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 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 后显著减少了划痕愈合面积，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 的蛋白表达显著降低。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 的蛋白表达，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 后显著诱导了 p-AMPK 的蛋白表达，孵育 24 h 后显著抑制了 VSMCs 迁移；提示p-AMPK的蛋白表达参与调控VSMCs 迁移。结论 生理性周期性张应变能通过激活 p-AMPK 的蛋白表达进而抑制 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 The aim of the current study is to explore the role of 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 of 10% amplitude and 1.25 Hz frequency to primary rat VSMCs, to simulate the mechanical stimulation of VSMCs in vivo; The protein expression of p-AMPK in VSMCs was detected by Western blotting; 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, suggesting 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 can 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, suggesting that the VSMC migration regulated by physiological cyclic stretch is of great significance for maintaining vascular homeostasis.