丁氏溃结灌肠液对溃疡性结肠炎模型大鼠IL-22/STAT3通路的影响

Influence of Ding's Kuijie enema on IL-22/STAT3 signaling pathway in DSS-induced rat with ulcerative colitis

目的:观察丁氏溃结灌肠液对葡聚糖硫酸钠(DSS)模型大鼠血清IL-22/STAT3信号通路的影响,探讨其改善溃疡性结肠炎肠道稳态的作用机制。方法:60只8周龄健康大鼠,分为空白组、模型组、丁氏溃结灌肠液组6 g/kg、3 g/kg、1. 5 g/kg组、美沙拉秦2. 4 g/kg组。除空白组,其他各组分别饮水喂食3. 5%葡聚糖硫酸钠7天,进行药物灌肠治疗7天。酶联免疫法(ELISA)检测各组组织中IL-1β的含量及血浆中IL-22的含量;实时定量PCR检测各组结肠组织SOCS3 mRNA的表达水平;免疫印迹(Western blot)检测各组STAT3、磷酸化STAT3(p-stat3)及下游目标蛋白SOCS3的表达水平;免疫组化染色Ki-67评估各组结肠组织的增殖活性及状态。结果:与空白组比较,模型组IL-1β、IL-22含量显著升高,p-stat3蛋白表达水平显著升高,SOCS3蛋白表达水平无差异;SOCS3 mRNA表达水平显著下降,Ki-67染色阳性率显著上升;与模型组比较,丁氏溃结灌肠液组6 g/kg、3 g/kg、1. 5 g/kg组及美沙拉秦2. 4 g/kg组IL-1β、IL-22含量显著下降,p-stat3蛋白表达水平下降,ki-67染色阳性率显著下降。结论:丁氏溃结灌肠液改善溃疡性结肠炎的肠道稳态作用可能与提高IL-22含量,激活stat3通路,促进肠上皮细胞增殖有关。

Objective： To observe the influence of Ding’s Kuijie enema on IL-22/STAT3 signaling pathway in Dextran Sulfate Sodium（DSS）-induced rats with experimental ulcerative colitis,and explore its underlying mechanism of intestinal stability. Methods： 60 rats were randomly selected as the control group,the model group,the low,medium and high dose group,and the mesalazine enema group. Except the control group,each group were given 5% DSS 7 days and drugs for 7 days corresponding,the control group free feeding. colon specimens to observe the number of colon crypts,determined the content of IL-1βin colon tissue and IL-22 in plasma by ELISA,Wstern Blot was used to detect the protein expression levels of stat3,p-stat3 and downstream target SOCS3. Real-Time PCR was used to detect the expression levels of SOCS3 mRNA in the colon tissue,Immunohistochemical was used to detect the positive rate of Ki-67,evaluating the proliferative activity and status of colon tissue in each group. Results： Compared with the control group,in the model group,the content of IL-1β、IL-22 increased significantly（P 〈 0. 01）,the protein expression of p-stat3 increased and socs3 decreased,the mRNA expression of SOCS3 decreased significantly（P 〈 0.01）. the positive rate of Ki-67 increased significantly（P 〈 0. 01）. Compared with the model group,above indexes were ameliorated in Ding’s Kuijie enema groups and mesalazine enema group,and there was significant difference compared with the model group（P 〈 0. 01）. Conclusion： Ding’s Kuijie enema can significantly alleviate the symptoms of UC rats,there have a protective effect on the intestinal stability,The possible mechanism is Ding’s Kuijie enema can improve the content of IL-22,activate stat3 pathway,and promote intestinal epithelial cell proliferation.