针灸治疗对狼疮性肾炎小鼠Th17/Treg细胞及其细胞因子表达影响的研究

目的:观察针灸治疗对狼疮性肾炎小鼠的影响,并探讨针灸对调节性T细胞(Treg)及辅助性T细胞17 (Th17)分化及其细胞因子表达的调控作用。方法:将MRL/lpr狼疮性肾炎模型小鼠随机分为针灸组、地塞米松组、模型组,每组10只。另取10只C57BL/6小鼠作为正常组。采用HE染色法检测小鼠肾脏病理改变;PI单染流式细胞术检测各组小鼠肾脏组织中Th17/Treg比例;采用q RT-PCR和ELISA法检测肾脏中叉头样转录因子3 (Foxp3)、维A酸相关孤儿受体-γt (RORγt)及白细胞介素-17 (IL-17)的水平。结果:与正常组比较,模型组肾小球肾炎、间质性肾炎以及血管病变评分、Th17/Treg比值及RORγt、IL-17蛋白及mRNA表达水平均显著升高(P<0.05),Foxp3蛋白及mRNA表达水平均下降(P<0.05)。与模型组比较,地塞米松组、针灸组肾小球肾炎、间质性肾炎以及血管病变评分、Th17/Treg比值及肾脏组织RORγt、IL-17蛋白及m RNA表达水平均显著降低(P<0.05);Foxp3蛋白及mRNA表达均升高(P<0.05)。结论:针灸能显著减少狼疮性肾炎小鼠肾脏RORγt及IL-17表达,增加Foxp3水平,调控Th17/Treg比例,改善肾脏病理改变,从而对MRL/lpr狼疮肾炎小鼠肾脏起到保护作用。

Study on Acupuncture Treatment has Effect on Th17/Treg Cells and the Cytokine Expression in Mice with Lupus Nephritis

Objective:To observe the effect of acupuncture on mice with lupus nephritis,and to discuss its regulatory effect on differentiation of regulatory T cell(Treg) and T helper 17 cell(Th17) as well as their cytokine expression. Methods:MRL/lpr mice models with lupus nephritis were randomly divided into the acupuncture group,the dexamethasone group and the model group, 10 mice in each group. Another 10 C57 BL/6 mice were selected as the normal group. The pathological changes of the kidney were detected by HE staining;the ratio of Th17/Treg in the kidney tissue in each group was detected by PI single-staining flow cytometry;the levels of forkhead box protein 3(Foxp3),retinoic acid-related orphan receptor-γt(RORγt) and interleukin-17(IL-17) in the kidney were detected by qRT-PCR and ELISA method. Results:Compared with those in the normal group,the scores of glomerulonephritis,interstitial nephritis and angiopathy,the ratio of Th17/Treg and the level of expression of RORγt and IL-17 potein as well as their mRNA in the model group were significantly increased(P<0.05),and the level of expression of Foxp3 protein and its mRNA was decreased(P<0.05). Compared with those in the model group, the scores of glomerulonephritis, interstitial nephritis and angiopathy, the ratio of Th17/Treg and the level of expression of RORγt and IL-17 protein as well as their mRNA in the dexamethasone group and the acupuncture group were significantly decreased(P<0.05), and the level of expression of Foxp3 protein and its mRNA was increased(P<0.05).Conclusion: Acupuncture can significantly decrease the expression of RORγt and IL-17 in the kidney of mice with lupus nephritis,increase the level of Foxp3,regulate the ratio of Th17/Treg and improve the pathological changes of the kidney,thus protecting the kidney of MRL/lpr mice with lupus nephritis.