舒芬太尼对神经干细胞移植治疗脊髓损伤的影响

发布时间：2018-03-11 08:12

本文选题：脊髓损伤　切入点：神经干细胞　出处：《中国组织工程研究》2017年25期 　论文类型：期刊论文

【摘要】：背景:有研究显示舒芬太尼联合神经干细胞移植联合治疗可以提高新生的神经纤维数量。目的:观察舒芬太尼干预对神经干细胞移植治疗脊髓损伤大鼠后肢功能恢复的影响。方法:(1)将100只成年雌性SD大鼠随机分为5组:正常对照组、模型组、舒芬太尼组、神经干细胞移植组和舒芬太尼+神经干细胞移植联合治疗组,每组20只。正常对照组大鼠不实施任何干预,其他4组参照改良的Allen’s重物打击法建立大鼠脊髓损伤模型;(2)造模6 h后,联合治疗组大鼠蛛网膜下腔注射10μL细胞浓度为1×10~(10) L~(-1)的神经干细胞悬液,腹腔注射100μL舒芬太尼(150μg/kg);神经干细胞移植组大鼠蛛网膜下腔注射10μL细胞浓度为1×10~(10) L~(-1)的神经干细胞悬液,腹腔注射100μL生理盐水;舒芬太尼组蛛网膜下腔注射10μL的神经干细胞培养液,腹腔注射100μL舒芬太尼(150μg/kg);模型组大鼠蛛网膜下腔和腹腔各注射10μL神经干细胞培养液和100μL生理盐水;(3)造模后72 h,RT-PCR检测脊髓损伤区周围AQP4、MMP9 mRNA的表达,TUNEL法检测细胞凋亡情况;(4)造模后第1,3天和第1,2,3,4周通过BBB评分、斜板实验进行运动功能评定;(5)造模后4周取材行苏木精-伊红染色,荧光显微镜观测CM-Dil标记的神经干细胞存活情况,荧光金逆行追踪观察脊髓神经纤维的再生与分布情况。结果与结论:(1)造模后72 h,与模型组、舒芬太尼组、神经干细胞移植组比较,联合治疗组脊髓损伤区AQP4、MMP9 mRNA表达和细胞凋亡率显著降低(P0.05);(2)造模2周后,联合治疗组运动功能评分优于舒芬太尼组和神经干细胞移植组(P0.05),舒芬太尼组和神经干细胞移植组优于模型组(P0.05);(3)造模后4周,模型组可见脊髓空洞形成,舒芬太尼组和神经干细胞移植组损伤区脊髓空洞较小,联合治疗组脊髓空洞几乎消失;(4)联合治疗组CM-Dil阳性细胞最多,神经干细胞移植组较多,舒芬太尼组和模型组未见阳性细胞;(5)联合治疗组荧光金阳性神经纤维数最多,舒芬太尼组和神经干细胞移植组次之,模型组最少;(6)结果表明,舒芬太尼通过促进移植神经干细胞的增殖,减少大鼠脊髓损伤区AQP4、MMP9表达和局部神经细胞凋亡,进而改善后肢运动功能。
[Abstract]:Background: studies have shown that sufentanil combined with neural stem cell transplantation can increase the number of newborn nerve fibers. Objective: to observe the effects of sufentanil on the function of hindlimb of spinal cord injury rats treated by neural stem cell transplantation. Methods 100 adult female SD rats were randomly divided into 5 groups: normal control group. Model group, sufentanil group, neural stem cell transplantation group and sufentanil neural stem cell transplantation combined treatment group, 20 rats in each group. In the other four groups, the spinal cord injury model of rats was established by using the modified Allen's weight attack method for 6 h. The rats in the combined treatment group were injected into subarachnoid space with 10 渭 L cell concentration of 1 脳 10 ~ (10) L ~ (-1) L ~ (-1) neural stem cell suspension. 100 渭 L sufentanil 150 渭 g 路kg ~ (-1) was injected intraperitoneally into 100 渭 L sufentanil 150 渭 g 路kg ~ (-1), 10 渭 L neural stem cell suspension (10 渭 L cell concentration was 1 脳 10 ~ (10)) L ~ (-1) in the neural stem cell transplantation group, 100 渭 L normal saline was injected intraperitoneally, and 10 渭 L neural stem cell culture medium was injected into the subarachnoid space of sufentanil group. 100 渭 L sufentanil 150 渭 g / kg; subarachnoid and intraperitoneal injection of 10 渭 L neural stem cell culture medium and 100 渭 L normal saline in model group) 72 h later, RT-PCR was used to detect the expression of AQP4MMP9 mRNA around spinal cord injury area and Tunel method was used to detect apoptosis. The BBB score was obtained on the 1st 3rd day and 4th week of the 1st day and the 2nd week after the model was made. Four weeks after the model was made, the rats were stained with hematoxylin and eosin, and the survival of neural stem cells labeled with CM-Dil was observed by fluorescence microscope. The regeneration and distribution of nerve fibers in spinal cord were observed by fluorescence gold retrograde tracing. Results and conclusion the results were compared with those of model group, sufentanil group and neural stem cell transplantation group at 72 h after the establishment of the model group, sufentanil group and neural stem cell transplantation group. In the combined treatment group, the expression of MMP9 mRNA and the apoptotic rate of AQP4mMP9 in spinal cord injury area were significantly decreased 2 weeks after the establishment of the model. The motor function score in the combined treatment group was better than that in the sufentanil group and the neural stem cell transplantation group (P 0.05), and in the sufentanil group and the neural stem cell transplantation group, the spinal cord syringomyelia was found in the model group 4 weeks after the establishment of the model. In sufentanil group and neural stem cell transplantation group, syringomyelia in the injured area was smaller, and syringomyelia almost disappeared in the combined treatment group. The number of CM-Dil positive cells in the combined treatment group was more than that in the neural stem cell transplantation group. No positive cells were found in Sufentanil group and model group. The number of fluorescent gold positive nerve fibers was the highest in combined treatment group, followed by sufentanil group and neural stem cell transplantation group. Sufentanil could improve the motor function of hindlimb by promoting the proliferation of transplanted neural stem cells, reducing the expression of AQP4MMP9 and local neuronal apoptosis in spinal cord injury area of rats.
【作者单位】：滨州医学院附属医院麻醉科;滨州医学院附属医院药剂科;
【分类号】：R651.2

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