CAJ | 학술논문

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 (Synapsin Ⅰ)的表达变化及利用MIAS-2000图像处理系统分析观察nestin和Synapsin Ⅰ阳性产物在脊髓的分布,并测量阳性产物的吸光度值.采用SPSS 17.0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,样本间的两两比较用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),样本间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.
목적 탐토동충이체골수단개핵세포(BMMNCs)이식대척수전횡단손상구국부미배경중소단백화돌촉소Ⅰ표체적영향급의의.방법 120지청길급Winstar대서(자웅불구)수궤(수궤수자법)취87지분위3조.가수술조29지:절제상응적추판불손상척수;모형조29지:건립척수전횡단모형;세포이식조29지:건립척수전횡단모형후,급여BMMNCs이식치료.각조대서분별우5d、1주、2주、4주이용면역조직화학기술、RT-PCR검측척수조직중소단백(nestin)여돌촉소Ⅰ (Synapsin Ⅰ)적표체변화급이용MIAS-2000도상처리계통분석관찰nestin화Synapsin Ⅰ양성산물재척수적분포,병측량양성산물적흡광도치.채용SPSS 17.0통계분석수거,조간비교,방차제자채용완전수궤설계자료적방차분석,양본간적량량비교용q검험;방차불제자용성조설계다개양본비교적질화검험(Kruskal-Wallis법),양본간량량비교적질화검험(Nemenyi법);P＜0.05위차이구유통계학의의.결과 면역조직염색、RT-PCR급도상처리계통분석결과현시모형대조조재상후약5d가견손상구역부근nestin표체승고,2주체고봉,4주후nestin표체명현하조,세포이식조재척수상5d가견손상구역부근nestin표체승고,1주체고봉,4주후nestin표체명현하조,차각시기여가수술조、모형조상비교차이균구유통계학의의(P＜0.05).손상5d후,재손상구역출현돌촉소Ⅰ적표체,단모형조、세포이식조돌촉소Ⅰ적표체량재각시간점균저우가수술조,세포이식조고우모형조(P＜0.01).결론동충이체적골수단개핵세포이식가능개변료국부적미배경병상조료척수내소단백화돌촉소Ⅰ적표체.
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 microenvironment 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 laminectomy done without inducing spinal cord injury.The model control group (n =29) had laminectomy done followed by induction of complete spinal cord transected 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 nestinexpression 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,then subsequently significantly down regulated 4 weeks later.At each interval,there were significantly statistical differences between sham surgery group and model control group and cell transplantation group (P ＜ 0.05).Synapsin Ⅰ expression appeared within the damaged area 5 days after injury,but at each interval,the expression of synapsin Ⅰ in the model control group and cell transplantation group was lower than that in sham operation group.However,synapsin Ⅰ in cell transplantation group was higher than that in model control group (P ＜ 0.01).Conclusions Allogenic bone marrow mononuclear cells transplantation may alter the local microenvironment and up-regulate the expression of nestin and synapsin Ⅰ in the spinal cord injury model.