钙离子参与周期性高张应变和血小板源性微囊协同诱导的血管平滑肌细胞迁移  

Calcium Participates in Migration of VSMCS Synergistically Induced by Cyclic Stretch and Platelet-Derived Microvesicles

作　　者：高爽 李珊珊 李子通 范洋晶 刘泽 齐颖新[1] GAO Shuang;LI Shanshan;LI Zitong;FAN Yangjing;LIU Ze;QI Yingxin(Institute of Mechanobiology and Medical Engineering,School of Life Science and Biotechnology,Shanghai Jiao Tong University,Shanghai 200240,China)

机构地区：[1]上海交通大学生命科学技术学院力学生物学研究所,上海200240

出　　处：《医用生物力学》2021年第1期144-150,共7页Journal of Medical Biomechanics

基　　金：国家自然科学基金项目(11625209,31700816)。

摘　　要：目的探究周期性高张应变和血小板源性微囊(platelet-derived microvesicles,PMVs)对血管平滑肌细胞(vascular smooth muscle cells,VSMCs)迁移功能的影响,以及Ca^(2+)在其中的作用。方法应用FX-5000T应变加载系统对体外培养VSMCs施加5%、15%幅度周期性张应变,分别模拟VSMCs受到生理、高血压条件下的周期性高张应变;使用划痕实验检测VSMCs迁移;使用无Ca^(2+)培养基创造细胞外无游离Ca^(2+)条件;应用IP3R拮抗剂2-APB阻断细胞内Ca^(2+)释放;施加TRPV4通道拮抗剂GSK219和L型电压门控钙通道的抑制剂Nifedipine分别阻断相应Ca^(2+)通道;体外施加凝血酶激活血小板产生PMVs以模拟高血压病理环境。结果与5%周期性张应变相比,15%周期性张应变可以显著促进VSMCs迁移。去除细胞外Ca^(2+)可以抑制VSMCs迁移,但GSK219和Nifedipine对于15%高张应变诱导的VSMCs迁移无显著作用;2-APB可以抑制15%高张应变诱导的VSMCs迁移过程。同时PMVs可以显著促进15%高张应变诱导的VSMCs迁移,并且细胞外和细胞内Ca^(2+)均参与其中。结论细胞内钙和细胞外钙在15%周期性高张应变诱导的VSMCs迁移中均发挥重要作用,并且PMVs可以协同参与到上述过程中。研究结果为高血压病理性张应变诱导血管重建的分子机制和临床治疗提供力学生物学新思路。Objective To investigate the synergistic effects of pathologically elevated cyclic stretch and plateletderived microvesicles( PMVs) on migration of vascular smooth muscle cells( VSMCs) and the potential role of calcium in this process. Methods The FX-5000 T strain loading system was used to apply cyclic stretch to VSMCs with magnitudes of 5% and 15%,which simulated physiological and hypertensive situation respectively in vitro;wound healing assay was used to analyze VSMCs migration;Ca^(2+)-free medium was used to remove extracellular calcium;2-APB( an antagonist of IP3 R) was used to inhibit the release of intercellular stored calcium;GSK219( an antagonist of TRPV4) and Nifedipine( an inhibitor of L-type voltage-gated calcium channel) were applied to block the activity of respective calcium channel;thrombin was used to stimulate platelets in vitro which simulated the hypertensive activation of PMVs in vivo. Results Compared with 5% cyclic stretch,15% cyclic stretch significantly promoted VSMC migration. Removal of extracellular calcium inhibited VSMCs migration,but the application of GSK219 and Nifedipine did not affect the migration up-regulated by 15% cyclic stretch;while 2-APB which inhibited the release of intracellular stored calcium could also repress VSMCs migration under 15% cyclic stretch. PMVs further promoted VSMC migration under 15% cyclic stretch condition,and both extracellular calcium and intercellular stored calcium were involved in this process. Conclusions Both intracellular and extracellular calcium play important roles in VSMC migration induced by 15% cyclic stretch, and PMVs synergistically participate in the above process. The study is aimed to provide new mechanobiological insights into the molecular mechanism and clinical targets of vascular remodeling in hypertension.

关 键 词：周期性张应变 血小板源性微囊 血管平滑肌细胞 细胞迁移 

分 类 号：R318.01[医药卫生—生物医学工程]