定向诱导为神经元样细胞的骨髓间充质干细胞对大鼠坐骨神经损伤的修复作用研究  

Study on the repairing effect of BM-MSCs derived neuron-like cells on sciatic nerve injury in rats

作　　者：胡安全 冯祁军 王正安 秦力 李哲 HU Anquan;FENG Qijun;WANG Zheng’an;QIN Li;LI Zhe(Department of Orthopedics,Jiaxing First Hospital,Zhejiang Province,Jiaxing 314000,China)

机构地区：[1]浙江省嘉兴市第一医院骨科,浙江嘉兴314000

出　　处：《中国医药导报》2021年第6期27-33,F0003,共8页China Medical Herald

基　　金：浙江医药卫生科技计划项目(2016KYB291);浙江省嘉兴市科技计划项目(2017AY33009)。

摘　　要：目的研究定向诱导为神经元样细胞的骨髓间充质干细胞(BM-MSCs)对大鼠坐骨神经损伤的修复作用。方法取12只SD大鼠,分离纯化BM-MSCs后,将其分成对照组、β-巯基乙醇组、全反式视黄酸组、Transwell小室共培养组,分别诱导7 d,选择诱导分化率最高的细胞用于动物实验。另取90只大鼠,按随机数字表法分为假手术组、模型组、普通细胞组和定向诱导组,采用钳夹法建立大鼠坐骨神经挤压伤模型,最终每组各20只大鼠。普通细胞组和定向诱导组大鼠在坐骨神经损伤处分别注射BM-MSCs或神经元样定向诱导分化的BM-MSCs。假手术组和模型组注射等量生理盐水。比较四组坐骨神经功能指数(SFI)、Bsso Beattie Bresnahan(BBB)评分和肌湿重恢复率,Western blot法检测神经组织脑源性神经营养因子(BDNF)、髓磷脂碱性蛋白(MBP)和生长关联蛋白43(GAP-43)的表达水平。结果β-巯基乙醇用于后续动物实验。模型组术后2、4、8、12周SFI低于假手术组,差异均有统计学意义(均P<0.05)。定向诱导组术后2、4、8、12周SFI高于模型组、普通细胞组;普通细胞组术后2、4、8、12周SFI高于模型组,差异均有统计学意义(均P<0.05)。模型组术后2、4、8、12周BBB评分低于假手术组,差异均有统计学意义(均P<0.05)。定向诱导组术后8、12周BBB评分高于模型组和普通细胞组;普通细胞组术后8、12周BBB评分高于模型组,差异均有统计学意义(均P<0.05)。模型组大鼠肌湿重恢复率低于假手术组,差异有统计学意义(P<0.05)。定向诱导组肌湿重恢复率高于普通细胞组及模型组;普通细胞组肌湿重恢复率高于模型组,差异均有统计学意义(均P<0.05)。模型组BDNF、MBP与和GAP-43蛋白表达量低于假手术组,差异均有统计学意义(均P<0.05)。定向诱导组BDNF、MBP和GAP-43蛋白表达量高于模型组和普通细胞组;普通细胞组BDNF、MBP与和GAP-43蛋白表达量高于模型组,差异�Objective To investigate the repairing effect of bone marrow mesenchymal stem cells(BM-MSCs)derived neuron-like cells on sciatic nerve injury rats.Methods BM-MSCs from 12 SD rats were isolated and purified,and they were divided into control group,β-mercaptoethanol group,all-trans retinoic acid group and Transwell cell co-culture group for seven days respectively.The cells with the highest induced differentiation rate were selected for animal experiments.A total of 90 rats were divided into sham-operation group,model group,general cell group and directional induction group according to the random number table method.The sciatic nerve crush injury model was established by clamping method,and 20 rats in each group were selected.The rats of general cell group and directional induction group were injected with BM-MSCs or neuron-like BM-MSCs respectively at the site of injury sciatic nerve.The rats in sham-operation group and model group were injected with the same amount of normal saline.The sciatic nerve function index(SFI),Bsso Beattie Bresnahan(BBB)scores and the recovery rate of muscle wet weight were compared among four groups.The expression of brain-derived neurotrophic factor(BDNF),myelin basic protein (MBP) and growth associated protein 43 (GAP-43) of nerve tissue were detected by Western blot. Results β-mercaptoethanol was used in subsequent animal experiments. SFI of model group at 2, 4, 8 and 12 weeks after operation were lower than those of the sham-operation group, and the differences were statistically significant (all P < 0.05). SFI of directional induction group at 2, 4, 8 and 12 weeks after operation were higher than those of model group and general cell group;SFI of general cell group at 2, 4, 8 and 12 weeks after operation were higher than those of model group, and the differences were statistically significant (all P < 0.05). BBB scores in model group at 2, 4, 8 and 12 weeks after operation were lower than those in sham-operation group, and the differences were statistically significant (all P <

关 键 词：骨髓间充质干细胞 β-巯基乙醇 神经元样细胞 坐骨神经损伤 神经修复 

分 类 号：R688[医药卫生—骨科学]