促红细胞生成素动员骨髓间充质干细胞治疗脊髓损伤

目的 观察促红细胞生成素(EPO)动员骨髓间充质干细胞(BMSCs)对脊髓损伤局部起的修复作用,并探讨其机制.方法 将30只SD大鼠随机分成假手术组、生理盐水组及EPO治疗组,各组均经尾静脉注射Hoechst33342标记的BMSCs,移植后1、3、7、14、21、28 d采用BBB法进行大鼠后肢运动功能评分观察大鼠神经功能恢复情况,应用免疫荧光检测比较各组BMSCs动员到脊髓损伤处的数目,Western blot检测损伤局部BMSCs表达脑源性神经营养因子(BDNF)和神经生长因子(NGF)的水平.结果　从第3天起,EPO治疗组BBB评分分别为(5.29±0.69)、(8.18±0.38)、(13.32±0.77)、(15.25±1.83)、(18.71±1.54),明显高于生理盐水组,移植的BMSCs计数分别为(90.12±7.68)、(116.26±13.54)、(186.32±20.35)、(211.64±31.83)、(298.71±21.54),数目明显增加(P＜0.05),其BDNF和NGF的表达也明显增强(P＜0.05).结论 EPO能够动员BMSCs向脊髓损伤部位迁移并通过促进BMSCs表达BDNF和NGF来促进神经功能的恢复.

Abstract：

Objective To obsevre the effect of erythropoietin (EPO) treatment on the mobility of bone marrow mesenchymal stem cell (BMSCs) to the sites of injured spinal cord and explore the molecular mechanism. Methods Thirty SD rats were randomly divided into sham-operation group, saline group and EPO treatment group. BMSCs labeled with Hoechst33342 were transplanted by tail vein. Functional outcome measurements were evaluated by the Basso, Beattie and Bresnahan (BBB) score at 1st, 3rd, 7th,14th, 21st and 28th day. The immigrating number of BMSCs was measured by immunofluorescent staining.The expression of brain-derived neurotrophic factor (BDNF) and nerve growth factor (NGF) in BMSCs was detected by Western blotting. Results From the 3rd day on, the BBB scores in EPO treatment group were (5.29±0.69), (8.18±0.38), (13.32±0.77), (15.25 ±1.83), (18.71 ±1.54) respectively, which were obviously higher than in saline group. The number of BMSCs was (90. 12 ±7. 68), (116. 26 ± 13. 54),( 186. 32 ± 20. 35 ), (211.64 ± 31.83 ) and (298.71 ± 21.54) respectively, which was significantly increased in EPO treatment group (P ＜ 0. 05 ). The protein levels of BDNF and NGF expressed by BMSCs were increased significantly (P ＜ 0. 05) in EPO treatment group as compared with sham-operation group.Conclusion EPO treatment after spinal cord injury may help BMSCs migrate to the injury site and accelerate recovery of neural function by up-regulating the expression of BDNF and NGF.

Effect of erythropoietin on the mobility of bone marrow mesenchymal stem cells to the sites of injured spinal cord

Objective To obsevre the effect of erythropoietin (EPO) treatment on the mobility of bone marrow mesenchymal stem cell (BMSCs) to the sites of injured spinal cord and explore the molecular mechanism. Methods Thirty SD rats were randomly divided into sham-operation group, saline group and EPO treatment group. BMSCs labeled with Hoechst33342 were transplanted by tail vein. Functional outcome measurements were evaluated by the Basso, Beattie and Bresnahan (BBB) score at 1st, 3rd, 7th,14th, 21st and 28th day. The immigrating number of BMSCs was measured by immunofluorescent staining.The expression of brain-derived neurotrophic factor (BDNF) and nerve growth factor (NGF) in BMSCs was detected by Western blotting. Results From the 3rd day on, the BBB scores in EPO treatment group were (5.29±0.69), (8.18±0.38), (13.32±0.77), (15.25 ±1.83), (18.71 ±1.54) respectively, which were obviously higher than in saline group. The number of BMSCs was (90. 12 ±7. 68), (116. 26 ± 13. 54),( 186. 32 ± 20. 35 ), (211.64 ± 31.83 ) and (298.71 ± 21.54) respectively, which was significantly increased in EPO treatment group (P ＜ 0. 05 ). The protein levels of BDNF and NGF expressed by BMSCs were increased significantly (P ＜ 0. 05) in EPO treatment group as compared with sham-operation group.Conclusion EPO treatment after spinal cord injury may help BMSCs migrate to the injury site and accelerate recovery of neural function by up-regulating the expression of BDNF and NGF.