南方医科大学学报

南方医科大学学报 ›› 2019, Vol. 39 ›› Issue (06): 685-.doi: 10.12122/j.issn.1673-4254.2019.06.09

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大鼠脑损伤运动障碍模型中移植脂肪干细胞来源的施旺细胞后细胞缝隙连接在神经功能修复中的作用

杨幼萌，杨靓，王知非   

  • 出版日期:2019-06-20 发布日期:2019-06-20

Formation of gap junctions between adipose stem cells-derived Schwann cells in a rat model of dyskinesia induced by brain injury

  • Online:2019-06-20 Published:2019-06-20

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摘要： 目的证明移植脂肪干细胞及其诱导分化而来的施旺细胞有利于急性脑损伤动物模型的功能恢复。探索细胞移植后缝隙 连接在脑损伤修复过程中的作用机制。方法在运动皮层损伤偏瘫大鼠模型中，将培育分化得到的脂肪干细胞及施旺细胞以及 RNAi技术沉默Cx43基因的施旺细胞移植损伤灶处行原位培养，术后观察大鼠运动功能恢复情况并予评分。移植7 d后处死取 鼠脑组织，RT-PCR和Western blot验证神经生长因子（NGF）的表达水平。结果在建立的运动皮层损伤的偏瘫大鼠模型，细胞 移植组大鼠优于对照组运动评分。WB结果中对照组NGF表达明显低于各移植组，Cx43基因沉默的施旺细胞移植后组织NGF 表达低于未沉默组，Cx43基因未沉默组与脂肪干细胞移植组无显著性差异。RT-PCR法测得对照组NGF mRNA水平明显低于 各移植组，施旺细胞组水平高于脂肪干细胞组、高于Cx43基因沉默施旺细胞组。结论移植脂肪干细胞和其经诱导分化而来的 施旺细胞，都有利于动物模型的脑损伤后功能恢复。在脑损伤修复期功能性缝隙连接形成过程中，缝隙连接蛋白Cx43发挥了 重要作用，可能通过促进神经生长因子NGF的表达有关。

Abstract: Objective To investigate the formation of gap junctions between Schwann cells derived from differentiated adipose stem cells implanted in a rat model of dyskinesia induced by brain injury and its positive effect in promoting functional recovery of the rats. Methods In a rat model of hemiplegia induced by motor cortex injury, adipose stem cells or Schwann cells differentiated from adipose stem cells, either with or without RNAi-mediated silencing of Cx43, were transplanted orthotopically in the lesion. The recovery of the motor function of the rats was observed and scored after the transplantation. Rat brain tissues were sampled to detect the expressions of nerve growth factor (NGF) using Western blotting and RT-PCR. Results All the 3 cell transplantation therapies obviously improved the motor function scores of the rats as compared with the control rats. The expression of NGF in the brain tissue was significantly lower in the control group than in the cell transplantation groups. NGF expression in the brain tissues of rats receiving transplantation of Schwann cells with Cx43 gene silencing was lower than that in rats receiving Schwann cells without Cx43 silencing, and was similar with that in rats transplanted with adipose stem cells. The results of RT-PCR showed that NGF mRNA level in the control group was significantly lower than that in the other 3 groups. NGF mRNA expression was the highest in Schwann cell group without Cx43 silencing, followed by adipose stem cell group, and then by Schwann cell group with Cx43 silencing. Conclusion In the rat model of dyskinesia induced by brain injury, transplantations of adipose stem cells and adipose stem cells-derived Schwann cells both promote the functional recovery of brain damage, in which gap junction protein Cx43 plays an important role to promote functional gap junction formation possibly by enhancing NGF expression.