TLR4在LPS诱导的结肠炎症恢复中的作用

发布时间：2018-04-20 00:16

本文选题：炎症性肠病 + Toll样受体　；参考：《中国病理生理杂志》2017年02期

【摘要】：目的:研究Toll样受体4(Toll-like receptor 4,TLR-4)在结肠炎症中的作用,探讨LPS在炎症性肠病中的治疗作用。方法:取正常肠上皮细胞进行体外脂多糖(lipopolysaccharide,LPS)干预培养。采用慢病毒转染技术,构建TLR4低表达、正常表达及高表达的肠上皮细胞亚组。正常表达组(normal组)及高表达组(high组)培养基中加入LPS诱导细胞炎症,刺激时间分别为0、2、4 h。Western blot法检测TLR4的表达;收集细胞上清液,ELISA检测各亚组细胞炎症因子TNF-α、IL-6和IL-8的水平。收集细胞,qPCR检测细胞因子TNF-α、IL-6、IL-8、IL-10和IL-1βmRNA的表达水平。划痕试验观察对比2组细胞的迁移能力。结果:LPS干预培养细胞后,TLR4的表达量显著增加(P0.05)。ELISA和qPCR检测高表达组与正常表达组组间细胞因子TNF-α、IL-6、IL-8、IL-10和IL-1β的蛋白和mRNA水平的差异均有统计学意义(P0.05)。划痕试验提示TLR4高表达组的细胞迁移能力明显高于正常对照组。结论:LPS影响TLR4炎症通路的活化,促进前炎症因子及辅助刺激分子的释放,起到调节炎症反应的作用。
[Abstract]:Objective: to study the role of Toll like receptor 4(Toll-like receptor 4 (TLR 4) in colitis and to explore the therapeutic effect of LPS in inflammatory bowel disease. Methods: normal intestinal epithelial cells were cultured with lipopolysaccharide in vitro. Lentivirus transfection technique was used to construct a subgroup of intestinal epithelial cells with low expression, normal expression and high expression of TLR4. Normal expression group (normal group) and high expression group (high group) were added to the culture medium to induce cell inflammation, the stimulation time was 0 ~ 2G ~ 4 h.Western blot method to detect the expression of TLR4, and the levels of TNF- 伪 IL-6 and IL-8 in each subgroup were detected by Elisa. The expression levels of IL-10 and IL-1 尾 mRNA were detected by qPCR. The migration ability of the two groups was observed by scratch test. Results the expression of TLR4 in cultured cells was significantly increased after the intervention of TNF- 伪, IL-6, IL-8, IL-10 and IL-1 尾, and the levels of mRNA were significantly different between the high expression group and the normal expression group by Elisa and qPCR. Scratch test showed that the cell migration ability of TLR4 high expression group was significantly higher than that of normal control group. ConclusionLipopolysaccharide affects the activation of TLR4 inflammatory pathway, promotes the release of proinflammatory factors and costimulatory molecules, and regulates the inflammatory response.
【作者单位】：广州医科大学附属广州市第一人民医院胃肠外科;
【基金】：广州市科技计划(No.2014Y2-00074) 广东省自然科学基金资助项目(No.2015A030313729)
【分类号】：R574.62

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