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缺氧时肺动脉平滑肌细胞膜电位、细胞质及线粒体Ca2+的动态变化

, PP. 1477-1481

Keywords: 膜电位,,缺氧,肺动脉,平滑肌,线粒体

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Abstract:

目的研究缺氧时肺动脉平滑肌细胞(pulmonaryarterysmoothmusclecells,PASMCs)膜电位、细胞质游离Ca2+及线粒体游离Ca2+的动态变化,探讨缺氧性肺血管收缩(hypoxicpulmonaryvasocontriction,HPV)的可能机制。方法分离成年Wistar大鼠PASMCs进行体外培养,将PASMCs分为3组:分别负载荧光指示剂Di-8-ANEPPS(2μmol/L)、Fluo-4/AM(4μmol/L)和Rhod-5F(5μmol/L)。利用Na2S2O4(2mmol/L)的强还原特性,对细胞进行化学缺氧干预,在共聚焦显微镜下观察缺氧环境下细胞膜电位、细胞质Ca2+和线粒体Ca2+的动态变化。结果用Na2S2O4对PASMCs施加缺氧刺激后,PASMCs细胞膜电位荧光强度迅速大幅度降低,继而迅速回升,于100s时升至峰值且高于初始水平,随后逐渐下降低于初始水平;缺氧刺激后PASMCs细胞质Ca2+荧光强度缓慢增高,并呈持续增高趋势;PASMCs线粒体Ca2+荧光强度在缺氧刺激后迅速增高,持续约30s后回降至低于起始值,随后开始回升,达到并逐渐高于起始值。结论缺氧刺激PASMCs,导致膜电位、细胞质Ca2+和线粒体Ca2+呈特定模式的升高和降低的动态变化,除了细胞质Ca2+稳态失衡之外,线粒体Ca2+变化可能在HPV发生中也起着重要的作用。

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