中文摘要：

目的：通过采用D-273内质网荧光分子探针来观察内质网钙库在脑缺血再灌注中的变化和作用。 方法：实验于2004—09/2005—05在中国中医科学院针灸研究所形态室完成。实验选用原代培养的8．0-9．0d的新生大鼠海马细胞，实验分正常对照组和缺血再灌注模型组，正常对照组换有糖Earle液，正常条件下培养。缺血再灌注模型组模拟缺血将培养液换成缺氧缺糖的Earle液3．5h，模拟再灌注状态换回培养液12h。应用内质网荧光分子探针D-273染色。激光共聚焦显微镜静态扫描观察缺血再灌注模型组和正常对照组的胞内荧光强度。 结果：正常对照组细胞的平均荧光值为602±131（n=10），缺血再灌注模型组细胞缺氧缺糖3．5h后再灌12h后内质网的荧光值为310±44（n=10），出现了下降的趋势，缺血再灌注模型组细胞内质网荧光值非常显著地低于对照组（P〈0．01）。结论：体外模拟缺血再灌注海马细胞内质网活性下降，损伤模型成功；D-273可作为一种特异性内质网的荧光分子探针。

英文摘要：

AIM： To observe the change and effect of endoplasmic reticulum （ER） calcium store on cerebral ischemia reperfusion （IR） injury by using the D-273 ER fluorescent molecular probes. METHODS： The experiment was accomplished in the Acupuncture and Moxibustion Institute, China Academy of Traditional Chinese Medicine from September 2004 to May 2005.The hippocampus cells, cultured primarily for 8.0-9.0 days in the newbom rats, were divided into control group and IR model group. The control cells were placed in the Earle liquid with sugar and cultured in the normal condition while the cells in the IR model group were placed in the Earle liquid at hypoxia and hypoglycemia to imitate ischemic state for 3.5 hours and retumed to the culture medium for 12 hours to model the reperfusion state. The fluorescence intensities in the hippocampus cells of both groups were observed by using laser confoeal scanning microscope and stained with D- 273 ER fluorescent molecular probe. RESULTS： The average levels of fluorescence in cells of control group were 602±131 （n=10） while the fluorescence levels of cells in the IR model group were 310±44 （n=10） at hypoxia and hypoglycemia for 3.5 hours, and presented the significantly decreased tendency. The ER fluorescence level was significantly lower in the IR model group than in the control group （P 〈 0.01）. CONCLUSION： The ER fluorescence reduce in the IR models in vitro, which means a successful model establishment; D-273 can be effective as a specific ER fluorescent molecular probe.