Toll样受体介导的NF-κB信号通路在抗结核药物致肝损伤中的作用及狗肝菜多糖（DCP）干预机制研究

In the process of TB treatment, the most frequent and serious adverse reactions of the antituberculosis drugs-induced liver injury. For research the role of Toll-like Receptors TLR2 and TLR4 mediated NF-κB Signaling pathway in Anti-tuberculosis drugs induced Liver Injury , to built the antituberculosis drugs-induced liver injury in culture primary hepatocyte model, Detection the expression of reaction related parameters of liver injury and TLR2 , TLR4, MyD88, NF kappa B. To use TLR2 and TLR4 siRNA and slow virus up and down regulation the TLR2 and TLR4 expression respectively, observe the role of TLR2 and TLR4 in the NF-kappa B. With MyD88 and nf-kappa B blockers to observe whether TLR2 and TLR4 mediated the NF-kappa B signaling pathways during ATLI. Animals and TLR2 and TLR4, the NF-kappa B gene knockout mice to investigate TLR2 and TLR4 mediated the NF-kappaB mechanism in ATLI. Combining the above experiments, and at the same time use DCP to intervention.To show whether in the process of ATLI , the protective role DCP by TLR2 and TLR4 mediated the NF-kappa B signaling pathways,The experiment may provide a new therapeutic targets for the prevention the ATLI and potential treatment drugs.

治疗结核病过程中频发且严重不良反应是抗结核药物所致肝损伤（ATLI）。为探讨Toll样受体TLR2和TLR4介导NF-κB通路在ATLI中的作用，建立抗结核药致小鼠原代肝细胞损伤模型，检测反应肝损伤相关指标及TLR2 、TLR4、MyD88、NF-κB的蛋白表达；利用针对TLR2、TLR4的小干扰RNA和慢病毒表达载体分别上、下调TLR2、TLR4观察TLR2、TLR4对NF-κB的作用；分别用MyD88和NF-κB的阻断剂来观察TLR2、TLR4是否介导NF-κB通路对ATLI产生作用。进一步应用TLR2、TLR4、NF-κB基因敲除小鼠佐证TLR2、TLR4介导NF-κB通路在ATLI中的机理。结合以上实验，同时给予狗肝菜多糖DCP干预，探讨在ATLI过程中DCP是否通过TLR2、TLR4介导NF-κB信号通路发挥保护作用。本项目的研究将可能为防治ATLI提供新的治疗靶点和潜在药物。

在治疗结核病过程中发生频率较高且较严重的不良反应是抗结核药物所致肝损伤（ATLI），课题组通过本研究建立了抗结核药物（INH+RFP）致小鼠原代肝细胞损伤模型以及抗结核药物致野生型和TLR4-/-基因敲除小鼠肝损伤动物模型，研究了TLR2、TLR4、MyD88、NF-κB 在ATLI中的相互调控关系；明确TLR4可通过调节MyD88和NF-κB对ATLI产生作用 ；证实了狗肝菜多糖（DCP）对ATLI具有良好保护作用其机理主要通过TLR4介导NF-κB信号通路起调节保护作用。此外，利用TLR4-/-小鼠研究了狗肝菜多糖可通过TLR4/ NF-κB信号通路对肝脏代谢紊乱起改善调节作用和治疗免疫性肝损伤的研究；以及基于TLR4/ NF-κB信号通路开展了相关药物对肝损伤防治作用的研究工作。为狗肝菜多糖（DCP）在肝损伤治疗的临床应用提供了更多实验及理论依据。