Aim To investigate the change of coagulation and fibrinolysis functions of human microvascular endothelial cells (HMEC) induced by activated complement alternative pathway and effect of pyrrolidine-dithiocarbamate (PDTC) and resveratrol (Res) on intervention.Methods Normal human serum was activated by cobra venom factor (CVF).After exposure of HMEC to activated complement for various time points, supernatant was removed and assayed for activities of hydrolysing chromogenic substrate and affecting activated partial thromboplastin time (APTT) and prothrombin time (PT).The cells exposed to activated complement were collected and washed, and then the cell suspension was assayed for activity of affecting coagulation function of normal plasma.Lastly, the coagulation and fibrinolysis functions of HMEC pretreated with PDTC and Res were also investigated after HMEC was exposed to activated complement alternative pathway.Results The hydrolysis activity of chromogenic substrate of supernatant was up-regulated significantly after HMEC exposed to activated complement alternative pathway.The supernatant induced APTT decreased significantly, and also shortened PT.The cell suspension of various time points induced APTT decreased significantly, and also shortened PT by suspension of 6 h time point.PDTC and Res failed to inhibit the up-regulation of the chromogenic hydrolysis activity, but Res showed significant intervention on decrease of APTT, and PDTC had better effect on inhibiting the decrease of PT than that of Res.Conclusion Activated complement alternative pathway can induce abnormality of coagulation and fibrinolysis functions of HMEC, and PDTC and Res can affect this change.%目的 研究补体旁路激活导致的微血管内皮细胞纤溶凝血功能的变化以及吡咯烷二硫氨基甲酸(PDTC)和白藜芦醇(Res)的干预作用.方法 采用眼镜蛇毒因子激活血清补体旁路途径.将补体旁路活化的血清作用于人微血管内皮细胞(HMEC),检测多个时间点细胞培养上清的水解发色底物活性变化和对凝血酶原时间(PT)、活化部分凝血活酶时间(APTT)的影响,并制备细胞悬液,检测细胞表面的抗凝血功能变化情况.采用PDTC、Res对上述变化指标进行干预研究.结果 HMEC经补体旁路活化产物刺激后,细胞培养上清的发色底物酶解活性明显上调;细胞培养上清导致正常血浆的APTT明显缩短,同时,对PT也有缩短的作用;经补体旁路活化产物刺激HMEC后制备的各时间点细胞悬液对正常血浆的APTT有明显的缩短作用,6 h刺激组的细胞悬液也表现出缩短PT的作用.PDTC和Res未能抑制发色水解活性的上调,但Res对APTT的缩短有明显的干预作用,而PDTC能在一定程度上抑制PT的缩短.结论 补体旁路活化产物会导致HMEC纤溶凝血功能的失调,而PDTC和Res有一定的干预作用.
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