优化溃结方对溃疡性结肠炎大鼠结肠Toll样受体/髓样分化因子88/核转录因子κB信号通路的影响

Effect of optimized Kuijie Prescription on colon tissue in ulcerative colitis rats by regulating TLR/MyD88/NF-κB signaling pathway

目的观察优化溃结方对溃疡性结肠炎(UC)大鼠结肠Toll样受体/髓样分化因子88/核转录因子κB(TLR/MyD88/NF-κB)信号通路的影响。方法将40只雄性SD大鼠随机分为空白组、模型组、优化溃结方组、柳氮磺吡啶组,每组10只。除空白组外,其余3组均参考三硝基苯磺酸(TNBS)/乙醇二次致炎法结合束缚法建立UC气滞血瘀型大鼠模型。造模成功后优化溃结方组大鼠予优化溃结方药液1.674 g/(kg·d)灌胃,柳氮磺吡啶组大鼠予柳氮磺吡啶药液0.54 g/(kg·d)灌胃,空白组、模型组不予任何治疗。14天后检测各组大鼠血清白细胞介素17A(IL-17A)、IL-10、IL-22水平,结肠组织MyD88、TLR2、TLR4、NF-κB p65蛋白和mRNA表达水平。结果与空白组相比,模型组血清IL-17A、IL-22水平显著升高(P<0.05),IL-10水平显著下降(P<0.05);与模型组相比,优化溃结方组、柳氮磺吡啶组IL-17A、IL-22水平显著降低(P<0.05),IL-10水平显著升高(P<0.05);优化溃结方组IL-17A、IL-22、IL-10水平与柳氮磺吡啶组比较差异无统计学意义(P>0.05)。与空白组相比,模型组结肠组织MyD88、TLR2、TLR4、NF-κB p65蛋白和mRNA表达均显著增加(P<0.05);与模型组相比,柳氮磺吡啶组、优化溃结方组MyD88、TLR2、TLR4、NF-κB p65蛋白和mRNA表达均显著下降(P<0.05);优化溃结方组MyD88、TLR2、TLR4、NF-κB p65蛋白和mRNA表达与柳氮磺吡啶组比较差异均无统计学意义(P>0.05)。结论优化溃结方可能通过调节TLR/MyD88/NF-κB信号通路,降低炎症因子水平,减轻炎症反应,修复结肠组织的超微结构,从而修复UC大鼠结肠黏膜屏障功能。

Objective To assess the effect of optimized Kuijie Prescription on colon tissue in ulcerative colitis(UC)rats by regulating Toll-like receptor(TLR)/myeloid differentiation factor 88(MyD88)/nuclear factor kappa B(NF-κB).Methods A total of 40 male Sprague-Dawley(SD)rats were randomly divided into normal group,model group,optimized Kuijie Prescription group and sulfasalazine group,with 10 rats in each group.Except for the blank group,the other three groups were treated with trinitrobenzene sulfonic acid(TNBS)/ethanol secondary inflammation method combined with restraint method to establish UC rat model of qi stagnation and blood stasis type.After successful modeling,a 14-day intervention was conduted in Kuijie Prescription group(1.674 g/kg/d optimized Kuijie Prescription),sulfasalazine group(0.54 g/kg/d sulfasalazine),blank group and model group(water administration)to detect serum levels of interleukin-17A(IL-17A),IL-10,IL-22,and the protein levels of MyD88,TLR2,TLR4 and NF-κB p65 and the expression of mRNA in colon tissue of rats.Results Compared with blank group,the levels of IL-17A and IL-22 in model group were significantly increased(P<0.05),while the level of IL-10 was significantly decreased(P<0.05).Compared with model group,the serum levels of IL-17A and IL-22 in optimized Kuijie Prescription group and sulfasalazine group were significantly decreased(P<0.05),but the serum level of IL-10 was significantly increased(P<0.05).There was no significant difference in the serum levels of IL-17 A,IL-22 and IL-10 between optimized Kuijie Prescription group and sulfasalazine group(P>0.05).Compared with blank group,the protein levels of MyD88,TLR2,TLR4,NF-κB p65 and the expression of mRNA in colon tissue in model group were significantly increased(P<0.05).Compared with model group,the protein levels of MyD88,TLR2,TLR4,NF-κB p65 and the expression of mRNA in sulfasalazine group and optimized Kuijie Prescription group were significantly decreased(P<0.05).There was no significant difference in the protein levels of MyD88,TLR2,TLR4,NF-κB p65 and the expression of mRNA between optimized Kuijie Prescription group and sulfasalazine group(P>0.05).Conclusion Optimized Kuijie Prescription may reduce the serum level of inflammatory factors,improve the inflammatory response,and repair the ultrastructure of colon tissue by regulating the TLR/MyD88/NF-κB signaling pathway,thereby repairing the colonic mucus barrier function in UC rats.