摘要
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.
Abstract
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 Pre-scription 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 second-ary inflammation method combined with restraint method to estab-lish UC rat model of qi stagnation and blood stasis type.After successful modeling,a 14-day intervention was conduted in Kui-jie 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 Com-pared 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 pro-tein 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 expres-sion 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 opti-mized 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.
基金项目
陕西省中医药管理局秦创原中医药创新研发转化项目(2022-QCYZH-015)
陕西省科学技术厅陕西自然科学基础研究计划项目(2022JM-489)
内蒙古自治区卫生健康委员会2022年度自治区卫生健康科技计划项目(202201599)
内蒙古自治区科技计划项目(2022YFSH0003)
2023年度河北省社会科学发展研究课题(20230204037)
国家科技期刊平台
NSTL
万方数据