大鼠脊髓损伤后对巢蛋白及突触素I表达的影响  

Effect on the expression of nestin and synapsin I after spinal cord injury in rats

作　　者：申福国 孔维丽[2] 刘士臣[1] 崔绪民 崔宏宇[1] 赵中南[1] 张富运[1] 宋春雨[1] 杨建华[1] 

机构地区：[1]佳木斯大学第一附属医院骨科,黑龙江省佳木斯154002 [2]佳木斯大学第一附属医院内分泌科,154002 [3]鹤北林业局医院外科,154002

出　　处：《中华急诊医学杂志》2014年第12期1348-1354,共7页Chinese Journal of Emergency Medicine

基　　金：黑龙江省自然基金（D201160）;黑龙江省教育基金（12511539）

摘　　要：目的探讨同种异体骨髓单个核细胞（BMMNCs）移植对脊髓全横断损伤区局部微环境中巢蛋白和突触素I表达的影响及意义。方法120只清洁级Winstar大鼠（雌雄不拘）随机（随机数字法）取87只分为3组。假手术组29只：切除相应的椎板不损伤脊髓；模型组29只：建立脊髓全横断模型；细胞移植组29只：建立脊髓全横断模型后，给予BMMNCs移植治疗。各组大鼠分别于5d、1周、2周、4周利用免疫组织化学技术、RT-PCR检测脊髓组织中巢蛋白（nestin）与突触素I（SynapsinⅠ）的表达变化及利用MIAS-2000图像处理系统分析观察nestin和SynapsinⅠ阳性产物在脊髓的分布，并测量阳性产物的吸光度值。采用SPSS17．0统计分析数据，组间比较，方差齐者采用完全随机设计资料的方差分析，样本问的两两比较用q检验；方差不齐者用成组设计多个样本比较的秩和检验（Kruskal-Wallis法），样本问两两比较的秩和检验（Nemenyi法）；P〈0．05为差异具有统计学意义。结果免疫组织染色、RT-PCR及图像处理系统分析结果显示模型对照组在伤后约5d可见损伤区域附近nestin表达升高，2周达高峰，4周后nestin表达明显下调，细胞移植组在脊髓伤5d可见损伤区域附近nestin表达升高，1周达高峰，4周后nestin表达明显下调，且各时期与假手术组、模型组相比较差异均具有统计学意义（P〈0．05）。损伤5d后，在损伤区域出现突触素I的表达，但模型组、细胞移植组突触素I的表达量在各时间点均低于假手术组，细胞移植组高于模型组（P〈0．01）。结论同种异体的骨髓单个核细胞移植可能改变了局部的微环境并上调了脊髓内巢蛋白和突触素I的表达。Objective To investigate the effects and significance of site transplantation of allogenic bone marrow mononuclear cells （BMMNCs） on the expression of nestin and synapsin I in the local mieroenvironment of transected injury zone in spinal cord. Methods From a pool of 120 clean grade Wistar rats （ Irrespective of gender） , 87 rats were randomly （ random number） selected and subdivided into three groups. Sham operation group （n = 29 ） had lamineetomy done without inducing spinal cord injury. The model control group （n = 29 ） had laminectomy done followed by induction of complete spinal cord transeeted injury. Cell transplantation group （n = 29） had BMMNCs transplantation initiated after induction of complete spinal cord transected injury. Nestin and Synapsin expressions in all rats of each group were assessed on day 5, week 1, week 2 and week 4, respectively by using immunohistochemical and RT-PCRtechniques. Spread of Nestin positive and Synapsin I positive products along the spinal cord were assessed by use of MIAS-2000 image processing system analysis. Optical density of these products was then measured. Statistical data analysis was done using SPSS17. 0. Uniform variance analysis was done for inter group comparison using completely random design data, while paired sample comparison done with the Q test. For non-uniform variance, multiple samples were compared using design rank sum test with Kruskal-Wallis method, whereas paired samples rank sum test with Nemenyi method. P value of P 〈 0. 05 was regarded statistically significant, and P 〈 0.01 regarded highly statistically significant. Results Immunohistochemistry, RT-PCR and image processing system analysis showed model control group had increased nestin expression within the damaged zone 5 days after injury, reaching peak level at 2 weeks then significantly down regulated 4 weeks later. In cell transplantation group, nestin expression around spinal cord injury zone increased 5 days after injury and reached the peak level at 1 week, t

关 键 词：骨髓单个核细胞 脊髓全横断损伤 细胞移植 巢蛋白 突触素I 脊髓损伤 免疫组化 反转录酶-聚合酶链锁反应 

分 类 号：R651.2[医药卫生—外科学]