艾灸对克罗恩大鼠结肠RoRγt、Foxp3调节作用研究

Study on Regulation Function of Moxibustion on Expressions of Ro Rγt and Foxp3 in Colon of Rats with Crohn Disease

目的:观察隔药灸对克罗恩大鼠结肠Th17特异性转录因子维甲酸相关孤核受体γt(Ro Rγt)和Treg特异性转录因子叉头蛋白P3(Foxp3)的表达调节作用,探讨灸法治疗克罗恩病的可能效应机制。方法:SPF级雄性SD大鼠40只,完全随机分为4组,每组10只:正常组、模型组、隔药灸组和西药组(美沙拉嗪),采用三硝基苯磺酸(TNBS)溶液灌肠制备大鼠克罗恩疾病模型,隔药灸组取穴:天枢(双)、气海穴,西药组:美沙拉嗪灌胃治疗。治疗干预结束后,采用HE病理染色、免疫组织化学和Real-time PCR方法观察大鼠结肠降部组织病理学变化、Foxp3 m RNA的表达情况。结果:模型大鼠结肠组织出现克罗恩病的典型病理表现:裂隙状溃疡、黏膜层炎症伴肉芽肿,Th17特异性转录因子Ro Rγt蛋白在模型大鼠结肠黏膜及黏膜下层表达高于正常组大鼠(P<0.01),隔药灸和美沙拉嗪对Ro Rγt蛋白具有下调作用(P<0.01),且美沙拉嗪的调节作用强于隔药灸(P<0.01);Treg细胞特异性转录因子Foxp3及其m RNA在模型大鼠结肠黏膜及黏膜下层表达均低于正常组大鼠(P<0.01),隔药灸和美沙拉嗪对Foxp3及其m RNA都具有上调作用(P<0.01),但两组间比较无统计学差异(P>0.01)。结论:隔药灸可能通过调节克罗恩病模型大鼠结肠黏膜Th17/Treg细胞平衡中特异性转录因子Ro Rγt、Foxp3的表达来发挥改善结肠组织溃疡,缓解肠道炎症的治疗作用。

This study was aimed to observe the regulation function of herbs-partition moxibustion on the expressions of Ro Rγt and Foxp3 in the colon of rats with crohn disease（CD）, and to explore the mechanism of moxibustion in the treatment of CD. A total of 40 male SD rats of specific pathogen free grade were randomized into the normal group, the model group, the herbs-partition moxibustion group and the west medicine group（mesalazine）, with 10 rats in each group. Trinitro-benzene-sulfonic acid（TNBS） was used to develop CD models. Tianshu（ST25, both sides） and Qihai（RE6） were selected in herbs-partition moxibustion group. In the west medicine group, rats received mesalazine by gavage. At the end of the intervention, histopathological changes and Foxp3 m RNA expression in rats＇ descending colon were detected by using HE pathological dyeing, immunohistochemistry and real-time PCR. The results showed that colon tissues of rats in the model group expressed typical pathologic signs of CD： crack ulcer and mucosa inflammatory granuloma. The protein expression of Ro Rγt in the model rat colonic mucosa and submucosa was higher than that of the normal group（P〈 0.01）. Both herbs-partition moxibustion and mesalazine can decrease the protein expression of Ro Rγt（P〈 0.01）. The regulation function of mesalazine was stronger than that of herbs-partition moxibustion（P〈 0.01）. The protein expression of Foxp3 and m RNA in the model rat colonic mucosa and submucosa was lower than that of the normal group（P〈 0.01）. Both herbs-partition moxibustion and mesalazine can lift the protein expression of Foxp3 and m RNA（P〈 0.01）, but there was no statistical difference between two groups（P〉 0.01）. It was concluded that the possibly mechanisms of herbs-partition moxibustion to improve the colon tissue ulcers and alleviate the intestinal inflammation were through the regulation on the expression of Ro Rγt and Foxp3 in the CD model rat colonic mucosa Th17/Treg cell.