温阳化浊通络方对系统性硬化病患者Th17细胞增殖的影响

Objective: To study the effect of the Wenyang Huazhuo Tongluo Formula on the proliferation of Th17 cells in patients with systemic sclerosis. Methods: Peripheral blood mononuclear cells (PBMC) were isolated from patients by density gradient centrifugation. Th17 cells in PBMC were screened by the Stemcell Th17 magnetic sorting kit, and the drug-containing serum of the Wenyang Huazhuo Tongluo Formula was prepared simultaneously. Before and after Th17 cells were transfected with miR - 155 agomir and miR - 155 antagomir respectively, the effects of the drug-containing serum on the expressions of miR - 155, IL - 6 and IL - 6R mRNA in Th17 cells were detected by RT - qPCR, and the effects of the drug-containing serum on the expressions of IL - 6 and IL - 6R proteins in Th17 cells were detected by cell immunofluorescence technique. Results: Compared with the blank serum group, the drug-containing serum could inhibit the proliferation index of Th17 cells and the expression of miR - 155 in a dose-dependent manner (P0.05), but it could still inhibit the expressions of IL - 6 mRNA and protein in a dose-dependent manner (P < 0.01). Conclusion: The Wenyang Huazhuo Tongluo Formula can inhibit the proliferation of Th17 cells in patients with systemic sclerosis. Its mechanism may be related to the inhibition of the expressions of miR - 155, IL - 6 and IL - 6R mRNA and proteins, and the inhibitory effect on the latter may also be related to the direct regulation of miR - 155.

目的研究温阳化浊通络方对系统性硬化病患者Th17细胞增殖的影响。方法密度梯度离心法分离患者外周血单核细胞( PBMC),Stemcell Th17磁性分选试剂盒筛选PBMC中Th17细胞,同时制备温阳化浊通络方含药血清。 Th17细胞分别转染miR-155 agomir、miR-155 antagomir前后,RT-qPCR检测含药血清对Th17细胞中miR-155、IL-6、 IL-6R mRNA表达的影响,细胞免疫荧光技术检测含药血清对Th17细胞中IL-6、IL-6R蛋白表达的影响。结果与空白血清组比较,含药血清可剂量依赖性地抑制Th17细胞增殖指数和miR-155表达( P0.05),但仍可剂量依赖性地抑制IL-6 mRNA和蛋白表达( P<0.01) 。结论温阳化浊通络方可抑制系统性硬化病患者Th17细胞增殖,其机制可能与抑制miR-155及IL-6、IL-6R mRNA和蛋白表达有关,而且对后者的抑制作用还可能与直接调控miR-155有关。