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.
作者
范洋晶
李之音
姜宗来
齐颖新
韩悦
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)
出处
《医用生物力学》
CAS
CSCD
北大核心
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)。
关键词
周期性张应变
力学刺激
血管平滑肌细胞
细胞迁移
cyclic stretch
mechanical stimulation
vascular smooth muscle cells(VSMCs)
cell migration
