摘要
背景：突触素P38广泛存在于机体所有神经终末，与神经生长、修复、再生和突触重塑密切相关。目的：实验假设细胞移植及电刺激治疗对突触素P38具有调节作用，拟验证这种作用对局灶性脑缺血大鼠突触素P38表达的影响，设计、时间及地点：随机对照动物实验及细胞观察，于2005—0812007—02在佳木斯大学神经科学研究所完成。材料：清洁级8周龄Wistar大鼠96只，随机分为神经干细胞组、电刺激＋神经干细胞组、共移植体组、电刺激＋共移植体组，24只，组。另取新生24h内Wistar鼠和1～7d龄Wistar鼠各2只，分别用于神经干细胞、脑微血管内皮细胞的分离培养。方法：将传代的脑微血管内皮细胞按3×10^7L^-1密度接种在培养瓶中，贴壁后吸出脑微血管内皮细胞培养液，涂一层层粘连蛋白后接种于神经干细胞上，密度为6×10^7L^-1，待神经干细胞贴壁后吸出培养液，再涂以一层层粘连蛋白，接种3×10^8L^-1密度的脑微血管内皮细胞，共培养12～24h，将细胞从瓶壁上依次刮起，适力吹打成1~3个脑微血管内皮细胞与5～18个神经干细胞相互包绕的细胞团，过滤后筛网上的细胞团即为神经干细胞-脑微血管内皮细胞的共移植体。各组大鼠均采用线栓法建立大脑中动脉缺血模型，其中电刺激＋神经干细胞组、电刺激＋共移植体组大鼠于造模前1d行小脑顶核刺激，电流强度50μA，频率100Hz，时程0．5ms，连续刺激1h。造模后3d将培养的神经干细胞、共移植体溶解于磷酸盐缓冲液中，调整浓度为2×10^10L^-1，各组均于右侧纹状体缺血半暗带区移植对应悬液10μL。主要观察指标：移植后3，7，14．60d免疫组织化学法检测缺血区突触素P38的表达。结果：光学显微镜下突触素P38阳性细胞染色呈棕黄色点状或颗粒状沉积，分布较密集，主要分布在神经毡内，部分围绕在神经元胞体周�
BACKGROUND： Synaptophysin P38 is widely distributed in nerve ending of a body, which is closely associated with nerve growth, recovery, regeneration and synapsis remodeling. OBJECTIVE： It is supposed that cell transplantation and electro-stimulant therapy have a regulatory effect on synaptophysin P38. This study served to verify the regulatory effect of synaptophysin P38 on focal cerebral ischemia rats. DESIGN, TIME AND SETTING： The randomized control animal experiment and cell observation were performed at the Institute of Neuroscience, Jiamusi University from August 2005 to February 2007. MATERIALS： Totally 96 clean Wistar rats aged 8 weeks were randomly divided into a neural stem cell group, an electric stimulation plus neural stem cell group, a co-transplant group and a electric stimulation plus co-transplant group, with 24 in each group. An additional two Wistar rats aged less than 24 hours and two Wistar rats aged 1-7 days were used for isolation and culture of neural stem cells and cerebral microvascular endothelial ceils. METHODS： After passage, cerebral microvascular endothelial cells were incubated in a culture flask at a density of 3 × 10^7 L^-1. After adherence, cerebral microvascular endothelial cell medium was removed. After laminin overlay, cerebral microvascular endothelial cells were incubated in neural stem cells at the density of 6×10^8 L^-1. After adherence of neural stem ceils, the medium was removed. After laminin overlay, cerebral microvascular endothelial cells were incubated at the density of 3 × 10^7 L^-1. Coculture was performed for 12 24 hours. Cells were collected and blown into cell masses of 1 3 cerebral microvascular endothelial cells and 5-18 neural stem cells. After filtering, cell masses on the grit were co-transplant of neural stem cells and cerebral microvascular endothelial ceils. Rat models of middle cerebral artery occlusion （MCAO） were established by intraluminal vascular occlusion. Rats in the electric stimulation plus neural stem cell group and electric
出处
《中国组织工程研究与临床康复》
CAS
CSCD
北大核心
2008年第34期6621-6625,共5页
Journal of Clinical Rehabilitative Tissue Engineering Research
基金
国家自然科学基金(30450062)
黑龙江省自然基金重点项目(ZJY0508)~~
