溃结宁膏穴位敷贴对溃疡性结肠炎（脾肾阳虚证）大鼠TLR4/MyD88/NF-κB信号通路的影响

Effect of Kuijiening Plaster Point Application on TLR4/MyD88/NF-κB Signal Pathway in Ulcerative Colitis Rats with Spleen-kidney Yang Deficiency

【目的】观察溃结宁膏穴位敷贴对溃疡性结肠炎(脾肾阳虚证)大鼠Toll样受体4(TLR4)/髓样分化因子88(MyD88)/核转录因子kappaB(NF-κB)信号通路的干预作用,探讨其治疗溃疡性结肠炎的作用机制。【方法】将40只SD大鼠随机分为正常组、模型组、敷贴组、柳氮磺吡啶(SASP)组,每组10只。采用番泻叶、腺嘌呤分阶段灌胃结合外因干预并给予2,4,6-三硝基苯磺酸(TNBS)与乙醇复合物诱导灌肠法复制溃疡性结肠炎动物模型。造模结束后1周,敷贴组大鼠开始给予溃结宁膏穴位敷贴,SASP组给予SASP灌胃,每日1次,共28 d。采用免疫组化法和逆转录聚合酶链反应(RT-PCR)法分别检测各组大鼠结肠组织TLR4、MyD88、NF-κB蛋白与mRNA的表达。【结果】与正常组比较,模型组、敷贴组、SASP组大鼠TLR4、MyD88、NF-κB蛋白与mRNA表达水平均显著升高(P<0.01);与模型组比较,敷贴组与SASP组大鼠TLR4、MyD88、NF-κB蛋白与mRNA表达水平均明显降低(P<0.01),但2组组间差异无统计学意义(P>0.05)。【结论】溃结宁膏穴位敷贴可有效治疗大鼠溃疡性结肠炎(脾肾阳虚证),其作用机制可能与其阻断TLR4/MyD88/NF-κB信号通路的转导,从而减轻肠道炎症反应有关。

Objective To observe the intervention effect of Kuijiening Plaster point application on Toll-like receptor 4(TLR4)/myeloid differentiation factor 88(MyD88)/nuclear factor kappaB(NF-κB)signaling pathway in ulcerative colitis(UC)rats with spleen-kidney yang deficiency,and to explore its possible mechanism. Methods Forty Sprague-Dawley(SD) rats were randomized into normal group, model group, application group and sulfasalazine(SASP)group,10 rats in each group. The rat model was established by gavage of the decoction of Folium Sennae and adenine at different stages combined with external intervention as well as enema with the compound of 2,4,6-trinitrobenzene sulfonic acid(TNBS)and alcohol. One week after the modeling ended,the application group was given Kuijiening Plaster point application, and SASP group was given intragastric administration of SASP,once a day,the treatment lasting 28 d. And then the protein and mRNA expression levels of TLR4, MyD88 and NF-κB in colonic tissues of rats were detected by immunohistochemistry and reverse transcription polymerase chain reaction(RT-PCR)method, respectively. Results Compared with the normal group,the protien and m RNA expression levels of TLR4,MyD88 and NF-κB in the model group,application group and SASP group were significantly increased(P < 0.01). Compared with the model group,the protein and mRNA expression levels of TLR4,MyD88 and NF-κB in rats were significantly decreased in the application group and SASP group(P < 0.01),but there was no significant difference in the both groups(P > 0.05). Conclusion Kuijiening Plaster point application has obvious effect on improving UC with spleen-kidney yang deficiency. Its mechanism may be related with inhibiting the transduction of TLR4/MyD88/NF-κB signaling pathway which resuls in the relief of the intestinal inflammatory reaction.