机械敏感性离子通道Piezo1介导高静水压诱导大鼠冠脉平滑肌细胞表型改变

Role of mechanosensitive ion channel Piezo1 in mediating phenotypic changes of rat coronary arterial smooth muscle cells induced by high hydrostatic pressure

目的探究Piezo1在高静水压诱导的冠脉平滑肌细胞(CASMCs)表型改变中的作用机制。方法分别于0、120和180 mmHg压力下干预原代Wistar大鼠CASMCs 24 h,Western blot检测Piezo1、收缩表型相关蛋白Cav1.2、SM-MHC、α-SMA和合成表型相关蛋白OPN、MMP-2、Col1a1的表达,激光共聚焦检测压力对Piezo1介导的钙离子内流的影响;分别用Piezo1激动剂Yoda1、抑制剂GsMTx4处理CASMCs以及siRNA敲低Piezo1,Western blot检测收缩表型及合成表型相关蛋白的表达。结果与0 mmHg相比,120和180 mmHg干预CASMCs后,Piezo1、OPN、MMP-2、Col1a1表达增加,Cav1.2、SM-MHC、α-SMA表达减少。180 mmHg高压干预后,Piezo1介导的钙离子内流强于常压对照组,但敲低Piezo1后有所下降。在常压下,用Yoda1处理CASMCs后,收缩表型相关蛋白表达减少,合成表型相关蛋白表达增加。与对照组相比,180 mmHg下分别用GsMTx4抑制和siRNA敲低Piezo1后,收缩表型相关蛋白表达增加,合成表型相关蛋白表达减少。结论Piezo1在高静水压诱导CASMCs由收缩表型向合成表型转变中起促进作用。

Aim To investigate the mechanism of Piezo1 in the phenotypic changes of rat coronary arterial smooth muscle cells(CASMCs)induced by high hydrostatic pressure.Methods CASMCs were isolated from Wistar rats and stimulated for 24 h at 0,120 and 180 mmHg,respectively.The expressions of Piezo1,contractile phenotype-related proteins including Cav1.2,SM-MHC,α-SMA and synthetic phenotype-related proteins including OPN,MMP-2,Col1a1 were detected by Western blot.The effect of calcium influx mediated by Piezo1 was detected by Laser confocal microscopy.CASMCs were treated with Piezo1 agonist Yoda1,inhibitor GsMTx4 and Piezo1-siRNA,respectively and the expressions of contractile phenotype and synthetic phenotype-related proteins were detected by Western blot.Results Compared with control(0 mmHg),the expressions of Piezo1,OPN,MMP-2 and Col1a1 increased,but the expressions of Cav1.2,SM-MHC andα-SMA decreased in 120 mmHg as well as 180 mmHg group.After stimulated by 180 mmHg high pressure,Piezo1-mediated calcium influx was stronger than that in 0 mmHg group,but decreased after Piezo1 knockdown.Treated with Yoda1 at 0 mmHg,the expression of contractile phenotype-related protein decreased while the expression of synthetic phenotype-related protein increased compared with DMSO group.After using GsMTx4 to inhibit or siRNA to knockdown Piezo1 at 180 mmHg,the expression of contractile phenotype-related protein increased and the expression of synthetic phenotype-related protein decreased compared with the control group.Conclusion Piezo1 promotes the transition from contractile phenotype to synthetic phenotype of CASMCs induced by high hydrostatic pressure.