کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
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6002349 | 1182967 | 2014 | 8 صفحه PDF | دانلود رایگان |
![عکس صفحه اول مقاله: The neutrophil serine protease PR3 induces shape change of platelets via the Rho/Rho kinase and Ca2 + signaling pathways The neutrophil serine protease PR3 induces shape change of platelets via the Rho/Rho kinase and Ca2 + signaling pathways](/preview/png/6002349.png)
IntroductionProteinase 3 (PR3) is released from neutrophil azurophilic granules and exerts complex effects on the inflammatory process. PR3 catalyzes the degradation of a number of macromolecules, but the consequences on blood cells are less well defined. In the present study, the effect of PR3 on human platelets was thoroughly investigated.MethodsThe experiments were performed on washed platelets freshly isolated from blood donated by healthy human volunteers. Platelets shape change and aggregation was measured on a Chrono-Log aggregometer. The phosphorylated form of MYPT1 was visualized by immunostaining. Platelet activation was further evaluated by flow cytometry.ResultsPR3 induced platelet shape change but not aggregation. Flow cytometry analysis showed that PR3 induced no P-selectin expression or binding of fibrinogen to the platelets, and it did not change the activation in response to PAR1- or PAR4-activating peptides or to thrombin. Furthermore, Fura-2 measurement and immuno-blotting analysis, respectively, revealed that PR3 stimulated small intracellular Ca2Â + mobilization and Thr696-specific phosphorylation of the myosin phosphatase target subunit 1 (MYPT1). Separate treatment of platelets with the Rho/Rho kinase inhibitor Y-27632 and the intracellular Ca2Â + chelator BAPTA/AM reduced the shape change induced by PR3 whereas concurrent treatment completely inhibited it.ConclusionThe data shows that the neutrophil protease PR3 is a direct modulator of human platelets and causes shape change through activation of the Rho/Rho kinase and Ca2Â + signaling pathways. This finding highlights an additional mechanism in the complex interplay between neutrophils and platelets.
Journal: Thrombosis Research - Volume 134, Issue 2, August 2014, Pages 418-425