神经干细胞移植治疗大鼠脑缺血再灌注损伤实验研究

目的探讨大鼠胚胎神经干细胞移植治疗局灶性脑缺血再灌注损伤的可行性。方法孕龄8~10d的大鼠神经干细胞在体外扩增后,用免疫组织化学方法分别检测神经干细胞及其分化后代的特异性标志蛋白nestin、胶质纤维酸性蛋白(GFAP)和神经元特异性烯醇化酶(NSE)的表达。分别于缺血后不同时间窗将神经干细胞移植到局灶性脑缺血大鼠模型的缺血半暗带和梗塞中心,移植4w后比较不同移植部位神经干细胞存活、增殖和迁移的差异。结果从胎鼠中成功培养出悬浮生长的可表达nestin的神经球,其在含血清条件下可分化为表达GFAP的胶质细胞和表达NSE的神经元。神经干细胞移植4w后可见所有移植动物的细胞都存活,梗塞中心移植的细胞存活、增殖水平明显低于半暗带移植的细胞。结论大鼠胚胎神经干细胞移植到局灶性脑缺血再灌注损伤大鼠梗塞中心和半暗带均可长期存活,其增殖能力与移植部位密切相关。

The experimental study on treatment of cerebral ischemic-reperfusion injury with neural stem cells transplantation in rats

Objective To investigate the feasibility of treatment of focal cerebral ischemic-reperfusion injury with the rat embryonic neural stem cells (NSCs) transplantation in the rat models.Methods After proliferating to 5 passages in vitro, the neural stem cells were implanted into the ischemic region or penumbra of focal cerebral ischemic-reperfusion injury rat model. Immunohistochemistry was used to detect the expression of nestin, glial fibrillary acidic protein (GFAP) and neuron-specific enolase (NSE), which were the specific markers of neural stem cells, astrocytes and neurons respectively. The 5-bromodeoxy-uridine (BrdU) labeling method was used to mark the transplanted neural stem cells and their offsprings. The differences of survival, proliferation and migration of the neural stem cells in different spatial regions were observed at 4 w after transplantation .Results The NSCs from embryonic rat were successfully cultured which formed typical neurospheres in suspension and expressed nestin; on serum condition the NSCs differentiated into astrocytes expressing GFAP and neurons expressing NSE. The NSCs could live in all the animals at 4 w after they were transplanted into the focal cerebral ischemic-reperfusion injury rat models. More NSCs proliferated in the penumbra than that in the core of infarct region.Conclusion The rat embryonic NSCs can live in a long period and migrate widespread after they are transplanted into the ischemic region or penumbra of focal cerebral ischemic -reperfusion injury rat models and their capacity of proliferation is related to the transplanted region.