摘要
目的:探讨溶血卵磷脂(LPC)对牛视网膜微血管内皮细胞(BRECs)凋亡及细胞内钙离子浓度的影响。方法:原代分离培养牛视网膜微血管内皮细胞,加入不同浓度的LPC,分别培养6h、12h、24h,通过吖啶橙染色在荧光显微镜下观察细胞凋亡,流式细胞检测细胞凋亡率,F2000荧光光度计测细胞内钙离子浓度,并计算游离钙浓度。结果:(1)60μmol/LLPC作用牛视网膜微血管内皮细胞24h后,细胞凋亡率明显高于LPC浓度为20μmol/L及40μmol/L时;(2)LPC浓度从20μmol/L增加到60μmol/L过程中,细胞内游离钙离子浓度明显增加,而且细胞内游离钙离子浓度的增加与LPC的作用时间有关,作用时间越长,钙离子浓度越高。结论:LPC能增加细胞内游离钙离子浓度,促使BRECs的凋亡,而且LPC的这种作用具有时间和剂量依赖性。因此,脂代谢紊乱可能是通过诱导BRECs的凋亡而发挥作用的。
Objective： To study whether lysophosphatidylcholine （LPC） can induce apoptosis of bovine retinal endothelial cells （BRECs） in vitro. Methods： BRECs were exposed to LPC at a range of concentrations and cultured in 12-well plates for 6, 12, 24 hours. Apoptosis was examined on acridine orange （AO） stained specimens under fluorescence microscope. Flow cytometry was used to detect apoptosis rate. The intracellular content of calcium was determined with F-2000 type spectrotluorometer, and the concentration of free calcium ion was calculated. Results： After 24 hours, the rates of apoptosis were significantly higher in BRECs received 60μmol/L of LPC than in those that received 20 or 40μmol/L of LPC. The concentration of free calcium ion was obviously increased as the concentrations of LPC rised from 20μmol/L to 60μmol/L, and the concentration of free calcium ion was influenced by the exposure duration of BRECs to LPC, that, the longer the affecting time, the higher the concentration of free calcium ion. Conclusions： LPC has effects on the intracellular concentration of free calcium ion to induce apoptosis of BRECs in a time- and dose-dependent fashion. Lipid metabolic disturbance might function through causing apoptosis of BRECs.
出处
《贵阳医学院学报》
CAS
2006年第6期502-505,509,共5页
Journal of Guiyang Medical College
关键词
溶血卵磷脂
凋亡
BRECs
细胞内钙离子浓度
lysophosphatidylcholine
apoptosis
bovine retinal endothelial ceils
the intracellular concentration of calcium ion
