脱细胞异种神经桥接体联合经血源性间充质干细胞对大鼠坐骨神经缺损的影响  

Effects of acellular nerve xenografts bridging combined with menstrual blood mesenchymal stem cells on sciatic nerve defects in rats

作　　者：王红燕 王贺颖[1] 梁雪云 贾桦[1] 段昉[4] 张博闻 仇莽莽[1] 何仲义 Wang Hongyan;Wang Heying;Liang Xueyun;Jia Hua;Duan Fang;Zhang Bowen;Qiu Mangmang;He Zhongyi(Depatment of Human Anatomy and Histology Embryology,Ningxia Medical University,Yinchuan 750004,China;Spem Bank of General Hospital,Ningxia Medical University,Yinchuan 750004,China;Ningxia Human Stem Cell Research Insituion of General Hospital,Ningxia Medical University,Yinchuan 750004,China;Yinchuan Women and Child Healtheare Hopital,Ningxia Medical University,Yinchuan 750004,China;People's Hospital of Ningpia Hui Autonomous Region,Ningxia Medical University,Yinchuan 750004,China)

机构地区：[1]宁夏医科大学基础医学院人体解剂与组织胚胎学系,银川750004 [2]宁夏医科大学总医院精子库,银川750004 [3]宁夏医科大学总医院干细胞研究所,银川750004 [4]宁夏医科大学附属银川市妇幼保健医院,银川750004 [5]宁夏医科大学附属自治区人民医院,银川750004

出　　处：《神经解剖学杂志》2020年第2期175-181,共7页Chinese Journal of Neuroanatomy

基　　金：宁夏自然科学基金项目(NZ2018AAC03066);宁夏重点研发基金项目(2019BEG03002)。

摘　　要：目的:研究脱细胞异种神经桥接体(ANX)联合经血源性间充质干细胞(MenSCs)对大鼠坐骨神经缺损的影响。方法:将SD大鼠随机分为自体移植组、单纯ANX组和ANX+MenSCs共培养组,制备SD大鼠右侧坐骨神经缺损模型,将不同组的桥接体移植到SD大鼠右侧坐骨神经缺损处,移植术后8周,用热板实验检测大鼠感觉功能的恢复,斜坡实验来检测运动功能的恢复,通过坐骨神经功能指数(SFI)检测大鼠坐骨神经功能,通过胫骨前肌湿重比率检测神经肌肉的功能,用透射电镜来观察髓鞘的恢复情况。结果:扫描电镜显示ANX组内原有的细胞及结构基本去除,完整保留了神经的基膜及外膜,ANX+MenSCs组可见圆形的细胞在基膜间稳定黏附生长;免疫荧光染色显示神经生长因子(NGF)在ANX+MenSCs共培养组中有阳性表达;移植术后8周,ANX+MenSCs组的患肢回缩时间显著短于单纯ANX组(P<0.05),爬坡坡度、SFI值以及胫骨前肌湿重比率均显著高于单纯ANX组(P<0.05),ANX+MenSCs组与自体组相比,两组的患肢回缩时间、爬坡坡度以及胫骨前肌湿重比均无显著差异(P>0.05),但自体组的SFI值显著增高(P<0.05);透射电镜显示与单纯ANX组相比,ANX+MenSCs组和自体组的髓鞘结构更完整,虽有水肿,但形状小而均匀致密。结论:ANX联合MenSCs修复SD大鼠坐骨神经缺损的效果良好,MenSCs可能成为外周神经系统疾病细胞治疗的潜在候选种子细胞。Objective:To study the eecs of acellular nerve xenografis(ANX)combined with menstrual blood mes-enchymal stem ells(MenSCs)on sciatic nerve defect in SD rats.Methods:The SD rats were randomly divided into autograft group,ANX and ANX+MenSCs co-culture group.And then the SD rats right sciatie nerve defect models were prepared,and bridges of different groups were transplanted to the defects of the right sciatic nerve in SD rats.In eight weeks after the operation,the sensory function of the SD rats was measured by a hot plate experiment,the motor func-tion was measured by a ramp experiment,while the recovery of sciatic nerve functions was evaluated by sciatie nerve funcion index(SFI),and the neuromuscular function was measured by the tibialis anterior muscle wet weight ratio function,we use transmision electron microscope to observe the recovery of myelin sheath.Results:Scanning electron microscopy showed that the original cells and structures in ANX group were almost removed,with only the basal mem-brane and outer membrane of nerve being retained.And cireular cells were observed to adhere to and grow steadily between the basal membranes in the ANX+MenSCs group.The result of fluorescence staining suggested MenSCs sur-vived in the scaffold,which secreted nerve growth factor(NGF).At 8 weeks after transplantation,the retraction time of the afected limbs in the ANX+MenSCs group was signifcanly shorter than that in the ANX group(P<0.05),and the climbing slope,SFI and tibialis anterior muscle wet weight ratio were significantly higher than those in the ANX group(P<0.05).There was no signifcant diference in the retraction time of the afected limbs,elimbing gradient and wet weight ratio of the tibialis anterior muscle between the ANX+MenSCs group and the autologous group(P>0.05),but the SFI of the autologous group was significantly higher(P<0.05).Transmission electron microscopy showed that compared with the ANX group,the myelin structure of ANX+MenSCs group and the autologous group was more com-plete,slightly edema,small and

关 键 词：经血源性间充质干细胞 脱细胞异种神经桥接体 共培养体系 坐骨神经缺损修复 大鼠 

分 类 号：R651.3[医药卫生—外科学]