微囊化兔雪旺细胞移植后脊髓半横断损伤大鼠血清MBP动态变化及组织学观察

发布时间：2018-03-26 14:42

本文选题：微囊　切入点：雪旺细胞　出处：《南昌大学》2006年硕士论文

【摘要】：目的观察大鼠脊髓半横断损伤后植入微囊化兔雪旺细胞(Schwann Cells，SCs)对血清髓鞘碱性蛋白(Myelin Basic Protein，MBP)变化的影响及组织学变化，初步探讨微囊化异种SCs移植治疗脊髓损伤(Spinal Cord injury，SCI)的修复机制及对髓鞘再形成的影响。方法取健康成年家兔10只，体重2200±200g；Sprague-Dawley(SD)大鼠142只，体重220±20g，雌雄不拘，随机分为4组：A组(单纯损伤组，n=44)；B组(SCs移植组，n=44)；C组(微囊化SCs移植组，n=44)；D组(正常对照组，n=10)。无菌条件下取成年家兔双侧坐骨神经干，利用反复差速贴附法体外培养SCs，制备SCs细胞悬液及微囊化SCs细胞悬液。建立大鼠T_(10)脊髓右半横断损伤模型，A、B、C组分别在损伤处移入吸附10μl的DMEM培养液、SCs悬液、微囊化SCs悬液的明胶海绵。D组为正常组。动物在灌注前，，心脏穿刺采血，用ELISA法检测血清MBP。术后1d、3d、7d、14d、28d，各组每时相取8只(D组取2只)大鼠灌注固定，取损伤周围处脊髓组织做石蜡切片，行HE、Loyez氏法髓鞘染色。另外，A、B、C组2w、8w时相点各2只大鼠，取损伤周围处脊髓组织固定，Epon816包埋，醋酸铀和柠檬酸铝双染，超薄切片电镜观察。结果各组损伤后1d即出现血清MBP升高，7d达到高峰，以后逐渐恢复。移植后1d、3d各组间血清MBP水平差异无显著性(P＞0.05)。移植后7d、14d、28d，与A、B组比较，C组血清MBP含量均较低(P＜0.05)，尤14d明显(P＜0.01)；A组与B组比较，除术后7d血清MBP含量比较差异有显著性(P＜0.05)外，其他时相的比较差异无显著性(P＞0.05)。SCI后1d、3d各组损伤侧脊髓神经元变性坏死，数量减少，部分神经元胞体皱缩、核固缩，细胞边界模糊，髓鞘数量减少、结构松驰，脊髓组织内片状出血，出血周围可见神经胶质细胞和巨噬细胞浸润，但B组和C组较少见。7d时坏死区有胶质细胞增生和白细胞浸
[Abstract]:Objective to observe the effect of microencapsulated Schwann cells of Schwann cells (Schwann Cellsberg SCS) on the serum myelin Basic protein (MBP) and its histological changes in rats after spinal cord hemitransection injury. To explore the repair mechanism of microencapsulated xenogeneic SCs transplantation for spinal cord injury (Spinal Cord injuryant) and its effect on myelin reformation. Methods Ten healthy adult rabbits, weighing 2200 卤200g Sprague-Dawley SD rats, weighing 220 卤20g, were selected. They were randomly divided into 4 groups: group A (simple injury group), group B (SCs transplantation group), group C (microencapsulated SCs transplantation group), group D (normal control group). Bilateral sciatic nerve trunk of adult rabbits was removed under aseptic condition. SCs cell suspensions and microencapsulated SCs cell suspensions were prepared by repeated differential attachment method in vitro. A rat model of right half transection of spinal cord was established. The microencapsulated SCs suspension of gelfoam group D was used as normal group. Blood samples were collected from heart puncture before perfusion, and serum MBP was detected by ELISA method. The rats were perfused and fixed with 8 rats in each group at 1 day, 3 days, 7 days, 14 days and 28 days after operation, and 8 rats in each group were taken from each group. The spinal cord tissue around the injury was taken as paraffin section and stained with Hefer Loyez's myelin sheath. In addition, two rats in group A and B (group C) were treated with Epon816 entrapped with fixed Epon816 at 8 weeks after injury. The specimens were stained with uranium acetate and aluminum citrate, and the ultrathin sections were observed by electron microscope. Results the serum MBP level increased 1 day after injury and reached the peak at 7 days after injury. There was no significant difference in serum MBP levels between the three groups on the 3rd day after transplantation (P > 0.05). The serum MBP levels in group C were lower than those in group A (P < 0.05), and those in group A were significantly lower than those in group B (P < 0.01) on the 7th day after transplantation (P < 0.01), and those in group B were significantly lower than those in group B (P < 0.01). Except for the significant difference in serum MBP content on the 7th day after operation (P < 0.05), there was no significant difference in other phases (P > P > 1 d after 0.05).SCI). The number of neurons in the injured side of the spinal cord was decreased, the number of neurons was reduced, some of the neurons were corrugated, nuclear pyknosis, and the cell boundaries were blurred. The number of myelin decreased, the structure relaxed, the lamellar hemorrhage of spinal cord tissue, the glial cells and macrophages infiltrated around the hemorrhage, but in group B and C there were glial cell proliferation and leucocyte leucocytosis in the necrotic area on the 7th day.
【学位授予单位】：南昌大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R651.2;R361

【参考文献】