嗅球神经层胶质细胞、嗅黏膜胶质细胞、SC修复周围神经缺损能力比较  

COMPARISON OF COMPETENCE OF OLFACTORY GLOBULAR NERVE LAYER GLIACYTES,OLFACTORY EPITHELIAL GLIACYTES AND SC IN REPAIRING NERVE DEFECT

作　　者：刘夫海[1] 陈统一[1] 张键[1] 陈增淦[1] 

机构地区：[1]复旦大学中山医院骨科,上海200032

出　　处：《中国修复重建外科杂志》2009年第1期1-7,共7页Chinese Journal of Reparative and Reconstructive Surgery

基　　金：国家自然科学基金资助项目(30471744)~~

摘　　要：目的比较嗅黏膜胶质细胞、嗅球神经层(olfactory globular nerve layer,OGNL)胶质细胞及SC修复周围神经缺损的能力,并优选出最适宜移植治疗周围神经缺损的胶质细胞。方法取20只2～3月龄雌性Wistar大鼠,体外培养嗅黏膜胶质细胞、OGNL胶质细胞及SC,调整浓度为1×106个/mL纯化浓缩备用。另将80只成年雌性Wistar鼠,切除坐骨神经25mm轴突,保留神经外膜吻合于近端,制备神经缺损模型。将动物随机分成A、B、C、D4组,每组20只,于神经外膜腔内分别植入10μLDMEM/F12培养液、SC、OGNL胶质细胞以及嗅黏膜胶质细胞。术后3个月,采用踝关节功能评分评估神经缺损的修复效果;行大体、光镜、透射电镜观察神经再生情况;荧光显微镜下观察逆行标记荧光红的运输距离,免疫荧光检测胶质纤维酸性蛋白(glial fibrillary acidic protein,GFAP)及神经生长因子(nerve growth factor,NGF)浓度;ELISA法检测髓鞘碱性蛋白(myelin basic protein,MBP)及神经丝蛋白(neurofilament,NF)浓度。结果踝关节功能评分:A、B、C、D组分别为(3.325±0.963)、(4.200±1.005)、(5.143±0.635)和(5.950±0.154)分,组间比较差异均有统计学意义(P<0.05)。大体、光镜及透射电镜观察示,D组神经缺损再生最完全,C组次之,B组较差,A组最差。荧光红在神经中运行距离,D组最长,C组次之,B组较短,A组最短;NGF及GFAP浓度,D组最高,C组次之,B组较差,A组最低。A、B、C、D组MBP浓度分别为(9.817±3.267)、(12.347±3.091)、(14.937±2.075)和(22.757±0.871)ng/mL,A、B组间差异无统计学意义(P>0.05),其余各组间比较差异均有统计学意义(P<0.05);NF浓度分别为(13.869±5.677)、(18.498±3.889)、(23.443±2.260)、(27.610±1.125)ng/mL,组间比较差异均有统计学意义(P<0.05)。结论嗅黏膜胶质细胞、OGNL胶质细胞、SC均能促进坐骨神经缺损再生,嗅觉系统胶质细胞促进神经再生效果优于SC,而嗅黏膜胶质细胞促进神经再生能力优于OGNL胶Objective To compare their competence of olfactory epithelial gliacytes, olfactory globular nerve layer （OGNL） gliacytes and SC in repair nerve defect of sciatic nerve, and select the best gliacytes for repair of peripheral nerve defect. Methods Olfactory epithelial gliacytes, OGNL gliacytes and SC were extracted from 20 female Wistar rats aged 2-3 months and cultured in vitro for 2 weeks, then purifed and condensed for transplantation. Eighty adult female Wistar rats were randomized into groups A, B, C and D （n=20）. The left sciatic nerves were excised 25 mm axons and retained epineurium lumen anastomosed to proximal ends. The culture mediums, SC, OGNL gliacytes, and olfactory epithelial gliacytes were transplanted into the epineurium lumen of groups A, B, C and D, respectively. Three months postoperatively, the injured sciatic nerve regeneration was evaluated by methods of macroscopic observation, photomicroscope, transmission electron microscope, retro-marked fluorescence transportation distance, the glial fibrillary acidic protein （GFAP） and nerve growth factor （NGF） were assayed by immunofluorescence, and the myelin basic protein （MBP） and neurofilament （NF） protein were assayed by ELISA. Results The scores of ankle joint were （3.325 ± 0.963）, （4.200 ± 1.005）, （5.143 ± 0.635） and （5.950 ± 0.154） in groups A, B, C and D, respectively; showing statistically significant difference between groups （P 〈 0.05）. The observations of gross, sections under microscope and transmission electron microscope showed the regeneration of defect nerve was best in group D, followed by group C, and group B was superior to group A. The transportation distance of retro-marked fluorescence was longest in group D, followed by group C, and group B was superior to group A. The concentrations of GFAP and NGF were largest in group D, followed by group C, and group B was superior to group A. The MBP concentrations were （9.817 ± 3.267）, （12.347 ± 3.091）, （14.937 ± 2.075） and

关 键 词：嗅球神经层胶质细胞 嗅黏膜胶质细胞 SC 细胞移植 坐骨神经损伤 大鼠 

分 类 号：R651.3[医药卫生—外科学]