肾脏缺血再灌注损伤早期高迁移率族蛋白1对巨噬细胞和nTreg细胞功能的影响及其作用机制的实验研究

Renal ischemia and reperfusion injury(RIRI) is primary reason for high mortality rate of active kidney injury(AKI),however,the immune inflammator is one of the important mechanism of causing RIRI. Recently,HMGB1 was newly found a damage factor of inflammatory to promote strongly inflammatory cytokines that were secreted by macrophage and could decrease suppression function of Treg cell to result in and aggravate early injury on the organ of kidney in the initial stage of ischemia and reperfusion injury.At present,HMGB1 is regared as endogenous factor to induce and deteriotate the inflammatory in the eraly phase of RIRI.In this study, used C57BL/6 mice and TLR4 defective mice as test animal. separated and cultivated macrophages and Treg cells from spleen of mice and established RIRI animal model and used RT-PCR, western-blot,,FCM and ELISA ,cellular experiments in vitro,to observe early damage signal HMGB1 how to influence the function of the renal macrophage and nTreg cells .In accord to results,We analyze the affect and mechanism of HMGB1. This study will clarify the effect and significance of HMGB1 ,macrophage and Treg cells in initial stage of RIRIs. At the same time,we will explore to obstruct signal pathway of HMGB1/TLR4 which could produce protective effect of mice RIRI model and hope to find a new approach for development of drugs for this disease treatment.

肾脏缺血再灌注损伤（RIRI）是急性肾损伤高死亡率的主要原因，局部免疫炎症增强是引起RIRI的重要机制之一。肾脏缺血期坏死的肾上皮细胞可释放的高迁移率族蛋白（HMGB1)可促使再灌注早期巨噬细胞产生大量促炎性因子造成局部炎症增强，同时可降低Treg细胞对免疫炎症的抑制作用，加重早期损伤，是诱导早期炎症反应加强的关键内源性信号。本课题采用 C57BL/6 和同系TLR4缺陷小鼠制备RIRI实验模型，并在干预前后分离提取巨噬细胞和Treg细胞体外培养，以此动物模型和相关细胞为基础，采用ELISA、Western-blot、实时定量PCR、流式细胞术、磁珠分选及siRNA等方法分析HMGB1通过TLR4对RIRI早期肾脏巨噬细胞和Treg细胞功能影响的作用机制，局部炎症加强与肾损伤的关系，探讨阻断HMGB1/TLR4 信号通路对鼠RIRI的保护效应，为临床上更有效的防治RIRI提供新靶点和思路。

肾脏缺血再灌注损伤（RIRI）是急性肾损伤高死亡率的主要原因，局部免疫炎症增强是引起RIRI的重要机制之一。肾脏缺血期坏死的肾上皮细胞可释放的高迁移率族蛋白（HMGB1)可促使再灌注早期巨噬细胞产生大量促炎性因子造成局部炎症增强，同时可降低Treg细胞对免疫炎症的抑制作用，加重早期损伤，是诱导早期炎症反应加强的关键内源性信号。本课题采用 C57BL/6 和同系TLR4缺陷小鼠制备RIRI实验模型，并在干预前后分离提取巨噬细胞和Treg细胞体外培养，以此动物模型和相关细胞为基础，采用ELISA、Western-blot、实时定量PCR、流式细胞术、磁珠分选及siRNA等方法分析HMGB1通过TLR4对RIRI早期肾脏巨噬细胞和Treg细胞功能影响的作用机制，局部炎症加强与肾损伤的关系，探讨阻断HMGB1/TLR4 信号通路对鼠RIRI的保护效应，为临床上更有效的防治RIRI提供新靶点和思路。