中文摘要：

目的观察高迁移率族蛋白B1（hiigh mobility groupp rotein box1，HMGB1）介导肾小管上皮细胞功能的变化并探讨其机制。方法体外培养大鼠肾小管上皮细胞（NRK52E），分为空白对照组、HMGB1刺激组以及HMGB1＋红假单胞菌脂多糖（Lipopolysaccharide from Rhodobactersphaeroides，LPSRS）刺激组，采用免疫荧光及Western印迹检测Toll样受体4（Toll-likereceptor4，TLR4）表达，流式细胞术检测细胞凋亡率及细胞周期变化，Westem印迹检测MAPK信号通路及NF—KB激活情况，实时荧光定量PCR检测细胞IL-1、IL-6及组织金属蛋白酶2抑制因子（tissue inhibitor of metalloprUteinases2，TIMP2）mRNA表达水平，蛋白芯片检测IL-1、IL-6及TIMP2蛋白表达水平。结果NRK52E细胞表达TLR4。与对照组比较，HMGB1刺激组细胞周期G1期阻滞率高，MAPK信号通路及NF-kB激活，IL-1、IL-6及TIMP2 mRNA及蛋白表达水平均显著升高（均P〈0．05）。TLR4特异性阻断剂LPSRS可以显著抑制HMGB1所介导的效应（均P〈0．05）。结论 HMGB1与NRK52E细胞上TLR4的相互作用介导肾小管上皮细胞炎性反应，主动合成并释放炎性介质。

英文摘要：

Objective To observe functional changes of renal tubular epithelial cells stimulated by high mobility group protein box 1 （HMGB1） and associated mechanism. Methods Renal tubular epithelial cells （NRK52E） were divided into control group, HMGB1 group and HMGBI＋ lipopolysaccharide from Rhodobaetersphaeroides （LPS RS） group. Toll-like receptor 4 （TLR4） expression was detected by immunofluorescence and Western blotting. Apoptosis rate and cell cycle arrest were identified with flow eytometry. The activation of MAPK signaling pathway and NF-KB were detected by Western blotting. The IL- 1, IL- 6 and tissue inhibitor of metalloproteinases 2 （TIMP2） mRNA levels were measured by real- time PCR. The secretion levels of IL- 1, IL- 6 and TIMP2 were measured by protein chips assay. Results TLR4 was expressed by NRK52E ceils. Compared with the control group, there were increased cell cycle G1 arrest, MAPK signaling pathway and NF-KB activation in HMGB1 group. Furthermore, IL-1, IL-6 and TIMP2 mRNA levels were increased and IL-1, IL-6 and TIMP2 were secreted by NRK52E when stimulated with HMGB1 （all P〈0.05）. However, effects mediated by HMGB1 stimulation could be inhibited by LPS RS （all P〈0.05）. Conclusions Inflammatory activation of NRK52E cells can be mediated by the interaction of HMGB1 and TLR4.