生理性周期性张应变通过激活AMPK磷酸化抑制血管平滑肌细胞迁移  

Physiological Cyclic Stretch Inhibits Vascular Smooth Muscle Cell Migration via Activating AMPK Phosphorylation

作　　者：范洋晶 李之音 姜宗来[1] 齐颖新[1] 韩悦[1] FAN Yangjing;LI Zhiyin;JIANG Zonglai;QI Yingxin;HAN Yue(Institute of Mechanobiology and Medical Engineering,School of Life Sciences 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》2022年第2期335-341,共7页Journal of Medical Biomechanics

基　　金：国家自然科学基金项目(12072197,11972232)。

摘　　要：目的探讨细胞能量代谢的关键调节因子AMP激活的蛋白激酶AMPK在血管平滑肌细胞(vascular smooth muscle cells,VSMCs)响应生理性周期性张应变力学刺激后对VSMCs迁移的影响。方法采用Flexcell-5000T体外细胞张应变加载系统,对大鼠原代培养的VSMCs施加10%幅度、1.25 Hz频率的周期性张应变,模拟VSMCs在体内的生理性力学环境;以未加载周期性张应变的静态细胞为对照组,Western blotting检测VSMCs的p-AMPK蛋白表达;划痕实验检测VSMCs迁移功能。结果与静态组的细胞相比,生理性周期性张应变加载24 h后显著减少划痕愈合面积,提示生理性周期性张应变抑制VSMCs迁移;生理性周期性张应变加载3 h后,VSMCs的p-AMPK蛋白表达显著升高,而加载24 h后p-AMPK蛋白表达显著降低。在生理性周期性张应变加载条件下,孵育AMPK抑制剂可以在张应变加载3 h后显著降低p-AMPK蛋白表达,而在张应变加载24 h后显著促进VSMCs迁移;在静态条件下孵育AMPK激活剂AICAR 3 h后显著诱导p-AMPK蛋白表达,孵育24 h后显著抑制VSMCs迁移;提示p-AMPK蛋白表达参与调控VSMCs迁移。结论生理性周期性张应变能通过激活p-AMPK蛋白表达,进而抑制VSMCs迁移,提示生理性周期性张应变调控VSMCs迁移对维持血管稳态具有重要意义。Objective To explore the role of adenosine monophosphate-activated protein kinase(AMPK),a key regulator of cellular energy metabolism,in vascular smooth muscle cell(VSMC)migration in response to physiological cyclic stretch.Methods The Flexcell-5000T mechanical loading system was applied with a physiological cyclic stretch at 10%amplitude and 1.25 Hz frequency to primary rat VSMCs,to simulate mechanical stimulation of VSMCs in vivo.The protein expression of p-AMPK in VSMCs was detected by Western blotting,and VSMC migration was detected by wound healing test.Results Compared with the static group,physiological cyclic stretch loading for 24 h significantly decreased the area of wound healing,indicating that physiological cyclic stretch inhibited VSMC migration.The protein expression of p-AMPK in VSMCs was increased significantly after physiological cyclic stretch loading for 3 h,and was decreased significantly after 24 h.Under physiological cyclic stretch loading conditions,incubating AMPK inhibitor could significantly reduce the protein expression of p-AMPK after 3 h,and promote VSMC migration after 24 h;incubating AMPK activator AICAR under static conditions significantly increased the protein expression of p-AMPK after 3 h,and weakened VSMC migration after 24 h.Conclusions Physiological cyclic stretch inhibits VSMC migration by increasing the protein expression of p-AMPK,indicating that VSMC migration regulated by physiological cyclic stretch is of great significance for maintaining vascular homeostasis.

关 键 词：周期性张应变 力学刺激 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]