蛋白激酶C抑制剂Staurosporine对血小板聚集、蛋白磷酸化和肌动蛋白聚合的影响  

EFFECTS OF AN INHIBITOR OF PROTEIN KINASE C, STAUROSPORINE,ON PLATELET AGGREGATION,PROTEIN PHOSPHORYLATION AND ACTIN POLYMERIZATION

作　　者：陈日炎[1] 覃燕梅[1] 江黎明[1] 梁念慈[1] 

机构地区：[1]广东医学院生化教研室,湛江524023

出　　处：《广东医学院学报》1997年第4期309-312,共4页Journal of Guangdong Medical College

基　　金：国家自然科学基金

摘　　要：目的：进一步研究蛋白激酶C抑制剂Staurosporine在血小板聚集、蛋白磷酸化、肌动蛋白聚合中的作用。方法：以32P－Na2HPO4标记血小板；以血小板聚集仪测定血小板聚集；以SDS－PAGE分离蛋白质，进行放射自显影；以TritonX－100抽提法沉淀骨架蛋白。结果：（1）1μmol／LStaurosporine完全抑制0．5U／ml凝血酶或10μmol／LPMA诱导的血小板聚集，大部分抑制20μmol／LA23187诱导的血小板聚集。（2）1μmol／LStaurosporine几乎完全抑制0．5U／ml凝血酶、或10μmol／LPMA、或20μmol／LA23187诱导的血小板40kD和20kD蛋白磷酸化。（3）1μmol／LStaurosporine完全抑制0．5U／ml凝血酶或10μmol／LPMA诱导的血小板肌动蛋白聚合。结论：Staurosporine抑制蛋白激酶C的活性．从而抑制血小板的聚集和肌动蛋白聚合；蛋白激酶C在血小板聚集和肌动蛋白聚合中起着重要调控作用。Objective: To study further the role of staurosporine, an inhibitor of protein kinase C, on platelet aggregation, protein phosphorylation and actin polymerization. Methods: Platelet was labelled with32P-Na2HPO4;platelet aggregation was rneasured by aggregometer;proteins were separated by SDS-PAGE and phosphorylated proteins were detected by autoradiophotograph; cytoskeletal proteins were precipitated by Triton X-100 extraction buffer. Results: (1) 1μmol/L staurosporine completely inhibited 0. 5U/ml thrombin or 10μmol/L PMA-induced platelet aggregation, and largely inhibited 20 μmol/L A23187-induced platelet aggregation, respectively; (2) 1μmol/L staurosporine completely inhibited 0. 5 U/ml thrombin,or 10μmol/L PMA or 20μmol/L A23187-induced the phosphorylation of two proteins，a 40kD substrate of protein kinase C (pleckstrin ), and the 20kD myosin light chain (MLC); and (3)1μmol/L staurosporine completely suppressed actin polymerization enhanced by 0, 5U/L thrombin or 10μmol/L PMA platelet. Conclusions: Staurosporine can inhibit platelet aggregation and actin polymer-ization through inhibiting protein kinase C;protein kinase C may play an important role in platelet ag-gregation and actin polymerization.

关 键 词：蛋白激酶C 抑制剂 血小板聚集 肌动蛋白聚合 

分 类 号：Q592.1[生物学—生物化学] R973.2[医药卫生—药品]