摘要
目的:探讨神经营养因子-3(NT-3)联合神经干细胞(NSC)移植促进缺血缺氧性脑损伤乳鼠的神经功能恢复情况。方法:进行大鼠胎脑皮层组织NSC的体外培养并制成单细胞悬液,将60只出生7 d的Wistar大鼠采用随机数字表法将其分为正常组(A组)、缺血缺氧模型组(B组)、单纯NSC移植组(C组)、NT-3联合NSC移植组(D组)各15只。B、C、D组采用经典模型方法制备缺血缺氧模型,3 d后行平衡木试验,A组老鼠不给予任何干预措施。B组制模型后第三日移植生理盐水,C组行NSC移植,D组行NT-3和NSC共同移植。移植后4周取鼠脑行大体标本、HE染色、免疫组织化学检查,比较各组结果。结果:①体外培养新生鼠脑皮层组织可见团状细胞球,Nestin染色提示阳性,说明是NSC。②大体标本比较,C组及D组则较B组血供明显增多、脑组织萎缩、塌陷程度明显减轻。③HE染色,B组侧脑室周围细胞肿胀、变性、坏死,排列紊乱,局部囊性变和炎性细胞聚积;C组组织结构破坏以及细胞形态较脑瘫组有所改善,囊性病变减少;D组恢复情况更为明显。④免疫组织化学,B组较正常组胶质细胞阳性数增加,少突胶质细胞阳性细胞数减少;C组较B组胶质细胞有所减少,少突胶质细胞较B组增多。D组恢复更好。结论:NT-3能够通过促进NSC的增值和诱导分化作用更进一步地促进缺血缺氧脑损伤患者的神经功能恢复。
Objective： To determine the effect of neurotrophin-3 combined with neural stem cell graft in neu- rological function recovery of hypoxic-ischemic brain injury in suckling rats. Methods： Rat fetal cerebral cortical tissue NSCs was cultured in vitro and passaged, then cultured neural stem cells were suspended. 60 suckling rats were be divided into A, B, C and D, four groups by radom number： normal group （A） , hypoxia-ischemia model group （B）, the simple neural stem cell transplantation group （C）, NT-3 joint neural stem cell transplantation group （D）. The classic ischemia and hypoxia model has been used for the balace beam test 3 days later： group A weren＇t given any intervention, group B were given saline on the third day after surgery, group C were transplanted by NSCs and the mice of group D were transplanted by NSC together with NT-3. 4 weeks after transplantation, the brains were removed for gross specimens, HE, immunohistochemical staining pathological comparison. Results ： ① The bulk muhicelluar spheroids in cortex graft in vitro cultured rats＇ tiss were neural stemcell according to positive Nestin staining. ②Compared to group B, the blood supply of grop C and D increased, and the degree of the brain tiss atrophying and collapsing decreased. ③Besause of HE staining, the cells around lateral ventricle in group Bwere swelling, degeneration, necrosis, arranged disorder, local cystic changes and inflammatory cells accumula- tion. Compared to structural damage, the damage degree of normal tissue in group C a improved, which was better in group D. ④In immune past, the neuroglia positive cells in group B were more than that in group A, and oligo- dendroglia cells in group B were less. Compared to group B, the neuroglia cells in group C were less and oligoden- droglia cells in group C were more. The situation of group D was the best. Conclusion： NT-3 can promote the re- covery of neurological function of hypoxic-ischemic brain injury patients by promoting neural stem cell proliferation and differentiation
出处
《新医学》
2013年第3期206-210,共5页
Journal of New Medicine
基金
国家自然科学基金(30960393
81160155)
