益气解毒方干预实验性结肠炎大鼠肠上皮细胞自噬功能的研究

Effects of Yiqi Jiedu Formula Interfering Autophagy Function of Intestinal Epithelial Cells of Experimental Colonitis Rats

目的:通过检测益气解毒方对结肠菌株所致实验性结肠炎大鼠肠黏膜上皮细胞自噬相关基因(ATG)16L1及微管相关蛋白轻链3(LC3)的表达及黏膜核因子kappaβ(NF-κβ)活化的影响,探讨益气解毒方防治溃疡性结肠炎的作用机制。方法:构建结肠细菌菌株诱导实验性结肠炎模型,以益气解毒方治疗14 d,设以生理盐水灌胃的模型对照组及空白对照组,观察疾病活动指数(DAI),进行组织学评分(HPS)和分离ICEs并提取ICE中的总RNA,用反转录酶-聚合酶链反应(RT-PCR)法检测ATG16L1及LC3的表达;采用免疫组织化学方法检测NF-κβ的活性情况。结果 :益气解毒组的症状、组织损害程度较模型对照组减轻,其ATG16L1、LC3的表达均高于模型对照组(P<0.05);与空白对照组比较,模型组ATG16L1表达下降,差异有统计学意义(P<0.05);模型对照组NF-κβ活性高于益气解毒组,两组比较差异有统计学意义(P<0.05)。结论:结肠细菌菌株诱导实验性结肠炎大鼠可能存在肠上皮细胞自噬功能不足,益气解毒方通过促使提升上皮细胞自噬功能,进而抑制炎性因子NF-κβ的活性,并在肠道炎症中发挥作用。

Objective：Through detecting Yiqi Jiedu formula on the expression of autophagy related gene16L1（ATG16L1） of epithelial cell of the experimental colonitis rats induced by colon bacteria strains,and effects on microtubule associated protein light chain 3（LC3） and the activation of nuclear factor kappaβ（NF-κβ）,to discuss the mechanism of Yiqi Jiedu formula in treating ulcerative colitis. Methods：We had established the model of the experimental rats with colonitis induced by colon bacteria strains,and used Yiqi Jiedu formula to treat for 14 days,establishing the model control group using normal saline gavage,and blank control group to observe disease activity index（DAI）,make histological score,separate ICEs and extract the total RNA of ICE,using RT-PCR method to detect the expression of ATG16L1 and LC3 and immunohistochemical technique to detect the activity of NF-κβ. Results：The symptom and the degree of tissue damage in the group of Yiqi Jiedu were alleviated significantly than those in the model control group. The expression of ATG16L1,LC3 in the group of Yiqi Jiedu were higher than those in the model control group（P〈0.05）.Comparing with in the blank control group,the expression of ATG16L1 in the model control group decreased,having statistical significance（P〈0.05）. The activity of NF-κβ in the model control group was higher than that in the group of Yiqi Jiedu,having statistical significance（P〈0.05）. Conclusion：The experimental colonitis rats induced by colon bacteria strains may have the deficiency of autophagy function of the intestinal epithelial cells,and Yiqi Jiedu formula can promote the autophagy function of intestinal epithelial cells,so the method can inhibit the activity of inflammatory factor NF-κβ,playing a part in the intestinal inflammation.