[Activation of Hippo Pathway Proteins in Thrombin-Stimulated Platelets and Regutates Platelets Activation in Vitro]

Cong-Chao Qiao; Pan Sun; Xin Yuan; Na Su; Peng Jiang; Fang-Zhao Lin

doi:10.19746/j.cnki.issn.1009-2137.2023.04.033

[Activation of Hippo Pathway Proteins in Thrombin-Stimulated Platelets and Regutates Platelets Activation in Vitro]

Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2023;31(4):1143-1149. doi: 10.19746/j.cnki.issn.1009-2137.2023.04.033.

[Article in Chinese]

Authors

Cong-Chao Qiao¹, Pan Sun¹, Xin Yuan¹, Na Su¹, Peng Jiang², Fang-Zhao Lin³

Affiliations

¹ Institute of Blood Transfusion, Chinese Academy of Medical Science & Peking Union Medical College, Chengdu 610052, Sichuan Province, China.
² Institute of Blood Transfusion, Chinese Academy of Medical Science & Peking Union Medical College, Chengdu 610052, Sichuan Province, China,E-mail: peng_jiang2014@163.com.
³ Institute of Blood Transfusion, Chinese Academy of Medical Science & Peking Union Medical College, Chengdu 610052, Sichuan Province, China,E-mail: lfangzhao@126.com.

PMID: 37551490
DOI: 10.19746/j.cnki.issn.1009-2137.2023.04.033

Abstract
in English, Chinese

Objective: To investigate the phosphorylation levels of Hippo pathway proteins in thrombin stimulated platelets and to explore its effects on platelet activation.

Methods: The phosphorylation levels of Hippo pathway proteins - Mammalian STE20-like kinase 1/2 (MST1/2), Nuclear Dbf2 related kinase 1/2 (NDR1/2) and Mps one binder 1(MOB1) in human thrombin stimulated platelets was detected by Western blot. The effect of MST1/2 inhibitor XMU-MP-1 on platelet aggregation was detected by Platelet Aggregometer. The effect of XMU-MP-1 on platelet integrin αIIbβ3 activation and CD62p expression was detected by flow cytometry. The effect of XMU-MP-1 on the "outside-in" signal of platelet integrin was detected by blood clot retraction test. The effects of XMU-MP-1 on platelet Hippo pathway proteins and p38 phosphorylation levels was detected by Western blot.

Results: The phosphorylation levels of MST1/2, NDR1/2 and MOB1 were significantly increased in thrombin activated platelets; XMU-MP-1 inhibited thrombin-induced platelet aggregation and αIIbβ3 activation, but did not affect α-granules release and clot retraction. In addition, thrombin induced phosphorylation of the Hippo proteins were decreased in XMU-MP-1 treated platelets, while the phosphorylation of p38 was not affected.

Conclusion: In thrombin stimulated platelets, Hippo pathway proteins were activated and contributed to platelets activation.

题目: Hippo通路蛋白在凝血酶活化的血小板中激活并调控体外血小板活化.

目的: 探讨Hippo通路蛋白在凝血酶刺激的血小板中的磷酸化水平，并探索其对血小板活化的影响.

方法: 免疫印迹法检测凝血酶活化的人血小板中Hippo通路蛋白—哺乳动物STE20样激酶1/2（MST1/2）、核Dbf2相关激酶1/2（NDR1/2）和MOB1的磷酸化水平；血小板聚集仪检测MST1/2抑制剂XMU-MP-1对血小板聚集的影响；流式细胞仪检测XMU-MP-1对血小板整合素αIIbβ3活化和CD62p表达的影响；血块回缩实验检测XMU-MP-1对血小板整合素“outside-in”信号的影响；免疫印迹法检测XMU-MP-1对血小板Hippo通路蛋白和p38磷酸化水平的影响.

结果: MST1/2、NDR1/2和MOB1的磷酸化水平在凝血酶活化的血小板中显著升高（P<0.05）；XMU-MP-1可抑制凝血酶诱导的血小板聚集和αIIbβ3活化（P<0.05），但不影响α颗粒释放和血块回缩；在XMU-MP-1处理的血小板中，凝血酶诱导的Hippo蛋白的磷酸化水平下降（P<0.05），而p38的磷酸化不受影响.

结论: Hippo通路蛋白在凝血酶诱导的血小板中被激活，并参与血小板活化调控.

Keywords: XMU-MP-1; hippo pathway; mammalian STE20-like kinase 1/2; nuclear Dbf2 related kinase 1/2; mps one binder 1; platelets activation.

Publication types

English Abstract

Abstract in English, Chinese

Publication types

Abstract
in English, Chinese