氯化钴模拟法构建骨骼肌细胞缺氧模型  

Construction of a NewModel for SkeletalMuscle Cell Hypoxia Using Cobalt Chloride

作　　者：李晰[1] 辛世杰[1] 王磊[1] 宋泽[1] 荆玉辰[1] 曹辉[1] 段志泉[1] 张健[1] 

机构地区：[1]中国医科大学附属第一医院普外教研室血管甲状腺外科,沈阳110001

出　　处：《中国医科大学学报》2014年第3期265-268,共4页Journal of China Medical University

基　　金：国家自然科学基金(81170295)

摘　　要：目的观察CoCl2对体外培养的原代骨骼肌细胞的影响,探讨合理的体外化学模拟低氧模式。方法利用组织贴块法培养并鉴定原代骨骼肌细胞,给予CoCl2处理,观察不同时间和浓度细胞凋亡情况,以及通过免疫印迹及实时定量聚合酶链反应检测低氧诱导因子1α(HIF-1α)、BAX的表达变化情况。结果与对照组比较,CoCl2处理组的细胞存活率明显下降(P<0.05),凋亡率明显增加(P<0.05),且骨骼肌细胞存活率下降程度及细胞凋亡率增加程度均随着氯化钴浓度增加时间延长而升高。HIF-1α、BAX蛋白水平亦随CoCl2浓度升高和时间延长而增加。结论 CoCl2对原代骨骼肌细胞的增殖凋亡及相关基因的表达成呈时间和浓度依赖性,成功建立起一种体外培养原代骨骼肌细胞缺氧诱导模型,可以作为体外研究骨骼肌缺血缺氧的良好模型。Objective To investigate the effect of cobalt chloride on primary cultured skeletal muscle cells and explore the reasonable strategy of simulation chemical hypoxia by using CoC12. Methods Primary skeletal muscle cells were isolated and cultured via tissue explants adhesion meth- od, and identified by immunofluorescence assay. Cells were then incubated with different concentration of cobalt chloride for different time length, and the cell apoptosis was detected respectively. Immunoblotting and qRT-PCR were used to determine the mRNA and protein levels of HIF- 1 α and BAX. Results Compared with control group, cell survival rate was significantly decreased （P 〈 0.05 ）, and apoptotic rate was markedly increased in cells treated with COCl22 （P 〈 0.05 ）. Moreover, the effects were positively correlated with the increasing concentration and exposure time of COCl2. Conclusion Effect of COCl2 on the primary skeletal muscle cells proliferation and apoptosis is dose and time-dependent. An in vitro hypoxia model of skeletal muscle cell was successfully established, which could be a good model for investigating ischemia/hypoxia on skeletal muscle cells.

关 键 词：氯化钴 骨骼肌细胞 凋亡 低氧诱导因子1Α 

分 类 号：R654.3[医药卫生—外科学]