摘要
背景:近年来骨髓间充质干细胞移植在中枢神经系统中的应用越来越广泛,研究其最佳的移植方式具有重要意义。目的:观察不同途径移植骨髓间充质干细胞对创伤性脑损伤大鼠海马神经元的影响。方法:分离培养Wistar大鼠骨髓间充质干细胞,以PKH26标记第3代骨髓间充质干细胞,荧光显微镜观察并流式细胞仪检测标记率;采用改进Feeney法制作脑损伤致海马神经元损伤模型。60只Wistar大鼠随机分为3组,脑损伤区移植组:造模后注射含骨髓间充质干细胞的生理盐水10μL;尾静脉移植组:自尾静脉注入含骨髓间充质干细胞的生理盐水1mL;创伤性脑损伤组:不给予任何处理。结果与结论:荧光显微镜观察PKH26标记的BMSCs呈球形,呈现均匀分布的红色荧光;PKH26标记BMSCs的阳性率>99%。脑损伤区移植组大鼠学习记忆功能明显好于其他两组,PKH26阳性细胞和苏木精-伊红染色切片中的神经元数量脑损伤区移植组多于尾静脉移植组,尾静脉移植组多于创伤性脑损伤组(P<0.05)。GAP-43mRNA的表达脑损伤区移植组高于尾静脉移植组,尾静脉移植组高于创伤性脑损伤组(P<0.05)。提示移植骨髓间充质干细胞对创伤性脑损伤大鼠海马神经元有保护作用,脑损伤区移植优于尾静脉移植。
BACKGROUND： In recent years, application of bone marrow mesenchymal stem cells （BMSCs） transplantation in the central nervous system has become more widely, and the study of the best way to transplantation is important. OBJECTIVE： To investigate the effect of BMSCs transplantation in different ways on hippocanipal neuron following traumatic brain injury （TBI） in rats. METHODS： BMSCs were isolated from the bone marrow of 1-month-old SD rat. The BMSCs at passage 3 were labeled with PKH26 and were observed under fluorescence microscope. The percentage of the labeled cells was detected by flow cytometer. The model of hippocampal neuronal death following TBI was established by improved Feeney’s weight drop techniques. Sixty Wistar rats were divided into three groups randomly： Injured brain area transplantation group was injected with 10 μL normal saline contained BMSCs; Vena caudalis transplantation group was injected with 1 mL normal saline contained BMSCs through vein caudalis; TBI group was not treated. RESULTS AND CONCLUSION： Fluorescence microscope observation showed that the BMSCs labeled by PKH26 were spherical and presented with red fluorescence and uniform-distributed. Flow cytometric detection revealed that the percentage of labeled cells was over 99%. The learning and memory function of injured brain area transplantation group was significantly better than that of other two groups. The number of PKH26-positive cells and neurons after hematoxylin-eosin staining in injured brain area transplantation group was higher than that in vena caudalis transplantation group, and the number in vena caudalis transplantation group was higher that in TBI group （P 0.05）. RT-PCR approved that the expression of GAP-43 mRNA was highest in injured brain area transplantation group, a few in vena caudalis transplantation group and lowest in TBI group （P 0.05）. BMSCs transplantation plays an important role in protecting against hippocampal neuronal death following TBI. Injured brain area transplantation was significantly better than vena caudalis transplantation.
出处
《中国组织工程研究与临床康复》
CAS
CSCD
2012年第1期85-89,共5页
Journal of Clinical Rehabilitative Tissue Engineering Research
