Li Jun, Guo Weichun, Xiong Min, Zhang Shuangjie, Han Heng, Chen Jie, Mao Dan, Yu Hualong, Zeng Yun
Department of Orthopedics, Renmin Hospital of Wuhan University, Wuhan, Hubei 430060, P.R. China.
Department of Orthopedics, Dongfeng General Hospital of Hubei University of Medicine, Shiyan, Hubei 442008, P.R. China.
Exp Ther Med. 2017 May;13(5):1806-1812. doi: 10.3892/etm.2017.4182. Epub 2017 Mar 6.
Despite the successes of bone marrow mesenchymal stem cell (BMSC) transplantation for the treatment of spinal cord injuries, only a small fraction of grafted cells migrate to the target areas. Therefore, there remains a need for more efficient strategies of BMSC delivery. The present study was designed to explore this. Rat models of spinal cord injury (SCI) were established and exposed to phosphate buffered saline (control), BMSCs or BMSCs + erythropoietin (EPO). Basso, Beattie and Bresnahan (BBB) locomotor scale and grid walk tests were then utilized to estimate neurological rehabilitation. Additionally, the following assays were performed: Immunofluorescence localization of BMSCs to the site of SCI; the transwell migration assay to detect cellular migration; the terminal deoxynucleotidyl transferase dUTP nick end labeling assay to determine the apoptotic index of the lesion; and western blotting analysis to evaluate the expression of vascular endothelial growth factor (VEGF) and brain derived neurotrophic factor (BDNF) at the site of SCI. The BBB scores of the BMSC + EPO treated group were significantly increased compared with the BMSC treatment group (P<0.05). For example, BMSC + EPO treated rats had a significantly decreased number of hind limb slips compared with the BMSC treatment group (P<0.05). Furthermore, EPO significantly increased the migration capacity of BMSCs compared with the control group (P<0.001). In addition, the apoptotic index of the BMSC + EPO group was significantly decreased compared with the BMSC group (P<0.05). Green fluorescent protein-labeled BMSCs were detected at the site of SCI in the BMSC and BMSCs + EPO groups, with the signal being notably stronger in the latter. Moreover, the expression of VEGF and BDNF in the BMSCs + EPO group was significantly increased compared with the BMSC group (P<0.05). In conclusion, the results of the present study indicate that EPO can facilitate the recruitment of BMSCs to sites of SCI, increase expression of BDNF and VEGF, and accelerate recovery of neurological function following SCI.
尽管骨髓间充质干细胞(BMSC)移植治疗脊髓损伤取得了成功,但移植细胞中只有一小部分迁移到目标区域。因此,仍需要更有效的BMSC递送策略。本研究旨在探索这一问题。建立大鼠脊髓损伤(SCI)模型,并分别给予磷酸盐缓冲盐水(对照组)、BMSC或BMSC + 促红细胞生成素(EPO)处理。然后利用Basso、Beattie和Bresnahan(BBB)运动评分量表和网格行走试验评估神经功能恢复情况。此外,还进行了以下检测:BMSC在SCI部位的免疫荧光定位;检测细胞迁移的Transwell迁移试验;确定损伤部位凋亡指数的末端脱氧核苷酸转移酶dUTP缺口末端标记试验;以及评估SCI部位血管内皮生长因子(VEGF)和脑源性神经营养因子(BDNF)表达的蛋白质免疫印迹分析。与BMSC治疗组相比,BMSC + EPO治疗组的BBB评分显著提高(P<0.05)。例如,与BMSC治疗组相比,BMSC + EPO治疗的大鼠后肢滑倒次数显著减少(P<0.05)。此外,与对照组相比,EPO显著提高了BMSC的迁移能力(P<0.001)。另外,与BMSC组相比,BMSC + EPO组的凋亡指数显著降低(P<0.05)。在BMSC和BMSC + EPO组的SCI部位均检测到绿色荧光蛋白标记的BMSC,后者的信号明显更强。此外,与BMSC组相比,BMSC + EPO组中VEGF和BDNF的表达显著增加(P<0.05)。总之,本研究结果表明,EPO可促进BMSC募集至SCI部位,增加BDNF和VEGF的表达,并加速SCI后神经功能的恢复。