活化血小板参与缺血性脑卒中炎症反应机制初探

Involvement of activated platelet in inflammatory response in ischemic stroke

目的 通过细胞共培养模型初步探讨活化血小板参与缺血性脑卒中炎症反应的机制.方法 分离健康人血小板,以0.8μmol/L二磷酸腺苷（ADP）作用20 min激活.同时设静息血小板做为对照,应用血细胞分析仪检测血小板平均内含物（MAC）浓度;将静息血小板和活化血小板分别与人脐静脉内皮细胞株共培养12 h,同时设ADP对照组（只加入等量ADP,无血小板）和空白对照组,采用RT-PCR法检测人脐静脉内皮细胞炎性细胞因子白介素-1β（IL-1β）、白介素.6（IL-6）、白介素-8（IL-8）、单核细胞趋化蛋白-1（MCP-1）的mRNA表达情况.结果与静息血小板组[（266.80±22.14）g/L]比较,ADP诱导激活的活化血小板MPC值[（231.90±14.63）g/L]减低,差异有统计学意义（P〈0.05）;RT-PCR检测结果显示活化血小板可以诱导共培养的人脐静脉内皮细胞表达IL-1β、IL-6、IL-8、MCP-1 mRNA,而静息血小板组、ADP对照组和空白对照组均无上述细胞因子的表达.结论 体外活化的血小板能够启动血小板一内皮细胞反应,诱导其表达炎症细胞因子,是缺血性脑卒中炎症反应发生的可能机制之一.

Objective To preliminarily study the role of activated platelets in inflammatory response in ischemic stroke by using the platelet and vascular endothelial cell co-culture model.Methods Platelets were isolated from healthy donator＇s peripheral blood and activated in vitro by 0.8 μmol/L ADP in ischemic stroke; resting platelets were also chosen.The mean platelet component （MPC）concentration was measured by hematology analyzer.Resting platelets and activated platelets were co-cultured with vascular endothelial cells for 12 h,respectively; the other 2 control groups （ADP only,blank） were employed in the experiment.RT-PCR was used to detect the expression level of mRNA of proinflammatory cytokines in the endothelial cells,including IL-1β,IL-6,IL-8 and MCP-1.Results Compared with the resting platelets[（266.80±22.14） g/L],ADP-induced activated platelets[（231.90±14.63） g/L]showed significantly lower concentration of MPC （P〈0.05）.Activated platelets in the co-culture model could induce the mRNA expression of IL-1β,IL-6,IL-8 and MCP-1 in the endothelial cells,while resting platelets could not.Conclusion Activated platelets can trigger an interaction between platelets and endothelial cells in vitro,and induce the mRNA expression of proinflammatory cytokines in the endothelial cells,which may be one of the mechanisms of inflammatory response in iscbemic stroke.