摘要
目的观察嗅球成鞘细胞（olfactoryensheathingcells，OECs）在损伤脊髓内的成髓鞘作用及其对髓鞘形成的影响。方法用NYU—II型脊髓撞击器（强度为10g×25mm）损伤大鼠T10脊髓后1周，将绿色荧光蛋白（greenfluorescenceprotein，GFP）-OECs植入脊髓损伤部位及其首尾两侧，设为OECs移植组；以同量改良Eagle培养基（Dulbecco’SmodificationofEagle’Smedi．um，DMEM）注射作为对照组。移植后6周，取出脊髓做冰冻切片，行髓鞘碱性蛋白（myelinbasicprotein，MBP）、0蛋白（proteinzero，PO）和S100蛋白（S100protein，S100）免疫细胞化学染色，定性和半定量观察免疫染色情况；并行塑料包埋后半薄和超薄切片，先于光镜下进行定性和半定量观察，再于电镜下观察髓鞘的超微结构。结果OECs移植组S100、MBP、PO所显示的有髓神经纤维由正常组织向损伤区延伸，并可贯穿损伤区，所显示的纤维分别占损伤区面积的12．3％、11．6％和9．3％，且3种蛋白显示的神经纤维数量差异无统计学意义，但均显著高于对照组（2．89％）（P〈0．01）。OECs移植组半薄切片上移植组损伤区内有髓神经纤维的数量为（354．67±59．00），较对照组的（167．33±42．16）明显增多（P〈0．01），OECs移植组形成的髓鞘较对照组直径小、髓鞘薄。结论OECs可以促进再生神经纤维髓鞘形成，部分OECs自身形成髓鞘，但髓鞘结构较薄。
Objective To detect the myelinating role of olfactory ensheathing cells （OECs） in the contused spinal cord and their impact on remyelinafion. Methods The rats were subjected to spinal cord injury at Ti0 （10 g x 25 mm） using a NYU-II impaetor. One week later, the rats were transplanted with green fluorescence protein （GFP） -OECs （ OECs group） or an equal volume of Dulbeceo＇ s modifi- cation of Eagle＇s medium （DMEM） （control group） at epicenter of the injury as well as its rostral and caudal sites. Six weeks after transplantation, the spinal cords were removed for frozen section. Myelin basic protein （MBP） , protein zero （P0） , and S100 protein （S100） were determined with qualitative and semi-quantitative immunocytochemieal assay. Moreover, plastic embedded semithin and ultrathin sections were prepared for qualitative and semi-quantitative examination under light microscopy and electroscopic study of myelin sheath ultrastructure. Results In OECs group, the nerve fibers labeled with SIO0, MBP, and PO were extended from the normal tissues to the injured region and even grew through the re- gion with space consuming of 12.3% , 11.6% , and 9.3% respectively. Moreover, there were no statisti- cal differences regarding the number of fibers labeled by the three proteins, but all were significantly lar- ger than that in control group （2.89% , P 〈 0. 01 ）. Number of myelinated nerve fibers in injured regions on hemithin sections was increased significantly to 354.67 _＋ 59.00 in OECs group, with significant differ- ence compared with 167.33 ±42. 16 in control group （ P 〈 0. 01 ）. The regenerated myelin sheaths in OECs group were smaller and thicker than those in control group. Conclusions OECs can accelerate regeneration of myelinated nerve fibers. Additionally, some OECs form myelin sheaths themselves, but the sheath structures are relatively thinner.
出处
《中华创伤杂志》
CAS
CSCD
北大核心
2014年第1期57-61,共5页
Chinese Journal of Trauma
基金
国家自然科学基金面上资助项目（81371341）
国家重点基础研究发展计划资助项目（2012CB518106）
创伤、烧伤与复合伤国家重点实验室I类课题资助项目（SKLZZ201003）
关键词
脊髓损伤
髓鞘
嗅球成鞘细胞
Spinal cord injuries
Myelin sheath
Olfactory ensheathing cells
