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目的观察溃结灵对溃疡性结肠炎(UC)大鼠模型结肠黏膜调节性T细胞(Treg)相关因子,Foxp3、STAT5的调控作用,探讨溃结灵防治UC的作用机理。方法采用三硝基苯磺酸(TNBS)法复制UC大鼠模型并进行中药复方溃结灵药物干预治疗。采用酶联免疫吸附法(ELISA)检测结肠黏膜白细胞介素2(IL-2)、白细胞介素10(IL-10)的表达,免疫组化方法检测结肠黏膜蛋白原位表达,并采用实时荧光定量PCR(RT-PCR)方法检测Foxp3、STAT5基因表达。结果模型组结肠黏膜IL-2、IL-10表达量均低于正常组(P<0.05,P<0.01),溃结灵高剂量组及阳性药物组柳氮磺胺吡啶(SASP),IL-2表达量高于模型组(P<0.01),溃结灵高、中剂量组及阳性药物组IL-10表达量均高于模型组(P<0.05,P<0.01);免疫组化检测模型组结肠黏膜Foxp3、STAT5的原位蛋白表达低于正常组(P<0.01),溃结灵高、中、低剂量组及阳性药物组Foxp3、STAT5的原位蛋白表达均高于模型组(P<0.05,P<0.01);模型组Foxp3、STAT5 m RNA表达量低于正常组(P<0.05,P<0.01),而溃结灵高、中剂量组及阳性药物组Foxp3 m RNA表达高于模型组(P<0.05,P<0.01),溃结灵高剂量组及阳性药物组STAT5 m RNA高于模型组(P<0.05,P<0.01)。结论溃结灵对UC大鼠模型结肠黏膜IL-2、IL-10含量以及Foxp3、STAT5原位蛋白和基因表达的上调作用,可能是其促进Treg细胞的分化,发挥治疗UC作用的机制之一。
Objective To observe the regulatory effect of Kuijieling on colonic mucosal Treg related factors, Foxp3 and STAT5 in rats with ulcerative colitis (UC) and explore the mechanism of action of Kuijieling on preventing and treating UC. Methods Trinitrobenzene sulfonic acid (TNBS) method was used to replicate the UC rat model and the Chinese medicine compound Kuijieling drug was taken. The expressions of IL-2 and IL-10 in colonic mucosa were detected by enzyme-linked immunosorbent assay (ELISA), the expression of colonic mucosal protein in situ was detected by immunohistochemistry, Quantitative PCR (RT-PCR) was used to detect the expression of Foxp3 and STAT5 genes. Results The expression of IL-2 and IL-10 in colonic mucosa in model group were lower than those in normal group (P <0.05, P <0.01), high dose of Jiegeling group and SASP, IL- (P <0.01). The expression of IL-10 in KUJI group, middle dose group and positive drug group were higher than those in model group (P <0.05, P <0.01). The expression of IL-10 in model group The protein expressions of Foxp3 and STAT5 in colonic mucosa were lower than those in normal group (P <0.01). The protein expressions of Foxp3 and STAT5 in UCK group were higher than those in model group (P < 0.05, P <0.01). The expression of Foxp3 and STAT5 mRNA in model group was lower than that in normal group (P <0.05, P <0.01) (P <0.05, P <0.01). STAT5 mRNA in high-dose Kuijieling group and positive drug group was higher than that in model group (P <0.05, P <0.01). Conclusion Kuiyuling can upregulate the expression of IL-2 and IL-10 in colonic mucosa and the expression of Foxp3 and STAT5 proteins in UC rat model, which may be one of the mechanisms that promote the differentiation of Treg cells and play a role in the treatment of UC .