ROCK1及其相关信号分子参与张应变调控的血管平滑肌细胞增殖
发布时间：2018-10-16 19:02
【摘要】：目的探讨Rho相关卷曲蛋白激酶1(Rho-associated coiled-coil containing protein kinase 1,ROCK1)及其相关信号分子在感受张应变机械刺激、调控血管平滑肌细胞(vascular smooth muscle cells,VSMCs)增殖功能中的作用。方法应用张应变加载系统对体外培养VSMCs施加牵张幅度10%、频率1.25 Hz生理性周向张应变;Brdu检测VSMCs增殖水平;Western blotting检测力学加载后VSMCs的ROCK1表达水平以及蛋白激酶C(protein kinase C,PKC)α/βII、蛋白激酶D(protein kinase D,PKD)、胞外信号调节激酶(extracellular regulated protein kinase,ERK)磷酸化水平;采用RNA干扰技术(RNA interference,RNAi)检测ROCK1对VSMC增殖和PKCα/βII、PKD、ERK磷酸化的调控作用。结果 10%生理性张应变加载12、24 h显著抑制VSMCs的ROCK1表达,并显著抑制PKD和ERK的磷酸化;10%生理性张应变加载12 h显著抑制PKCα/βⅡ的磷酸化,但加载24 h PKCα/βⅡ的磷酸化与静止对照组相比无显著差异。RNAi抑制VSMCs的ROCK1表达后,VSMCs增殖水平显著降低,同时PKCα/βⅡ和PKD磷酸化水平显著降低,但ERK磷酸化无明显变化。结论 10%生理性张应变可能通过抑制ROCK1表达调控PKCα/βⅡ和PKD的磷酸化水平,从而影响VSMCs增殖,维持血管稳定性。探讨张应变力学刺激调控血管细胞功能的细胞内信号转导网络,对心血管生理和疾病病理机制研究具有一定意义。
[Abstract]:Objective to investigate the role of Rho associated crimp protein kinase 1 (Rho-associated coiled-coil containing protein kinase 1 / ROCK1) and its associated signaling molecules in the proliferation of vascular smooth muscle cells (VSMCs) by sensing tensile strain mechanical stimulation and regulating the proliferation of VSMCs. Methods tension strain loading system was applied to VSMCs cultured in vitro with strain amplitude of 10 and frequency 1.25 Hz physiologically circumferential tensile strain. Brdu was used to detect VSMCs proliferation level; Western blotting was used to detect ROCK1 expression of VSMCs after mechanical loading and protein kinase C (protein kinase CpPKC) 伪 / 尾 II, eggs were detected by Brdu. The phosphorylation level of white kinase (D (protein kinase) and extracellular signal-regulated kinase (extracellular regulated protein kinase,ERK); The effects of ROCK1 on VSMC proliferation and PKC 伪 / 尾 II,PKD,ERK phosphorylation were detected by RNA interference technique (RNA interference,RNAi). Results the ROCK1 expression of VSMCs and the phosphorylation of PKD and ERK were significantly inhibited by 10% physiological strain loading for 12h, and the phosphorylation of PKC 伪 / 尾 鈪，
本文编号：2275340
[Abstract]:Objective to investigate the role of Rho associated crimp protein kinase 1 (Rho-associated coiled-coil containing protein kinase 1 / ROCK1) and its associated signaling molecules in the proliferation of vascular smooth muscle cells (VSMCs) by sensing tensile strain mechanical stimulation and regulating the proliferation of VSMCs. Methods tension strain loading system was applied to VSMCs cultured in vitro with strain amplitude of 10 and frequency 1.25 Hz physiologically circumferential tensile strain. Brdu was used to detect VSMCs proliferation level; Western blotting was used to detect ROCK1 expression of VSMCs after mechanical loading and protein kinase C (protein kinase CpPKC) 伪 / 尾 II, eggs were detected by Brdu. The phosphorylation level of white kinase (D (protein kinase) and extracellular signal-regulated kinase (extracellular regulated protein kinase,ERK); The effects of ROCK1 on VSMC proliferation and PKC 伪 / 尾 II,PKD,ERK phosphorylation were detected by RNA interference technique (RNA interference,RNAi). Results the ROCK1 expression of VSMCs and the phosphorylation of PKD and ERK were significantly inhibited by 10% physiological strain loading for 12h, and the phosphorylation of PKC 伪 / 尾 鈪，
本文编号：2275340
本文链接：https://www.wllwen.com/yixuelunwen/swyx/2275340.html
