摘要
目的 探讨骨髓间充质干细胞(BMSCs)靶向移植和促红细胞生成素(EPO)联合对脊髓损伤的修复作用.方法 采用纽约大学(NYU)脊髓损伤模型打击器建立建立大鼠脊髓损伤模型,60只大鼠随机分为5组,每组12只,分别为对照组、模型组、BMSCs组、EPO组、BMSCs+ EPO组.BMSCs组在在模型组基础上向脊髓损伤处注射10μl(5×106个)BMSCs;EPO组连续5d腹腔注射5 IU/kg EPO;BMSCs+ EPO组在BMSCs组的基础上连续5d腹腔注射5 IU/kg EPO.采用Basso-Beattie-Bresnahan (BBB)评分标准评价大鼠后肢运动功能恢复;术后第0、8周后采用苏木素-伊红(HE)染色观察脊髓组织形态变化;采用Western blot检测脊髓损伤组织中神经生长因子(NGF)、神经营养素-3(NT-3)、脑源性神经营养因子(BDNF)、胶质细胞源性神经营养因子(GDNF)、睫状神经营养因子(CNTF)的表达水平.结果 与对照组BBB评分[(20.8±0.4)分]比较,模型组[(6.9±1.3)分]、BMSCs组[(12.9±2.1)分]、EPO组[(14.2±1.9)分]和BMSCs+EPO组[(17.9±2.8)分]均减少,差异有统计学意义(P<0.05);与模型组比较,BMSCs组、EPO组和BMSCs+ EPO组BBB评分增加,差异有统计学意义(P<0.05);与BMSCs组、EPO组比较,BMSCs+ EPO组BBB评分增加,差异有统计学意义(P<0.05).与对照组比较,模型组、BMSCs组、EPO组和BMSCs+ EPO组NGF、NT-3、BDNF、GDNF、CNTF表达水平增加,差异有统计学意义(P<0.05);与模型组比较,BMSCs组、EPO组和BMSCs+ EPO组NGF、NT-3、BDNF、GDNF、CNTF表达水平增加,差异有统计学意义(P<0.05);与BMSCs组和EPO组比较,BMSCs+ EPO组NGF、NT-3、BDNF、GDNF、CNTF表达水平增加,差异有统计学意义(P<0.05).HE染色结果显示,BMSCs组、EPO组和BMSCs+ EPO组脊髓组织都得到较好修复,BMSCs+ EPO组效果最好.结论 采用BMSCs靶向移植和EPO联合治疗脊髓损伤可以有效地促进脊髓损伤大鼠脊髓损伤再生和修复.
Objective To explore the effects about combined treatment of targeted bone marrow mesenchymal stem cells (BMSCs) transplantation and erythropoietin (EPO) in spinal cord injury.Methods New York University (NUY) method was adopted to establish the rat model of spinal cord injury.Sixty rats were randomly divided into 5 groups:control group,model group,BMSCs group,EPO group,and BMSCs + EPO group (n =12 each).Rats in BMSCs group were injected with 10 μl (5 × 106) BMSCs in spinal cord lesions after spinal cord injury.Rats in EPO group were intraperitonealy injected with 5 IU/kg EPO for 5 consecutive days.Rats in BMSCs + EPO group were treated with both BMSCs and EPO.The Basso-Beattie-Bresnahan (BBB) score was used to evaluate motor function recovery of hind limbs.Hematoxylin and eosin (HE) staining was used to evaluate morphological changes of spinal cord tissue 0,8 weeks after operation.Western blotting was used to detect the expression of nerve growth factor (NGF),nerve nutrients-3 (NT-3),brain derived neurotrophic factor (BDNF),glial cell derived neurotrophic factor (GDNF),and ciliary neurotrophic factor (CNTF) in spinal cord 0,8 weeks after injury.Results As compared with control group (20.8 ± 0.4),BBB score in model group (6.9 ± 1.3),B MSCs group (12.9±2.1),EPO group (14.2±1.9) and BMSCs + EPO group (17.9±2.8) decreased significantly (P 〈 0.05).As compared with model group,BBB score in BMSCs group,EPO group and BMSCs + EPO group increased significantly (P 〈 0.05).As compared with BMSCs group and EPO group,BBB score in BMSCs + EPO group increased significantly (P 〈0.05).As compared with control group,there were significant differences in the expression of NGF,NT-3,BDNF,GDNF,and CNTF in model group,BMSCs group,EPO group and BMSCs + EPO group 8 weeks after surgery (P 〈 0.05).As compared with model group,there were significant differences in the expression of NGF,NT-3,BDNF,GDNF,and CNTF in BMSCs group,EPO goup and BMSCs + EPO group 8 weeks after surgery (P 〈 0.05).As com-pared with BMSCs group and EPO group,there were significant differences in the expression of NGF,NT-3,BDNF,GDNF,and CNTF in BMSCs + EPO group 8 weeks after surgery (P 〈 0.05).HE staining revealed that spines in BMSCs group,EPO group and BMSCs + EPO group were better repaired,among which BMSCs + EPO group was the best.Conclusion The targeted BMSCs transplantion combined with EPO therapy can effectively promote regeneration and repair of spinal cord injury in rats.
作者
郑竑
张韬
林振恩
涂少臣
陈学生
Zheng Hong Zhang Tao Lin Zhen'en Tu Shaochen Chen Xuesheng(Department of Orthopedics, Fuzhou Second Hospital Affiliated to Xiamen University, Fuzhou 350007, Chin)
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2016年第11期2490-2492,共3页
Chinese Journal of Experimental Surgery
基金
2015年福州市科技计划项目(2015-S-141-3)
2015年福州市临床重点专科建设项目(骨科)(2015003)