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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。
OBJECTIVE: To observe the balance of Th17 / Treg in rat model of damp-heat colitis and the regulation of Th17 / Treg by Huangqin Decoction, and to explore its mechanism of treatment for damp-heat colitis from immunological point of view. Methods: The rat model of damp-heat was established by compound factors such as high-fat and high-sugar diet combined with artificial climate chamber method. Combined with trinitrobenzene sulfonic acid (TNBS) enema, damp heat + TNBS colitis model was established. Sixty rats Randomly divided into blank group, damp heat + TNBS colitis model group, Huang Qin Decoction group, Mesalazine group. The rats were subjected to general clinical observation. The levels of IL-17, IL-6 and IL-10 in serum were determined by enzyme-linked immunosorbent assay (ELISA) ) And transforming growth factor β (TGF-β) were detected by immunohistochemistry. The expression of FOXP3 and Th17 transcription factor RORγt in colon were detected by Western blot. Results: Compared with the control group, the anti-inflammatory effect of Huangqin Tang decoction 0.12g / 100g could effectively relieve the inflammatory reaction of Th17 / Treg in rats with damp-heat and TNBS-induced colitis, the Th17 cytokine IL-17 The expression of IL-6, RORγt and Treg cytokines IL-10 and TGF-β and FOXP3 were significantly decreased. Compared with model group, Huangqin Decoction could reduce the expression of IL-17, IL-6 and RORγt Level to enhance the expression of IL-10, TGF-β and FOXP3. CONCLUSION: The imbalance of Th17 / Treg may be closely related to the development of damp-heat colitis. Scutellaria decoction can effectively regulate the balance of Th17 / Treg, which may be one of its immune mechanisms in the treatment of damp-heat colitis.