【作者】 陈有； 冯世庆； 吴秋丽； 周先虎； 宁广智； 刘举； 冯洪永； 周恒星；

【Author】 CHEN You,FENG Shi-qing*,ZHOU Xian-hu et al.Department of Orthopaedics,Tianjin Medical University General Hospital,Tianjin 300052,China

【机构】 天津医科大学总医院；

【摘要】 目的观察人雪旺细胞(1hSCs)对人脐带间充质干细胞(hUC-MSCs)在脊髓损伤(SCI)体内存活、分化的影响,通过分子生物学技术检测胶质纤维酸性蛋白(GFAP)、髓磷脂碱性蛋白(MBP)与高分子量神经丝蛋白(NF-H)的表达,描述其变化的时间特征。方法分离、培养及纯化hUC-MSCs和hSCs。取8周龄雌性Wistar大鼠60只,以Impactor Model-Ⅱ型打击器制作脊髓胸10(T10)损伤模型。实验随机均分为4组:DMEM对照组(A),hSCs移植组(B),hUC-MSCs移植组(C),hUC-MSCs与hSCs联合移植组(D),各组分别于移植后1、2、3、4周取材。结果免疫荧光染色显示,在人细胞核特异性抗体(MAB1281)染色阳性基础上,D组可见MBP与NF-H染色阳性细胞,其它各组为阴性。分子生物学检测显示,A组3种神经标志物在蛋白与mRNA水平的表达量随时间延长均逐渐增高,且GFAP在移植后2周明显增高,而MBP与NF-H在移植后3周明显增高。与对照组相应时间点比较,移植组MBP、NF-H的表达量较高而GFAP的表达量较低,其中D组的MBP、NF-H的表达量最高,差异有统计学意义(P<0.05),但D组GFAP各时间点的表达量差异无统计学意义(P>0.05)。结论 hSCs可促进hUC-MSCs向神经元与少突胶质细胞方向分化并增强其对胶质瘢痕的抑制作用。更多还原

【Abstract】 Objective To discuss the influence of human Schwann Cells(hSCs) on the survival and differentiation of human umbilical cord mesenchymal stem cells(hUC-MSCs) in Wistar rats with spinal cord injury(SCI);To detect the expression of Glial fibrillary acidic protein(GFAP),Myelin basic protein(MBP) and High molecular weight neurofilament(NF-H),describe their changes of the time-dependent characteristic. Methods Firstly,the hUC-MSCs and the hSCs were isolated and purified in vitro.Sixty 8-week-old female Wistar rats were subjected to TIO SCI by Impactor Model-Ⅱtype device and divided into four groups randomly and evenly:DMEM control group(A),hSCs transplantation group(B),hUC-MSCs transplantation group(C),co-transplantation group of hSCs and hUC-MSCs(D),The injured spinal cord tissues were taken out at 1,2,3,4 weeks after transplantation. Results Immunofluorescence staining showed that,on the basis of the MAB1281 positive staining,the MBP and NF-H positive cells can be seen in group D,while other groups were negative.The statistical analysis of Western-Bolt and Real-Time PCR indicate that the three markers were gradually increased with time,and the expression of GFAP was significantly increased at the 2nd weeks following transplantation while other two kinds of markers were significantly increased at the 3rd weeks.Compared with the group A of the corresponding time point,both the expression of MBP and NF-H were higher in group B,C and D,while the expression of GFAP were lower,more precisely,the expression of MBP,NF-H were the highest in group D, the difference was statistically significant(P <0.05);However,there was no significant change of the GFAP expression at each time point in group D(P> 0.05).Conclusions hUC-MSCs can differentiate into neurons and oligodendrocyte with the effect of hSCs,and its inhibitory effect on the formation of glial scar were enhanced.更多还原