芪连结肠宁对溃疡性结肠炎大鼠TLR4/NF-κB信号通路及肠黏膜保护因子的影响

Effect of Qilian Jiechang Ning on the TLR4/NF-κB signaling pathway and intestinal mucosal protective factors in ulcerative colitis rats

目的研究芪连结肠宁对溃疡性结肠炎大鼠TLR4/NF-κB信号通路及肠黏膜保护因子的影响。方法健康雄性SD大鼠50只,随机分为正常组、模型组、芪连结肠宁低、中、高剂量组,每组10只。采用5%葡聚糖硫酸钠(DSS)喂养建立溃疡性结肠炎大鼠模型,连续给药3周后检测指标。观察体质量变化及DAI评分;HE染色观察结肠组织病理情况;免疫组化法观察结肠组织MUC2表达情况;Western blot检测结肠组织TLR4、NF-κB p65、IL-6、TNF-α、MUC2、TFF3蛋白表达水平;实时荧光定量PCR检测结肠组织MUC2、TFF3 mRNA的表达。结果芪连结肠宁各组大鼠在给药之后体质量明显增加(P<0.01),DAI评分明显降低(P<0.01),且中、高剂量组具有明显优势。芪连结肠宁各组大鼠结肠组织病理损伤不同程度恢复,MUC2、TFF3蛋白表达相比于模型组均明显增加(P<0.01)。与模型组比较,芪连结肠宁各组大鼠结肠组织TLR4、p-NF-κB p65、IL-6、TNF-α蛋白表达均有不同程度的下降(P<0.05,P<0.01),TFF3 mRNA相对表达量显著增加(P<0.01)。结论芪连结肠宁能有效恢复溃疡性结肠炎模型大鼠结肠组织的病理损伤,降低DAI评分,降低结肠组织TLR4、p-NF-κB p65、IL-6、TNF-α蛋白表达,增加MUC2、TFF3蛋白及TFF3 mRNA的表达,进而治疗溃疡性结肠炎,其中芪连结肠宁中、高剂量组疗效及调节作用较为显著。

Objective To study the effect of Qilian Jiechang Ning on the TLR4/NF-κB signaling pathway and intestinal mucosal protective factors in ulcerative colitis rats.Methods A total of fifty healthy male SD rats were randomly divided into a normal group,a model group,and low-,medium-and high-dose Qilian Jiechang Ning groups,with 10 rats in each group.A rat model of ulcerative colitis was established by feeding them with 5%DSS,and the indexes were detected after continuous administration for 3 weeks.Changes in body weight and DAI scores were observed;HE staining was used to observe the pathological conditions of colon tissues;immunohistochemistry was used to observe the expression of MUC2 in colon tissues;Western blotting was used to detect the protein expression levels of TLR4,NF-κB p65,MUC2,IL-6 and TNF-αin colon tissues;real-time fluorescence quantitative PCR was used to detect the expression of MUC2 and TFF3 mRNA in colon tissues.Results After the administration of Qilian Jiechang Ning,the body weights of rats in each group were significantly increased(P<0.01),while the DAI scores were significantly decreased(P<0.01),and the middle-and high-dose groups had obvious advantages.Compared with the model group,the pathological damages of colon tissues in the Qilian Jiechang Ning group were recovered to different degrees,and the protein expression levels of MUC2 and TFF3 were significantly increased(P<0.01).Compared with the model group,the protein expression levels of TLR4,p-NF-κB p65,IL-6 and TNF-αin the colon tissues of the Qilian Jiechang Ning groups were decreased to varying degrees(P<0.05,P<0.01),while the relative expression levels of TFF3mRNA were increased significantly(P<0.01).Conclusion Qilian Jiechang Ning can effectively restore the pathological damages of colon tissues in the UC model rats,reduce the DAI scores,reduce the protein expression levels of TLR4,p-NF-κB p65,IL-6,TNF-αin colon tissues,increase the protein expression levels of MUC2,TFF3 and TFF3 mRNA,and then promote the treatment of UC.Among them,the medium-and high-dose Qilian Jiechang Ning groups have a more significant curative effect and regulatory effect.