活化血小板对巨噬细胞分泌尿激酶型纤溶酶原激活物的影响  

Effect of activated platelets on the expression of urokinase-type plasminogen activator secreted by macrophages

作　　者：邹继宏[1,2] 杨丽霞[1] 郭瑞威[1] 刘宏[1] 朱国富[1] 梁星[1] 刘高[1,2] 

机构地区：[1]成都军区昆明总医院,昆明650032 [2]昆明医科大学成都军区昆明总医院临床学院,昆明650032

出　　处：《岭南心血管病杂志》2013年第5期616-620,625,共6页South China Journal of Cardiovascular Diseases

基　　金：成都军区医学科研"十一五"计划课题(项目编号:MB09029)

摘　　要：目的探讨活化血小板对巨噬细胞分泌尿激酶型纤溶酶原激活物(urokinase-type plasminogen activator,uPA)的影响。方法分离昆明小鼠血小板并计数,使用二磷酸腺苷(adenosine diphosphate,ADP)对血小板进行活化,采用流式细胞学技术检测血小板活化程度;分别在12 h、15 h、18 h和21 h时用实时聚合酶链反应(realtime polymerase chain reaction,rtPCR)和酶联免疫吸附测定(enzyme linked immunosorbent assay,ELISA)法检测活化血小板刺激小鼠巨噬细胞Raw264.7中uPA表达的影响。再采用活化和未活化血小板刺激小鼠巨噬细胞,检测uPA的表达。结果成功分离鉴定血小板。流式细胞学技术检测结果显示,10、20、40μmol/L的ADP诱导血小板表面的P选择素表达比0μmol/L时的ADP高,但差异无统计学意义(P>0.05)。20μmol/L的ADP活化血小板后刺激巨噬细胞18 h组的血小板P选择素表达比12 h、15 h、21 h组明显升高,差异有统计学意义(P<0.05)。分别用0、20μmol/L ADP活化血小板后刺激巨噬细胞18 h,结果显示20μmol/L ADP组比0μmol/L ADP组血小板P选择素表达明显升高,差异有统计学意义(P<0.05)。结论活化血小板能诱导巨噬细胞uPA的表达,从而可能在动脉粥样硬化斑块的稳定性中发挥作用。Objectives To determine the effect of activated platelets on the expression of urokinase-type plasminogen activator （uPA） secreted by macrophages.Methods Different concentrations of adenosine diphosphate （ADP） were used to stimulate platelets and the mean fluorescence of CD62PE was analyzed by Flow cytometry.Expressions of uPA at different times （12 h,15 h,18 h and 21 h） were analyzed by real-time polymerase chain reaction （rtPCR） and enzyme-linked immunosorbent assay （ELISA）.Activated and unactivated platelets were cocultured with macrophages for 18 h before rtPCR and ELISA were used to analyze the expression of uPA.Results Platelets were successfully separated and identified.With the concentrations of 10,20,40 μmol/L ADP,mean fluorescences of CD62PE were higher than that of unstimulated platelets,but there was no significant difference among them （P〉0.05）.When platelets were stimulated by 20 μmol/L ADP for 18 h,expression of uPA secreted by macrophages was obviously the highest compared with those at 12 h,15 h and 21 h P〈0.05.Compared with the unactivated platelets group,expression of uPA secreted by macrophages when platelets were stimulated by 20 μmol/L ADP was significantly higher （P〈0.05）.Conclusions Activated platelets can stimulate the expression of uPA secreted by macrophages,thus,they may affect the stableness of atherosclerotic plaque.

关 键 词：冠状动脉疾病 活化血小板 巨噬细胞 尿激酶型纤溶酶原激活物 不稳定斑块 

分 类 号：R541.4[医药卫生—心血管疾病]