嗅鞘细胞脑出血大鼠脑内移植的运动神经功能改变和形态学研究

Experimental study on olfactory ensheathing cells transplantation to intracerebral hemorrhage rat to alter motor nerve function and morphology

目的探索嗅鞘细胞脑出血大鼠脑内移植后运动神经功能及形态学改变。方法差速贴壁法培养出较高纯度的嗅鞘细胞,用PI和P75荧光双染鉴定细胞,制作30只脑出血大鼠模型,分A、B、C3组。A组10只为单纯脑出血组;B组10只为培养液移植组;C组10只为嗅鞘细胞移植组。3组大鼠分别在术前1h,术后7d、14d、21d、28d进行运动功能学评分,并于细胞移植后第4周、第8周取脑组织做冰冻切片,通过免疫荧光检测移植细胞在大鼠脑内形态学改变。结果试验中分离、培养的嗅鞘细胞经标记移植入大鼠脑内后,经免疫荧光检测证实能在血肿腔边缘分化为成熟神经元细胞和星形胶质细胞。对3组大鼠术后进行评分,细胞移植组从第21天起其功能恢复明显优于其它两组(P<0.001)。结论嗅鞘细胞在脑出血大鼠脑内能存活并分化为成熟神经元及星形胶质细胞,能促进运动神经功能恢复。

Objective The olfactory ensheathing cells（OECs） transplanted into cerebral hemorrhage mode to alter motor nerve function and the study of morphology.Methods Get high purification of olfactory ensheathing cells by using the method of the differing rates of attachment of the various cells type,use the technology of fluoresceinstain identificated the OECs by both PI and P75,make 30 rats of cerebral hemorrhage mode,and separate 3 groups.Ten rats in the A group are the blank group;Ten rats in the B group are the DPBS transplanting group;Ten rats in the C group are the OECs transplanting group.The three groups are all scored in 1 hour before operation and 7 days,14 days,21 days and 28 days after operation by Rosenberg GA method.Evaluate the contribution of OECs transplanting curing rat cerebral hemorrhage by statistics method.And get brain of rat to make frozen section in 4weeks,8weeks after operation. Detection the morphology of transplanting cells by immunofluorescence technique.Results We get in experiment transplantd in cerebral hemorrhage rats brain.We found these OECs differentiating to riped neuron cell and horizontal cell unround cavity of blood tumor by immunofluorescence way.After scoring the three groups by Rosenberg GA method and statisticing them.We found that olfactory ensheathing cells injected significantly reduced motor deficits compared with control groups which cerebral hemorrhage after 21 days （ P 〈0.001）.Conclusions The OECs survival in the brain of rat cerebral hemorrhage,then differentiating to ripe neuron cell and horizontal cell.The treatment of OECs transplanting for rat cerebral hemorrhage could promote recovery of motor nerve function.