人羊膜间充质干细胞移植对大鼠脊髓损伤后NGF、BDNF和Nogo-A mRNA表达的影响

Effects of human amnion-derived mesenchymal stem cells transplantation on the expression of NGF mRNA, BDNF mRNA, Nogo-A mRNA in rats after spinal cord injury

中文关键词:  脊髓损伤  移植  人羊膜间充质干细胞  神经营养因子  大鼠

英文关键词:Spinal cord injury  Transplantation  Human amnion-derived mesenchymal stem cell  Neurotrophic factor  Rats

中文摘要:

  【摘要】 目的：观察人羊膜间充质干细胞（hAMSCs）移植对大鼠脊髓损伤后神经生长因子（NGF）、脑源性神经营养因子（BDNF）及轴突生长抑制因子（Nogo-A）mRNA表达的影响。方法：成年雌性SD大鼠24只，随机分成hAMSCs移植组（12只）、磷酸盐缓冲液（PBS）对照组（12只）。体外分离培养、鉴定hAMSCs，并以终浓度为10μg/ml 4，6-二脒基-2-苯基吲哚（DAPI）荧光染料标记hAMSCs。在建立大鼠T11脊髓全横断损伤模型后，立即以微量注射器吸取约3×106个hAMSCs悬液于横断损伤处头尾两端原位移植，对照组注射相同体积的PBS液。分别于术后3d、7d利用荧光显微镜观察hAMSCs存活情况，并采用RT-PCR分析损伤脊髓段NGF mRNA、BDNF mRNA及Nogo-A mRNA的表达。结果：术后3d和7d hAMSCs移植组均可观察到植入的hAMSCs在宿主脊髓组织内存活。术后3d、7d，hAMSCs移植组NGF mRNA的表达均高于对照组（P＜0.05，P＜0.01），Nogo-A mRNA的表达均低于对照组（P＜0.05）；术后3d时hAMSCs移植组BDNF mRNA的表达与对照组比较无显著性差异（P＞0.05），但术后7d其表达高于对照组（P＜0.05）。结论：hAMSCs移植可上调大鼠受损脊髓组织内NGF mRNA和BDNF mRNA的表达、下调Nogo-A mRNA的表达，有利于促进脊髓损伤后的功能修复。

英文摘要:

  【Abstract】 Objectives: To observe the effects of human amniotic mesenchymal stem cells(hAMSCs) transplantation on the expression of nerve growth factor(NGF), brain-derived neurotrophic factor(BDNF) and axonal growth inhibitor(Nogo-A) mRNA in rats after spinal cord injury. Methods: Twenty-four adult female SD rats were randomly divided into transplantation group and control group. Transected spinal cord injury(SCI) model in rats was established. Isolation, culture and identification of hAMSCs in vitro were performed, and 10μg/ml DAPI was used to label cultured hAMSCs. About 3×106 hAMSCs absorbed by a microsyringe were transplanted into the head and tail end lesions immediately after establishing of animal model, the control group was injected by equal volume of PBS. The survival hAMSCs in the injured spinal cord were observed by fluorescence microscope at 3, 7 days post-operation. Meanwhile, the expression of NGF mRNA, BDNF mRNA and Nogo-A mRNA in the injured spinal cord was detected by RT-PCR. Results: The grafting hAMSCs were observed in the host spinal cord tissue at 3d and 7d post operation. RT-PCR results showed the expression of NGF mRNA in transplantation group was significantly higher than that in control group(P＜0.05, P＜0.01) regardless of timepoint, and the expression of Nogo-A mRNA was lower than that in the control group(P＜0.05). However, compared with the control group, the expression of BDNF mRNA at 3d after operation showed no significant difference(P＞0.05), but at 7d after operation, the expression of BDNF mRNA in transplantation group was remarkably higher than that of control group(P＜0.05). Conclusions: The transplantation of hAMSCs can regulate the expression of neurotrophic factors in injured spinal cord tissue, which may contribute to the improvement of neurological function of rats after SCI.