白藜芦醇对溃疡性结肠炎小鼠肠上皮Toll样受体4/髓样分化因子/核转录因子-κB信号通路的影响

Effect of resveratrol on colonic epithelium Toll-like receptor 4/myeloid differentiation factor 88/nuclear transcription factor-kappa B p65 signaling pathway in mice with ulcerative colitis

目的探讨白藜芦醇(RES)对葡聚糖硫酸钠盐(DSS)诱导溃疡性结肠炎(UC)小鼠的治疗效果,及其对Toll样受体4(TLR4)/髓样分化因子88(MYD88)/核转录因子-κB(NF-κB)信号通路的调控机制。方法将90只BALB/c小鼠的体重通过SPSS生成随机数后,参照其大小排序分为对照(CON)组、模型(DSS)组、白藜芦醇低(RES-L,10 mg/kg)、中(RES-M,50 mg/kg)、高(RES-H,100 mg/kg)剂量组、柳氮磺砒啶(SASP)组。记录小鼠状况,测算疾病活动指数(DAI),实时荧光定量聚合酶链式反应(RT-PCR)和蛋白免疫印迹法(Western blot)检测结肠上皮组织中TLR4、MYD88、NF-κB p65 mRNA和蛋白的表达;酶联免疫吸附法(ELISA)检测小鼠结肠肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)、白细胞介素-1β(IL-1β)、白细胞介素-10(IL-10)的含量,并用Graph Pad Prism 8统计软件分析。结果DSS组TLR4、MYD88、NF-κB p65显著高于CON组(0.315±0.046比0.082±0.011、0.279±0.025比0.085±0.012、0.244±0.026比0.052±0.011,t=8.352、12.045、11.476,P<0.05),差异有统计学意义,RES-H的TLR4、MYD88、NF-κB p65显著低于DSS组(0.105±0.016比0.311±0.045、0.094±0.012比0.295±0.021、0.103±0.015比0.245±0.026,t=9.174、7.285、11.163,P<0.05),差异有统计学意义;RES-H组TNF-α、IL-1β、IL-6显著低于DSS组(154.105±6.184比303.408±31.209、93.522±7.271比306.305±40.926、140.509±2.795比263.705±11.116,t=8.174、8.865、18.622,P<0.05),差异有统计学意义,RES-H组IL-10高于DSS组(110.708±4.154比93.846±3.232,t=5.565,P<0.05),差异有统计学意义。结论RES通过调节TLR4/MYD88/NF-κB p65信号通路活化达到治疗UC的目的。

Objective To study the therapeutic efficacy of resveratrol(RES)for dextran sodium sulfate(DSS)-induced ulcerative colitis(UC)in mice and its regulatory effect on Toll-like receptor 4(TLR4)/myeloid differentiation factor 88(MyD88)/nuclear factor-κB(NF-κB)signaling pathway.Methods After the weights of 90 BALB/c mice were randomly generated by SPSS,the mice were sorted according to their size and divided into control(CON)group and model(DSS)group according to their body weight,with low resveratrol(RES-L,10 mg/kg),medium(RES-M,50 mg/kg),high(RES-H,100 mg/kg)dose group,sulfasalazine(SASP)group.Status of mice was recorded,and disease activity index(DAI)was measured.The real-time fluorescent quantitative polymerase chain reaction(Real-time PCR)and Western blotting used to detect the expression of TLR4,MYD88 and NF-κB p65 mRNA and protein in colonic epithelial tissues respectively.The enzyme-linked immunosorbent assay(ELISA)was used to detect colonic tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),IL-1βand IL-10 contents.GraphPad Prism 8 statistical software was used.Results The expression of TLR4,MYD88,and NF-κB p65 in the DSS group was significantly higher than that in the CON group(0.315±0.046 vs.0.082±0.011,0.279±0.025 vs.0.085±0.012,0.244±0.026 vs.0.052±0.011,t=8.352,t=12.045,t=11.476,P<0.05).The expression of TLR4,MYD88 and NF-κB p65 in the RES-H group was significantly lower than in the DSS group(0.105±0.016 vs.0.311±0.045,0.094±0.012 vs.0.295±0.021,0.103±0.015 vs.0.245±0.026,t=7.285,t=11.163,t=9.174,P<0.05).TNF-α,IL-1βand IL-6 levels in the RES-H group were significantly lower than those in the DSS group(154.105±6.184 vs.303.408±31.209,93.522±7.271 vs.306.305±40.926,140.509±2.795 vs.263.705±11.116,t=8.174,t=8.865,t=18.622,P<0.05).The IL-10 level in the RES-H group was significantly higher than that in the DSS group(110.708±4.154 vs.93.846±3.232,t=5.565,P<0.05).Conclusion RES can treat UC by regulating the activation of TLR4/MYD88/NF-κB p65 signaling pathway.