title: [PI3-kinase mediates activity of RhoA and interaction of RhoA with mDia1 in thrombin-induced platelet aggregation].
abstract: The aim of this study was to investigate the role of RhoA/mDia1 pathway in the process of thrombin-induced platelet aggregation and regulatory effect of PI3K inhibitor on this process. The human platelets were isolated from peripheral blood, the activation of RhoA, Rac1 and Cdc42 in the platelet aggregation was detected by GST pull-down assay and immune co-precipitation, the interaction of RhoA, Rac1 and Cdc42 with mDia1 and the formation of complex in the process of platelet aggregation were determined by immune coprecipitation, and the effect of PI3K inhibitor (wortmannin) on above-mentioned process was assayed. The results showed that thrombin elevated the activity of RhoA and the binding capability of RhoA with mDia1 during thrombin-induced platelet aggregation and spreading on Fg coated coverslips. Wortmannin inhibited the rising of RhoA activity and the binding level of RhoA with mDia1 induced by thrombin. Thrombin elevated the activity of Rac1 and Cdc42 during thrombin-induced platelet aggregation, but could not induce binding of Rac1 or Cdc42 with mDia1. Wortmannin could not inhibit the rising of Rac1 and Cdc42 activity induced by thrombin. It is concluded that the PI3-kinase regulates the thrombin-induced actin cytoskeleton reconstitution in platelets by RhoA-mDia1 pathway.
authors.affiliation: Department of Hematology, Xijing Hospital, Fourth Military Medical University, Xi'an, 710032, Shaanxi Province, China.
authors.forename: Guang-Xun
authors.initials: GX
authors.lastname: Gao
authors.forename: Hong-Juan
authors.initials: HJ
authors.lastname: Dong
authors.forename: Hong-Tao
authors.initials: HT
authors.lastname: Gu
authors.forename: Ying
authors.initials: Y
authors.lastname: Gao
authors.forename: Yao-Zhu
authors.initials: YZ
authors.lastname: Pan
authors.forename: Yang
authors.initials: Y
authors.lastname: Yang
authors.forename: Xie-Qun
authors.initials: XQ
authors.lastname: Chen
mesh_terms: D000199:Actins / Q000378:metabolism / Q000494:pharmacology; D048868:Adaptor Proteins, Signal Transducing / Q000276:immunology / Q000378:metabolism*; D001792:Blood Platelets / Q000378:metabolism*; D002478:Cells, Cultured; D000080865:Formins; D006801:Humans; D019869:Phosphatidylinositol 3-Kinases / Q000494:pharmacology*; D010974:Platelet Aggregation / Q000187:drug effects*; D013917:Thrombin / Q000494:pharmacology; D020830:rac1 GTP-Binding Protein / Q000378:metabolism; D020742:rhoA GTP-Binding Protein / Q000378:metabolism* / Q000494:pharmacology
chemical_list: D000199:Actins; D048868:Adaptor Proteins, Signal Transducing; D000080865:Formins; D019869:Phosphatidylinositol 3-Kinases; D013917:Thrombin; D020830:rac1 GTP-Binding Protein; D020742:rhoA GTP-Binding Protein; C108962:DIAPH1 protein, human