NF-κB在炎症介质诱导的大鼠脑微血栓形成分子发病机制中的作用△

The Role of Nuclear Factor-Kappa B in Molecular Mechanism of Pathogenesis of Rat Brain Microvascular Thrombosis Induced by Inflammatory Mediators

目的以脂多糖(LPS)诱导下高表达组织因子(tissue factor,TF)的大鼠脑微血管内皮细胞(BMECs)为大鼠脑微血栓形成的细胞模型,探讨核因子κB(NF-κB)在炎症介质导致的脑微血栓形成中的作用。方法制备大鼠脑微血管内皮细胞培养模型,分别用LPS刺激1/4、1/2、1、2、4、16 h,以NF-κB特异性抑制剂PDTC(pyrrilidine dithiocarbam-ate)预处理组和空白对照组作为对照,采用流式细胞术特异性检测经BMECs膜表面TF定位表达丰度的变化,同时检测胞浆中TF的mRNA水平的变化,并进行计算机定量分析。结果TF在BMECs细胞中的表达与LPS作用呈现时间依赖关系,10μg/ml LPS刺激1/2 h时TF表达开始升高,1 h时达到最高值;100μmol/L PDTC预处理的BMECs在LPS刺激下,TF表达水平不升高。结论LPS可以显著诱导BMECs中TF的表达,NF-κB是LPS诱导的BMECs TF表达的关键性转录调控分子,是炎症介质诱导的脑血栓形成发病分子机制中的重要因素。

Objective To investigate the role of nuclear factor-kappa B （NF-KB） in tissue factor（TF） induction by inflammatory mediator lipopolysaccharide （LPS） in cultured rat brain microvascular endothelial cells（BMECs）. Methods The BMECs were isolated and cuhured, Pyrrilidine dithiocarbamate （PDTC）,a special inhibitor of NF- KB pathway was used to preincubated on BMECs, TF expression of BMECs in response to LPS was examined by flow cytometry and RT-PCR, Results TF expression showed a time-dependent manner of LPS, TF expression of BMECs began to rise after induced with LPS for 1/2 h, reached top when induced for 1 h ;TF expression did not rise when BMECs were preincubated by 100 μmol/L PDTC. Conclusion NF-kB play a critical role in regulating TF transcription in LPS-induced BMECs and is a core factor in molecular mechanism of pathogenesis of cerebral thrombosis induced by inflammatory mediators.