论文部分内容阅读
应用Ｕｌｔｉｍａ型激光共聚焦扫描显微镜、ｆｌｕｏ－３荧光探针，检测大鼠脑微血管内皮细胞在正常体外培养及缺氧培养时游离钙含量的动态变化。缺氧由培养液中加入２ｍｍｏｌ连二亚硫酸钠（Ｎａ２Ｓ２Ｏ４）同时充入Ｎ２（９５％）、ＣＯ２（５％）气体造成。实验结果发现，当缺氧１～２ｍｉｎ时，内皮细胞Ｃａ２＋含量迅速增加，２～３ｍｉｎ达高峰。此后，尽管内皮细胞仍处于缺氧条件，荧光强度却逐渐减弱。停止缺氧后，荧光强度曲线可趋于稳定。应用Ｎｉｋｏｎ倒置相差显微镜观察内皮细胞形态学变化，发现缺氧３～５ｍｉｎ，内皮细胞发生肿胀。缺氧８～１０ｍｉｎ，细胞表面有明显受损迹象。推测其形态学的变化与胞内Ｃａ２＋含量的增高有关
Ultima confocal laser scanning microscopy and fluo-3 fluorescent probe were used to detect the dynamic changes of free calcium content in rat cerebral microvascular endothelial cells during normal in vitro culture and hypoxia culture. Hypoxia was caused by the addition of 2 mmol of sodium dithionite (Na2S2O4) to the culture medium while filling N2 (95%) and CO2 (5%). The experimental results showed that when hypoxia for 1 ~ 2min, the content of Ca2 + in endothelial cells increased rapidly and peaked at 2 ~ 3min. Since then, although the endothelial cells are still under hypoxic conditions, the fluorescence intensity is gradually reduced. After stopping the hypoxia, the fluorescence intensity curve can be stabilized. The morphological changes of endothelial cells were observed by Nikon inverted phase contrast microscope and found that the endothelial cells swelled with hypoxia for 3-5 minutes. Hypoxia 8 ~ 10min, cell surface obvious signs of damage. Speculated that its morphological changes and increased intracellular Ca2 + content