减重步行训练对脊髓损伤鼠干细胞移植后神经细胞诱导分化的实验研究

发布时间：2018-03-16 10:43

  本文选题：脊髓损伤　切入点：干细胞移植　出处：《新疆医科大学》2015年硕士论文　论文类型：学位论文

【摘要】：目的：探讨骨髓间充质干细胞(Bone mesenchymal stemcells, BMSCs)移植联合减重步行训练对脊髓损伤(SCI)大鼠神经功能恢复的影响。方法：选取Sprague-Dawley(SD)雄性大鼠,采用改良的简易打击装置制作T11完全性SCI模型,最终将造模成功的40只SCI大鼠根据干预方法的不同按随机数字表法分为干细胞移植组、减重步行训练组、联合治疗组(干细胞移植联合减重步行训练)和对照组(不做任何干预处理),每组10只大鼠。脊髓损伤手术1周后,对干细胞移植组及联合治疗组大鼠进行干细胞移植。取传至第3代的BMSCs,移植前一天用5-乙炔基-2'脱氧尿嘧啶核苷(5-ethynyl-2'-deoxyuridine,Edu)标记,移植后减重步行训练组及联合治疗组进行减重步行训练,其余2组进行自由活动。SCI后第1、2、3、4和5周通过(Basso-Beattie-Bresnahan, BBB)运动功能评分来判断运动功能恢复情况,SCI第5周,应用免疫组化染色及免疫荧光染色的方法检测神经特异性标志物：神经元特异性烯醇化酶(NSE)、微管相关蛋白(MAP-1β)、波形蛋白(VIM)阳性表达,观察移植细胞的存活、分化及损伤部位神经纤维的恢复情况。结果：BBB运动功能评分显示,SCI手术第2周联合治疗组BBB评分为(6.60±0.97)分,明显高于其余3组,干细胞移植组和减重步行训练组BBB评分分别为(5.00±0.67)分、(4.80±0.63)分,均高于对照组,但两组之间BBB运动功能评分差异无统计学意义(P0.05)。术后第3周,干细胞移植组的BBB评分为(8.00±0.670)分,高于减重步行训练组BBB评分为(6.80±0.79)分,(P0.05)差异具有统计学意义。免疫组化染色法显示,有不同程度的神经特异性标志物(NSE、MAP-1β、VIM)阳性细胞充填于各组大鼠的SCI组织中,通过半定量分析法读取各组大鼠脊髓组织中阳性表达半定量值,联合治疗组分别为(4.72±0.19)分、(4.50±0.22)分、(4.62±0.27)分,明显高于其余3组(P0.05),组间差异具有统计学意义。免疫荧光染色法显示,联合治疗组神经特异性标志物(NSE、MAP-1β、VIM)荧光表达明显强于其余3组,同时可见明显的神经纤维增生、分化。结论：干细胞移植结合减重步行训练可有效促进SCI大鼠神经功能的恢复,效果明显优于单纯干细胞移植组和减重步行训练组。
[Abstract]:Objective: to investigate the effect of bone mesenchymal stem cells (BMSCs) transplantation combined with weight-loss walking training on the recovery of neural function in spinal cord injury (sci) rats. Finally, 40 successful SCI rats were randomly divided into stem cell transplantation group and weight-loss walking training group according to different intervention methods. The combined treatment group (stem cell transplantation combined with weight-loss walking training) and the control group (without any intervention, 10 rats in each group, 1 week after the operation of spinal cord injury), Stem cell transplantation was performed on rats in stem cell transplantation group and combined treatment group. BMSCs, which were transferred to the third generation, were labeled with 5-ethynyl-2deoxyuridine1, 5-ethynyl-2'deoxyuridine, 5-ethynyl-2'deoxyuridine1, and labeled with 5-ethynyl-2'deoxyuridine one day before transplantation. After transplantation, the weight loss walking training group and the combined treatment group were given weight-loss walking training. The other two groups were given free exercise. The recovery of motor function was evaluated by Basso-Beattie-Bresnahan (BBB) score at the 4th and 5th week after sci. Immunohistochemical staining and immunofluorescence staining were used to detect the positive expression of neuron-specific enolase (NSEN), microtubule-associated protein (MAP-1 尾) and vimentin (VIMM), and to observe the survival of transplanted cells. Results the BBB score of the combined treatment group was 6.60 卤0.97 at the 2nd week of sci, which was significantly higher than that of the other three groups. The BBB score of stem cell transplantation group and weight-loss walking training group was 4.80 卤0.63, which was higher than that of control group, but there was no significant difference in BBB motor function score between the two groups. The BBB score of stem cell transplantation group was 8.00 卤0.670 at the third week after operation. The BBB score was 6.80 卤0.79 (P 0.05) higher than that in the weight-loss walking training group (P 0.05). Immunohistochemical staining showed that there were different degrees of NSEMAP-1 尾 -VIM-positive cells filled in the SCI tissues of rats in each group. The semi-quantitative analysis method was used to read the semi-quantitative value of positive expression in spinal cord tissue of rats in each group. The score of positive expression in the combined treatment group was 4.72 卤0.19) and 4.50 卤0.22), which was significantly higher than that in the other three groups (P 0.05), and the difference between the two groups was statistically significant. The fluorescence expression of NSEP MAP-1 尾 -VIMM in the combined treatment group was significantly higher than that in the other three groups, and obvious nerve fiber proliferation and differentiation could be observed. Conclusion: stem cell transplantation combined with weight-loss walking training can effectively promote the recovery of nerve function in SCI rats. The effect was better than that of stem cell transplantation group and walking training group.
【学位授予单位】：新疆医科大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R651.2
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本文编号：1619574