BMSC对大鼠脊髓损伤神经凋亡的保护作用及相关机制研究

摘要

Objective To explore the effects of bone mesenchymal stem cells (BMSC)on the spinal cord injury and the potential mecha-nisms. Methods BMSC were obtained via adherence method,and the cell phenotype was determined by flow cytometry. Spinal cord injury model was induced by using the modified weight - drop method at T8 level. 42 SD rats were randomly assigned to control group,SCI group and BMSC +SCI group. The rat hind limb was determined by the BBB(Basso,Beattie,Bresnahan,BBB)at the 7 d,14 d and 21 d. At the day of 21,all groups of spinal cord were taken out and the HE staining and immunohistochemistry were valued. The protein expression of Bax and Bcl - 2 were detected by western blot and the content of caspase - 3 were valued by elisa. Results The CD90 expression of BMSC were (98. 92 ± 0. 62)%, while the CD34 were (0. 47 ± 0. 16)% . BMSC could effectively improve the BBB scores of the spinal cord injury (P < 0. 05). Immunohisto-chemistry showed that BMSC could markedly suppress the content of Bax and caspase - 3 (P < 0. 05). Western blot showed that BMSC could sig-nificantly suppress Bax and enhance Bcl - 2 protein expression (P < 0. 05). Elisa showed that BMSC could markedly down - regulate the content of caspase - 3 (P < 0. 05). Conclusion The anti - neural apoptosis effects of BMSC on the spinal cord injury are via suppressing the Bax and caspase - 3 protein expression and improving the Bcl - 2 pathway.%目的 探索骨髓间充质干细胞(BMSC)对大鼠脊髓损伤(SCI)后神经凋亡的作用及其相关机制.方法体外获取大鼠BMSC进行增殖培养,利用流式细胞仪检测BMSC细胞表型,Allen法制作大鼠脊髓损伤模型,42只大鼠随机等分为3组,对照组,SCI组和BMSC+SCI组,分别于7 d、14 d和21 d对各组大鼠下肢运动功能评分;21 d后取出各组大鼠损伤脊髓进行组织切片染色,并利用Western blot法检测脊髓Bax/Bcl-2含量,ELISA法检测Caspase-3含量.结果BMSC细胞表型CD90表达(98.92±0.62)%,CD34表达(0.47±0.16)%;BMSC在7 d、14 d和21 d可明显改善SCI大鼠下肢运动功能(P均<0.05);免疫组化结果显示,BMSC可显著抑制Bax和Caspase-3在损伤脊髓中的表达(P均<0.05);Western blot显示,BMSC明显抑制Bax,提高Bcl-2表达(P均<0.05);ELISA结果显示,BMSC可显著机制损伤脊髓中Caspase-3含量(P均<0.05).结论BMSC对大鼠脊髓损伤神经凋亡具有明显抑制作用,可能是通过抑制Bax、Caspase-3,提高Bcl-2蛋白表达实现的.