CAJ | 학술논문

目的研究活性小胶质细胞移植对脊髓损伤后大鼠运动功能恢复的作用.方法选用成年雌性Wistar大鼠40只,随机分为4组,前两组为运动功能观察组,采用自制改良Allen脊髓打击装置打击脊髓,建立中度脊髓损伤的动物模型,分成移植组和对照组,每组10只.后两组为组织学观察组,用相同方法制作动物模型,也分为移植组与对照组,每组10只.在动物模型制作早期进行新生大鼠小胶质细胞的培养、分离以及纯化,并对小胶质细胞进行鉴定和细胞纯度的测定.移植组术后7 d再次暴露损伤部位,微量注射器以损伤部位为中心注射移植活化小胶质细胞悬液.而对照组不移植.运动功能观察组分别在移植后第1天、第1,2,3,4周对大鼠进行运动功能评分;组织学观察组在相同时相点进行Naoumenko及Feigin小胶质细胞石蜡切片染色,每个时间点移植组和对照组每组随机抽取两只大鼠取材切片,镜下观察并计数小胶质细胞.对所得数据进行统计学分析.结果术后1,2,3,4周,对照组与移植组随时间延长BBB评分均逐渐升高,移植组术后2,3,4周后肢运动功能评分较对照组明显升高(P＜0.05);Naoumenko-Feigin染色计数,移植组比同期对照组阳性细胞数显著增多(P＜0.05).结论小胶质细胞移植可促进脊髓损伤大鼠后肢运动功能的恢复.
목적 연구활성소효질세포이식대척수손상후대서운동공능회복적작용.방법 선용성년자성Wistar대서40지,수궤분위4조,전량조위운동공능관찰조,채용자제개량Allen척수타격장치타격척수,건립중도척수손상적동물모형,분성이식조화대조조,매조10지.후량조위조직학관찰조,용상동방법제작동물모형,야분위이식조여대조조,매조10지.재동물모형제작조기진행신생대서소효질세포적배양、분리이급순화,병대소효질세포진행감정화세포순도적측정.이식조술후7 d재차폭로손상부위,미량주사기이손상부위위중심주사이식활화소효질세포현액.이대조조불이식.운동공능관찰조분별재이식후제1천、제1,2,3,4주대대서진행운동공능평분;조직학관찰조재상동시상점진행Naoumenko급Feigin소효질세포석사절편염색,매개시간점이식조화대조조매조수궤추취량지대서취재절편,경하관찰병계수소효질세포.대소득수거진행통계학분석.결과 술후1,2,3,4주,대조조여이식조수시간연장BBB평분균축점승고,이식조술후2,3,4주후지운동공능평분교대조조명현승고(P＜0.05);Naoumenko-Feigin염색계수,이식조비동기대조조양성세포수현저증다(P＜0.05).결론 소효질세포이식가촉진척수손상대서후지운동공능적회복.
Objective To study the role of activated microglia transplantation in recovery of hindlimb locomotor function in rats with spinal cord injury(SCI). Methods A total of 40 female adult Wistar rats were selected and divided into four groups randomly(10 rats in each group).The former two groups were locomotor function observation groups,in which rat model with spinal cord was established by striking with improved self-made Allen's strike equipment to fabricate moderate spinal cord injury and divided into transplantation group and control group.The latter two were histological observation groups,the spinal cord injury model was fabricated by the same above-mentioned method and divided into transplantation and control groups.Before fabricating the spinal cord injury model,the microglia of the newborn rats were cultured,separated,purified and identified and the purity of the microglia determined.The injury position was exposed again seven days after transplantation and the cell suspension of microglia was injected around the injury position with microsyringe,which was free in the control group.The hindlimb locomotor function of rats was detected and scored at 1 day,1,2,3 and 4 weeks in the locomotor function observation groups after transplantation respectively.At the same time,two rats were extracted randomly from the control group and the transplantation group in histological observation groups to cut specimen and slice for Naoumenko-Feign paraffin section and dying.Then,the microglia were observed and counted by microscope and analyzed statistically. Results At 1,2,3 and 4 weeks after operation,the BBB score of the control group and the transplantation group was increased gradually with the time.But compared with the control group,transplantation group had higher scores of hindlimb locomotor function at 2,3 and 4 weeks after operation,with statistical difference(P＜0.05).Naoumenko-Feigin paraffin section and dying and microglia counting showed that positive microglia number in the transplantation group was increased more obviously than the control group,with statistical difference(P＜0.05).Conclusion Activated microglia transplantation can promote the recovery of the hindlimb locomotor function in SCI rats.