周期性张应变调控血管平滑肌细胞黏附血小板微体及其在自噬中的作用
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国家自然科学基金项目(11625209, 11222223)


Cyclic Stretch Induces Adhesion of VSMCs with Platelet-Derived Microparticles and the Role in Autophagy
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    摘要:

    目的 探讨周期性张应变力学刺激对血管平滑肌细胞(vascular smooth muscle cells,VSMCs)与血小板微体(platelet-derived microparticles, PMPs)黏附能力的影响,以及黏附的PMPs对VSMCs自噬的调控作用。方法 应用 FX-5000T 张应变加载系统,对体外培养VSMCs施加5%幅度的生理性张应变和15%幅度的高张应变;应用流式细胞术检测不同张应变作用的VSMCs与PMPs的黏附;免疫荧光检测PMPs刺激24 h后自噬标志分子微管相关蛋白轻链3(autophagy microtubule associated protein light chain 3,LC3)的表达水平;Western blotting 检测PMPs刺激24 h后VSMCs自噬相关蛋白(autophagy related protein,Atg)的表达水平。结果 与5%生理性张应变加载相比,15%高张应变加载24 h能显著增强VSMCs与PMPs的黏附水平,提示高张应变促进PMPs与VSMCs的黏附。免疫荧光和Western blotting结果显示,PMPs刺激可显著上升VSMCs中自噬标志蛋白LC3表达,同时Western blotting 检测到PMPs刺激后Atg5、Atg7、Atg12蛋白表达水平显著上升。结论 高张应变可以促进VSMCs黏附PMPs,黏附的PMPs可能通过增加Atg5、Atg7、Atg12、LC3表达,从而增强VSMCs自噬。

    Abstract:

    Objective To investigate the effect of cyclic stretch on adhesion of vascular smooth muscle cells (VSMCs) with platelet-derived microparticles (PMPs), and the role of PMPs in VSMC autophagy. Methods Cyclic stretch with the magnitude of 5% (simulating physiological mechanical stretch) or 15% (simulating pathological mechanical stretch) was subjected to VSMCs in vitro by using FX-5000T cyclic stretch loading system, and the adhesion of PMPs in VSMCs was detected by using flow cytometry. Immunofluorescence was used to detect the expression of autophagy microtubule associated protein light chain 3 (LC3) after 24 h stimulation with PMPs. Western blotting was used to detect the expression of autophagy related protein (Atg) in VSMCs after 24 h stimulation by PMPs. Results Compared with 5% cyclic stretch, 15% cyclic stretch significantly increased the adhesion ability of VSMCs with PMPs. Immunofluorescence and Western blotting result revealed that PMPs stimulation significantly increased the expression of autophagy marker protein LC3 in VSMCs. Furthermore, the protein expressions of Atg5, Atg7 and Atg12 were all significantly increased in VSMCs stimulated with PMPs. Conclusions High cyclic stretch may enhance the autophagy of VSMCs by promoting the adhesion of PMPs, which will subsequently increase the expressions of Atg5, Atg7, Atg12 and LC3.

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陈远秀,包晗,阎靖,肖倩,齐颖新.周期性张应变调控血管平滑肌细胞黏附血小板微体及其在自噬中的作用[J].医用生物力学,2020,35(1):49-56

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  • 收稿日期:2019-01-18
  • 最后修改日期:2019-02-15
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  • 在线发布日期: 2020-03-03
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