摘要
目的观察脂多糖 (Lipopolysaccharid ,LPS)和肿瘤坏死因子α(Tumornecrosisfactor -α ,TNF -α)对血脑屏障内皮细胞 (Blood_brainbarrierendothelialcell,BBBEC)胞浆内钙离子浓度(plasmicinnerconcentrationofcalcium ,[Ca2 +]i)的影响。方法用Fluo -3/AM做为荧光探针对钙离子进行负载 ,激光扫描共聚焦显微镜观察不同浓度LPS和TNF -α刺激下BBBEC胞浆内钙离子浓度的变化。结果在TNF -α刺激下 ,单个BBBEC[Ca2 + ]i呈浓度依赖性的一过性升高。不同浓度TNF -α引起的BBBECCa2 + 的荧光强度达高峰的时间基本一致 ,高峰荧光强度和荧光持续时间随TNF -α浓度的升高而增加。低浓度LPS(1μg/ml和10μg/ml)刺激细胞时 ,BBBEC[Ca2 +]i无明显变化 ,高浓度LPS刺激细胞时 ,可见[Ca2 +]i呈短暂性升高 ,随后下降。结论TNF -α和LPS在极短的时间内导致BBBEC[Ca2 + ]i的升高 ,激活BBBEC ,这一过程是缺血性脑损伤极早期的重要病理机制之一。
Objective To observe the change of plasmic inner concentration of calcium(i)of blood_brain barrier endothelial cell(BBBEC)after stimulation by lipopolysaccharid(LPS)and tumor necrosis factor-α(TNF-α).Methods The cultured BBBECs were loaded with Fluo-3/AM.After stimulation by TNF-αand LPS,confocal laser scanning microscope was used to measure i.Results i of single BBBEC increased temporally after stimulation by TNF-α.Though the peak time of fluorescence intensity was all about50s after stimulation,the maximum fluorescence intensity and lasting time of fluorescence increased with increasing dosage of TNF-α).High concentraion LPS could induce temporal increase of i while lowconˉcentration LPSinduced no significant change.Conclusions TNF-αand LPS cause rapid increase of BBBEC i,which might act as one of the important mechanisms in the early phase of brain ischemic injury.
出处
《中国微循环》
2003年第1期14-16,共3页
Journal of Chinese Microcirculation
基金
国家自然科学基金资助项目(39670267)
军队医药卫生基金项目(96M123)
