神经营养因子-3联合神经干细胞移植对乳鼠缺血缺氧性脑损伤神经修复作用

Neurotrophin-3 plus neural stem cells transplantation for nerve repair in rats with hypoxic-ischemic brain injury

目的观察神经营养因子-3(neurotrophin-3,NT-3)联合神经干细胞(neural stem cells,NSCs)移植对乳鼠缺血缺氧性脑损伤后的神经修复作用。方法 NSCs传代培养,7日龄Wistar乳鼠60只,随机分为对照组10只与实验组50只,实验组制备缺血缺氧性脑损伤模型后存活37只,随机分为脑损伤组12只、NSCs组12只与NT-3联合NSCs组(联合组)13只。模型制备成功3d后,联合组左侧侧脑室移植NSCs与NT-3,NSCs组移植NSCs,脑损伤组移植PBS,对照组空针插入后不做移植。4组于移植3、7周时取脑组织行组织病理学检查,采用免疫印迹法观察髓鞘碱性蛋白表达水平。结果移植3、7周时脑损伤组损伤侧脑室旁正常组织结构破坏,出现多个大小不一的囊腔;NSCs组损伤侧脑室旁正常组织结构有所恢复,囊变少见;联合组损伤侧脑室周围组织结构较其他组恢复好;移植3、7周时,联合组、NSCs组侧脑室周围脑组织髓鞘碱性蛋白表达量高于脑损伤组与对照组(P<0.05),脑损伤组高于对照组(P<0.05);联合组移植3周时侧脑室周围脑组织髓鞘碱性蛋白表达量高于NSCs组(P<0.05),移植7周时与NSCs组比较差异无统计学意义(P>0.05)。结论 NT-3可促进NSCs分化为少突胶质细胞,有助于NSCs移植治疗乳鼠缺血缺氧性脑损伤的效果。

Objective To observe the effect of the transplantation of neurotrophin 3 （NT-3） plus neural stem cells （NSCs） on nerve repair in neonatal rats with hypoxic-ischemic brain injury （HIBD）. Methods NSCs were subculture＆ Sixty 7-day old Wistar rats were randomly divided into experimental group （n=50） and control group （n= 10）. Thirty seven survived rats in experimental group after the establishment of HIBD animal models were randomly divided into brain injury group （n=12）, NSCs group （n=12） and NT-3＋NSCs group （n=13）. Three days after the models were established, the left lateral ventricle was transplanted NSCs and NT 3 in NT-3 ＋ NSCs group, NSCs in NSCs group, and PBS in brain injury group. Control group was inserted empty needle without transplantation. The brain tissue was taken out for pathological examination in 3 and 7 weeks after transplantation in 4 groups. Western blot was used to observe the expression of myelin basic protein. Results In 3 and 7 weeks after transplantation, adjacent normal tissue structure of left lateral ventricle was damaged, appearing cavity with different sizes in brain injury group, was restored with rare cystic degeneration in NSCs group, and recovered better in NT-3 ＋ NSCs group. The expression of myelin basic protein was higher in NT-3＋NSCs group and NSCs group than that in brain injury group and control group （P〈0.05）, higher in brain injury group than that in control group （P^0.05）. The expression of myelin basic protein was higher in NT-3＋ NSCs group than that in NSCs group in 3 weeks after transplantation （P〈0.05）, and there was no significant difference in 7 weeks after transplantation （P 〉 0.05 ）. Conclusions NT-3 can promote the differentiation of NSCs into oligodendrocytes, which improves the therapeutic effect of NSCs transplantation on ischemic hypoxic brain in rats.