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目的探讨Toll样受体4(TLR4)单克隆抗体(TLR4mAb)对葡聚糖硫酸钠(DSS)诱导的急性期溃疡性结肠炎(UC)小鼠肠黏膜TLR4介导的核因子(NF)-κB信号通路中磷酸化IκB激酶(p-IKK)及NF-κB的影响情况。方法30只BALB/c小鼠均分为正常对照组(A组)、UC模型组(B组)及低、中、高剂量TLR4mAb干预组(C、D、E组)。A组小鼠饮用蒸馏水7d;B、C、D、E组小鼠饮用5%DSS水溶液7d以制成UC模型。造模同时,C、D、E组小鼠分别腹腔注射低、中、高剂量TLR4mAb。造模及干预7d后处死小鼠,观察指标包括疾病活动指数(DAI)、结肠组织病理学评分(HPS)。采用Western印迹法检测各组肠黏膜p-IKK的蛋白表达,蛋白凝胶电泳迁移率变动分析法(EMSA法)检测NF-κB的活性变化。结果B组小鼠的结肠黏膜DAI及HPS均显著高于A组(P值均<0.01)。与模型组相比,使用TLR4mAb干预后中、高剂量组HPS有不同程度的缓解(P值均<0.01)。②与B组相比,D、E组的TLR4mAb后p-IKK表达及NF-κB的活性均显著下降(P值分别<0.05、0.01)。结论TLR4mAb可以减轻肠道炎症,发挥干预作用,其机制可能是通过抑制TLR4介导的NF-κB通路关键蛋白的表达,降低NF-κB的活性,继而减少下游炎性因子过度表达。
Objective To investigate the effects of Toll - like receptor 4 (TLR4) monoclonal antibody (TLR4 mAb) on TLR4 - mediated nuclear factor (NF) - κB signaling pathway phosphorylated IκB kinase (p-IKK) and NF-κB impact. Methods Thirty BALB / c mice were divided into normal control group (A group), UC model group (B group) and low, medium and high doses of TLR4 mAb intervention group (C, D and E). Group A mice drank distilled water for 7 days; Group B, C, D and E mice drank 5% DSS aqueous solution for 7 days to make UC model. At the same time, C, D, E mice were injected intraperitoneally with low, medium and high doses of TLR4 mAb respectively. Mice were sacrificed 7 days after modeling and intervention. The indexes included disease activity index (DAI) and colon histopathology score (HPS). The protein expression of p-IKK in intestinal mucosa of each group was detected by Western blotting, and the change of NF-κB activity was detected by EMSA. Results The colonic mucosa DAI and HPS in group B were significantly higher than those in group A (all P <0.01). Compared with the model group, HPS was moderately and moderately relieved after TLR4mAb intervention (P <0.01). ② Compared with group B, the expression of p-IKK and the activity of NF-κB in group D and E decreased significantly after TLR4 mAb (P <0.05, 0.01 respectively). Conclusion TLR4 mAb can relieve intestinal inflammation and play an intervention role. The possible mechanism is that TLR4 can reduce the expression of NF-κB pathway-mediated key proteins, decrease the activity of NF-κB and then decrease the expression of downstream inflammatory factors.