[1]何秀娟,许蜀闽,张涛,等.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[J].第三军医大学学报,2007,29(15):1477-1481.
 HE Xiu-juan,XU Shu-min,ZHANG Tao,et al.Dynamic changes of membrane potentials, cytoplasmic and mitochondrial Ca2+ in cultured pulmonary artery smooth muscle cells exposed to hypoxia[J].J Third Mil Med Univ,2007,29(15):1477-1481.
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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(/HTML )
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
29卷
期数:
2007年第15期
页码:
1477-1481
栏目:
论著
出版日期:
2007-08-15

文章信息/Info

Title:
Dynamic changes of membrane potentials, cytoplasmic and mitochondrial Ca2+ in cultured pulmonary artery smooth muscle cells exposed to hypoxia
作者:
何秀娟许蜀闽张涛李雪辉孙玮王培勇
第三军医大学高原军事医学系病理生理学与高原生理学教研室,全军高原生理与高原病研究重点实验室
Author(s):
HE Xiu-juan XU Shu-min ZHANG Tao LI Xue-hui SUN Wei WANG Pei-yong
Department of Pathophysiology and High Altitude Physiology, College of High Altitude Military Medicine, Third Military Medical University, Chongqing 400038, China
关键词:
膜电位缺氧肺动脉平滑肌线粒体
Keywords:
membrane potentials calcium hypoxia pulmonary artery smooth muscle cell mitochondria
分类号:
R322.121; R329.27; R364.4
文献标志码:
A
摘要:
目的    研究缺氧时肺动脉平滑肌细胞(pulmonary artery smooth muscle cells, 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 (hypoxic pulmonary vasocontriction, 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 (2 μmol/L)、Fluo-4/AM (4 μmol/L)和Rhod-5F (5 μmol/L)。利用Na2S2O4(2 mmol/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100 s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30 s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。
Abstract:
Objective    In order to understand the response of pulmonary artery smooth muscle cells (PASMCs) to hypoxia and explore the mechanism of hypoxic pulmonary vasocontriction (HPV), the changes of membrane potentials, cytoplasmic and mitochondrial Ca2+ under hypoxic stimulation were investigated.     Methods    PASMCs from adult SD rats were isolated, cultured and identified. Hypoxic stimulation was made by adding Na2S2O4 (2 mmol/L) to the incubation media. The fluorescent intensity of cell membrane potential, cytoplasmic and mitochondrial Ca2+was measured under confocal microscopy after PASMCs loaded with the fluorescent indicators Di-8-ANEPPS (2 μmol/L), Fluo-4/AM (4 μmol/L) and Rhod-5F (5 μmol/L) respectively. Each sample was observed for 10 min.     Results    After Na2S2O4stimulation, the fluorescent intensity of PASMCs membrane potentials recovered following a transient significant decrease, increased gradually to the climax at 100 s and then decreased gradually to the level lower than the initial value. The fluorescent intensity of cytoplasmic Ca2+ increased gradually. The fluorescent intensity of mitochondrial Ca2+ increased transiently and lasted about 30 s but then decreased below the initial value, and followed by a steady increase above the initial value.     Conclusion    Under hypoxic stimulation, the membrane potential, cytoplasmic and mitochondrial Ca2+ changed dynamically in specific patterns. In addition to disturbance of cytoplasmic Ca2+, disturbance of mitochondrial Ca2+ might play important roles in the development of HPV.

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更新日期/Last Update: 2008-09-26