in vitro T_H17 cells in mice were divided into 4 groups: Tac 0 ng/ml group (control group), Tac 0.1 ng/ml group, Tac 1 ng/ml group and Tac 10 ng/ml group.Tac was added to the cells at the desired time point and desired concentrations.The subsets of CD4+ T_H 17 cells were examined by flow cytometry.The expression of IL-17 mRNA was assayed by RT-PCR Results Tac inhibited the differentiation and proliferation of T_H 17 cells in mice in a dose-dependent manner.The expression of IL-17 mRNA was inhibited either.There were significant difference among the groups (P< 0.05 ).Conclusion By suppression the activation of calcineurin in T_H 17 cells, Tac inhibits the differentiation and proliferation of T_H 17 cells, reduces the expression of IL-17 mRNA and alleviates the process of inflammation, thereby suppressing rejection.