Gou Yang, Liu Danyan, Liu Jinfeng, Sun Hongran
Department of Anesthesiology, the First Affiliated Hospital of Chongqing Medical University, Chongqing 400042, China.
Sheng Wu Gong Cheng Xue Bao. 2018 May 25;34(5):761-776. doi: 10.13345/j.cjb.170406.
Bone mesenchymal stem cells (BMSCs) have been used worldwide to treat spinal cord injury, but their therapeutic mechanism is poorly understood. In this study, BMSCs were transplanted to aneurysm clip-injured rats to demonstrate their protective effect. We observed myelin sheaths through Luxol fast blue (LFB) staining, osmic acid staining, TUNEL and transmission electron microscopy (TEM). We performed Western blotting to analyze the expressions of brain-derived neurotrophic factor (BDNF) and caspase 3. BMSCs were transplanted at 1, 7 and 14 days after spinal cord injury. Hindlimb movement (Basso, Beattie and Bresnahan; BBB) score, CNPase (2', 3'-cyclic-nucleotide 3'-phosphodiesterase), myelin basic protein (MBP) and caspase 3 protein levels were detected. Immunofluorescence was used to test the differentiation of BMSCs after implanted into damaged spinal cord and co-expression of CNPase-caspase 3+. At 7 days after BMSCs transplantation, some injected BMSCs expressed neuronal and oligodendrocyte markers. And both locomotor skills and ultra-structural features of myelin sheaths were significantly improved. The expressions of BDNF were clearly increased by BMSCs transplantation, the expression of caspase 3 was the opposite. Compared with the 1 and 14 days transplantation after spinal cord injury, MBP and CNPase expressions were highest, caspase 3 expression was lowest in 7 days BMSCs transplantation. After BMSCs transplantation, CNPase-caspase 3+ cells scattered in the white matter of the spinal cord. Therefore, BMSCs had a tendency to differentiate into neurons and oligodendrocytes after transplantation, which could promote the secretion of BDNF. BMSCs protected neural myelin sheaths by inhibiting oligodendrocyte apoptosis via increased secretion of BDNF after SCI. The best therapeutic time was 7 days after spinal cord injury.
骨髓间充质干细胞(BMSCs)已在全球范围内用于治疗脊髓损伤,但其治疗机制尚不清楚。在本研究中,将BMSCs移植到动脉瘤夹损伤的大鼠中以证明其保护作用。我们通过Luxol固蓝(LFB)染色、锇酸染色、TUNEL和透射电子显微镜(TEM)观察髓鞘。我们进行蛋白质免疫印迹分析脑源性神经营养因子(BDNF)和半胱天冬酶3的表达。在脊髓损伤后1、7和14天移植BMSCs。检测后肢运动(Basso、Beattie和Bresnahan;BBB)评分、2',3'-环核苷酸3'-磷酸二酯酶(CNPase)、髓鞘碱性蛋白(MBP)和半胱天冬酶3蛋白水平。免疫荧光用于检测植入受损脊髓后BMSCs的分化以及CNPase-半胱天冬酶3+的共表达。在BMSCs移植后7天,一些注入的BMSCs表达神经元和少突胶质细胞标志物。并且运动技能和髓鞘的超微结构特征均得到显著改善。BMSCs移植明显增加了BDNF的表达,而半胱天冬酶3的表达则相反。与脊髓损伤后1天和14天移植相比,在BMSCs移植7天时MBP和CNPase表达最高,半胱天冬酶3表达最低。BMSCs移植后,CNPase-半胱天冬酶3+细胞散在于脊髓白质中。因此,BMSCs移植后有分化为神经元和少突胶质细胞的趋势,这可以促进BDNF的分泌。脊髓损伤后,BMSCs通过增加BDNF的分泌抑制少突胶质细胞凋亡,从而保护神经髓鞘。最佳治疗时间为脊髓损伤后7天。