论文部分内容阅读
目的:在制作新生大鼠缺氧缺血性脑损伤模型基础上,研究间质干细胞移植治疗缺氧缺血性脑病的应用前景。方法:实验于2002-09/2004-03在深圳宝安血站干细胞研究实验室内完成。实验中采用L-DMEM分离培养骨髓中间质干细胞。7窝SD1d龄新生大鼠共81只同窝随机分为4组:①正常组,不干预。②模型组:结扎左颈动脉,吸氧3h制成缺氧缺血性模型。③生理盐水组:造模后24h定位注射生理盐水8μL。④间质干细胞组:处理同生理盐水组,注射间质干细胞(1.0～2.0)×106/μL。Morris水迷宫试验评估移植后第42天大鼠学习和记忆能力;第10周麻醉状态下处死实验鼠,制作病理切片,苏木精-伊红染色和间接免疫荧光检测间质干细胞在脑内存活和分化情况。结果:①第1周内成活率:第7天,81只实验鼠存活42只,间质干细胞组与正常对照组大鼠存活率分别为75.0%和82.4%,差异不显著;明显高于生理盐水组(35.3%)和模型组(50.0%)(P<0.01)。②水迷宫试验显示间质干细胞组大鼠的空间识别和记忆能力明显优于生理盐水组和模型组(P<0.01);培训后第5天,间质干细胞组识别平台的平均时间为7.5s,与正常组7.0s不存在差异。③病理切片、苏木精-伊红染色和间接免疫实验结果显示:间质干细胞移植组中,进针注射部位存在大量移植细胞,并向周围迁移,皮质层、海马等部位检测到移植细胞;所植入的细胞约6%表达胶质纤维酸性蛋白,约8%表达神经元特异性烯醇化酶。结论:移植人间质干细胞组能有效修复缺氧缺血导致的神经受损,改善其功能,明显提高缺氧缺血性脑损伤大鼠1周内成活率。
Objective: To study the application prospect of mesenchymal stem cell transplantation in the treatment of hypoxic-ischemic encephalopathy based on the model of neonatal rats with hypoxic-ischemic brain damage. Methods: The experiment was completed in Shenzhen Baoan Blood Station Stem Cell Research Laboratory from September 2002 to March 2004. In the experiment, bone marrow mesenchymal stem cells were isolated and cultured by L-DMEM. A total of 81 neonate SD1d neonate rats were randomly divided into 4 groups: ① normal group without intervention. ② model group: ligation of the left carotid artery, oxygen 3h hypoxia-ischemic model. ③ saline group: 24h after injection modeling saline injection 8μL. ④ Mesenchymal stem cells group: treated with saline, injected mesenchymal stem cells (1.0 ~ 2.0) × 106 / μL. The Morris water maze test was used to assess the learning and memory abilities of rats on the 42nd day after transplantation. At week 10, the rats were sacrificed and anesthetized, pathological sections were made, hematoxylin-eosin staining and indirect immunofluorescence were used to detect the survival of mesenchymal stem cells And differentiation. Results: ① The survival rate in the first week: on the seventh day, the survival rate of 81 rats was 42, the survival rates of the MSCs group and the normal control group were 75.0% and 82.4% respectively, with no significant difference; Saline group (35.3%) and model group (50.0%) (P <0.01). ② The water maze test showed that the spatial recognition and memory ability of mesenchymal stem cell group was significantly better than that of saline group and model group (P <0.01). On the fifth day after training, the mean time of recognition platform of mesenchymal stem cell group was 7.5s , There is no difference with the normal group 7.0s. ③ Pathological sections, hematoxylin-eosin staining and indirect immunization results showed that a large number of transplanted cells were transplanted to the injection site of MSCs, and the transplanted cells were detected in cortex and hippocampus. About 6% of the implanted cells express glial fibrillary acidic protein and about 8% express neuron-specific enolase. Conclusion: Transplantation of human mesenchymal stem cells can effectively repair the damage caused by hypoxia-ischemia and improve its function, and significantly improve the survival rate of rats in hypoxic-ischemic brain damage within one week.