摘要
目的建立一种有效、方便的急性分离方法,用于研究生理和病理状态下血管张力及其反应性变化机制。方法采用木瓜酶、胶原酶XI两步消化法分离单个血管平滑肌细胞。结果分离出的血管平滑肌细胞数量多,细胞内钙分布均匀;在10mmol/L咖啡因的作用下,血管平滑肌细胞内钙浓度增加、细胞收缩;分离的血管平滑肌细胞膜表面光滑,容易与玻璃微电极形成高阻封接,可记录到稳定的钾电流。结论两步酶消化法所分离出的血管平滑肌细胞膜具有良好的钙激活钾通道活性,其IP3受体和Ryanodine受体具有敏感的反应性,其收缩蛋白功能良好。
Vascular smooth muscle cells (VSMCs) are crucial for studying the mechanism and regulation of vasogenic response, but so far no ideal method has been reported for their isolation. We established a VMSC isolation method based on two-step digestion, with which a large population of single smooth muscle cells could be obtained. After cell isolation, the potassium current, cytosolic calcium concentration, reactivity to caffeine were measured, and the results demonstrated that the cells were highly viable and exhibited reactivity to the vascular constrictor to meet various needs in experimental research.
出处
《南方医科大学学报》
CAS
CSCD
北大核心
2006年第7期954-958,共5页
Journal of Southern Medical University
基金
国家自然科学基金
广东省自然科学基金~~
关键词
平滑肌细胞
反应性
膜片钳
arterial smooth muscle cells
reactivity
patch clamp