补体旁路激活致内皮细胞纤溶凝血相关分子表达变化及干预研究

摘要

Aim To investigate the change of molecu-lar expression related to coagulation and fibrinolysis in human microvascular endothelial cells ( HMEC ) in-duced by activated complement alternative pathway and effect of pyrrolidine dithiocarbamate ( PDTC ) and res-veratrol on intervention. Methods Normal human se-rum was activated by cobra venom factor ( CVF) . After exposure of HMEC to activated complement for various times, supernatant was removed and assayed for ex-pressions of P-selectin, VWF, t-PA, PAI-1, TF, TM, and NO by using reagent kits. The expressions of the above molecules in HMEC pretreated with PDTC and resveratrol were also investigated. Results P-selectin and VWF were rapidly released by endothelial cells and the expression reached the peak at the time point of 15 min. The expressions of t-PA, PAI-1, and TF were continuously upregulated, whereas NO and TM were decreased. PDTC and resveratrol inhibited the upregulation of P-selectin, VWF, t-PA, PAI-1 and TF, and intervened the downregulation of NO. Res-veratrol further downregulated the expression of TM. Conclusion Activated complement alternative path-way can influence the expression of molecules related to coagulation and fibrinolysis in HMEC, and PDTC and resveratrol can affect this change.%目的：研究补体旁路激活致微血管内皮细胞纤溶凝血相关分子表达的变化及干预。方法采用眼镜蛇毒因子激活血清补体旁路途径。将旁路活化的血清作用于人微血管内皮细胞,通过ELISA检测多个时间点细胞培养上清中P-selectin、VWF、t-PA、PAI-1、TF、TM 的表达,采用 NO 试剂盒检测NO水平,并进一步研究吡咯烷二硫氨基甲酸( PDTC)、白藜芦醇对以上指标变化的影响。结果微血管内皮细胞经补体旁路活化产物刺激后,P-selectin、VWF的表达快速上调,其高峰时间为15 min,而纤溶功能分子t-PA、PAI-1也随后出现持续上调表达,与凝血功能相关的组织因子TF的表达水平持续上调,而血栓蛋白 TM 及 NO 的表达下降。PDTC、白藜芦醇对P-selectin、VWF、t-PA、PAI-1、TF上调表达具有抑制作用,对NO下调表达具有干预作用。而白藜芦醇能使TM的表达进一步下调。结论补体旁路活化产物会导致微血管内皮细胞纤溶凝血功能相关分子表达的变化,而PDTC、白藜芦醇对此种变化具有一定的影响。