Th17/Treg平衡在湿热型结肠炎大鼠中的变化及黄芩汤的调节作用

目的:观察湿热型结肠炎大鼠模型中Th17/Treg的平衡变化以及黄芩汤对Th17/Treg的调节作用,从免疫学的角度探讨其治疗湿热型结肠炎的机制。方法:应用高脂、高糖饲料联合人工气候箱法等复合因素造出大鼠湿热模型,再结合三硝基苯磺酸(TNBS)灌肠法建立湿热+TNBS结肠炎模型;将60只大鼠随机分为空白组、湿热+TNBS结肠炎模型组、黄芩汤组、美沙拉嗪组。对大鼠进行一般临床观察,采用酶联免疫吸附法(ELISA)分析大鼠血清中白细胞介素17(IL-17)、白细胞介素6(IL-6)、白细胞介素10(IL-10)和转化生长因子β(TGF-β)水平,用Western blot方法检测结肠组织中Treg转录因子FOXP3和Th17转录因子RORγt的表达水平。结果:黄芩汤免煎剂0.12g/100g能够有效的缓解湿热型结肠炎的炎症反应,与对照组比较,湿热+TNBS结肠炎大鼠模型中Th17/Treg表达失衡,Th17类细胞因子IL-17和IL-6和转录因子RORγt表达升高而Treg类细胞因子IL-10和TGF-β和转录因子FOXP3表达降低;与模型组相比,黄芩汤能够降低IL-17、IL-6和RORγt的水平而增强IL-10、TGF-β和FOXP3的表达。结论:Th17/Treg失衡可能与湿热型结肠炎的发生发展密切相关,黄芩汤能够有效的调节Th17/Treg的平衡关系,这可能是其治疗湿热型结肠炎的免疫机制之一。

The role of Th17/Treg balance in colitis with damp-heat syndrome and the effect of Huangqin Tang

Objective: To clarify the change of Th17 /Treg balance in colitis with damp-heat syndrome and explore the effect of Huangqin Tang on it.Methods: The rat model of colitis with damp-heat syndrome were induced by giving the high sugar fat food and put into an artificial bioclimatic chamber for 4 weeks and then rendered in rats by a single colonic administration of trinitrobenzene sulfonic acid(TNBS).45 rats were randomly divided into three group(n = 3) : damp-heat syndrome + TNBS group,Huangqin Tang treatment group,mesalazine treatment group;another 15 normal rats were used as control group.Assessment of colitis severity was performed daily.The colitis of rats were assessment.cytokines were determined by ELISA.FOXP3 and RORγt were determined by Western blot.Results: Huangqin Tang 0.12g /100g markedly attenuated TNBS-induced colitis with damp-heat syndrome.Th17 / Treg is imbalance in the model rats compared with controls.Th17 markers lilke IL-17,IL-6 and RORγt were up-regulated whereas Treg-related parameters IL-10,TGF-β and FOXP3 were down-regulated.After thera-py,Huang-Qin-Tang ameliorated TNBS-induced colitis with damp-heat syndrome and down-regulated IL-17,IL-6 and RORγt whereas up-regulated IL-10,TGF-β and FOXP3.Conclusion: These data confirm that Huangqin Tang exerted therapeutic effects on colitis with damp-heat syndrome by regulating the banlance of Th17 / Treg.