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目的动态观察大鼠持续性局灶脑缺血后星形胶质细胞的形态学变化,探讨星形胶质细胞的死亡方式。方法将48只SD大鼠随机分为免疫组化组、免疫组化对照组、电镜实验组和电镜实验对照组。采用颈内动脉线栓并环扎的方法建立持续性局灶脑缺血模型。免疫组化组和电镜实验组大鼠按照缺血时间(3h、6h、12h、24h、48h)平均随机分为5个亚组。对照组为术后24h取脑。免疫组化标本采用HE染色及TUNEL-GFAP双染。电镜标本在中心区、边缘区分别取材、染色后,透射电镜观察。结果随着时间的延长,脑缺血中心区及边缘区星形胶质细胞逐渐出现核肿胀、染色质分散、边聚、空泡变,核膜破溃,染色质溢出。随着缺血时间延长,中心区GFAP阳性细胞数逐渐减少,直至基本消失。缺血边缘区则可见到GFAP阳性细胞数量逐渐增多。TUNEL-GFAP双染未见阳性细胞。结论持续局灶脑缺血后,星形胶质细胞未见凋亡,推测胀亡是星形胶质细胞主要的死亡方式。
OBJECTIVE: To dynamically observe the morphological changes of astrocytes after focal cerebral ischemia in rats and explore the way of astrocyte death. Methods 48 Sprague-Dawley rats were randomly divided into three groups: immunohistochemistry, immunohistochemistry, electron microscopy and electron microscopy. A continuous focal cerebral ischemia model was established by means of internal carotid artery occlusion and cerclage. Immunohistochemistry and electron microscopy experimental group rats were randomly divided into 5 subgroups according to the ischemic time (3h, 6h, 12h, 24h, 48h). Control group for the brain 24h after surgery. Immunohistochemical specimens using HE staining and TUNEL-GFAP double staining. Electron microscopy in the central area, the marginal zone were drawn, stained, transmission electron microscopy. Results As time went by, the astrocytes in the central and peripheral regions of cerebral ischemia gradually developed nuclear swelling, chromatin dispersion, edge condensation, vacuolar change, nuclear membrane rupture and chromatin overflow. With the prolongation of ischemic time, the number of GFAP positive cells in the central area gradually decreased until it disappeared. Ischemia marginal zone can be seen gradually increased the number of GFAP positive cells. TUNEL-GFAP double staining no positive cells. Conclusion There is no apoptosis of astrocytes after focal cerebral ischemia, suggesting that astrocyte death is the main way of death.