神经营养因子3联合神经干细胞移植治疗缺氧缺血性脑损伤的实验研究

Neural stem cells combined with NT-3 enhancing brain repair in hypoxic-ischemic brain injury

目的观察神经营养因子3(NT-3)与神经干细胞(NSCs)联合移植对缺氧缺血性脑损伤(HIBD)大鼠的治疗效果。方法 Wistar大鼠按随机数字表法分为正常组、单纯脑瘫组、神经干细胞移植组、NT-3联合NSCs移植组。取新生24 h的Wistnr大鼠脑皮质NSCs进行培养、鉴定。选出生7 d的Wistar大鼠制备HIBD动物模型。3 d后行损伤侧侧脑室移植。移植3周后进行神经电生理检测,后取脑组织进行HE染色。结果新生大鼠脑皮层可成功培养出NSCs,并表达NSCs的标志物神经巢蛋白(Nestin),NSCs可分化为神经元及星形胶质细胞。神经电生理:复合肌肉动作电位波幅脑瘫组3.844±1.217 mV,移植组5.218±1.976 mV,NT-3联合NSC组7.512±2.152 mV,差异有统计学意义(P<0.01)。潜伏期:脑瘫组5.729±0.755 ms,移植组5.197±0.709 ms,NT-3联合NSC组4.638±0.720 ms,差异有统计学意义(P<0.01)。HE:脑瘫组脑室旁正常组织结构破坏,可见多个不规则囊腔,神经干细胞移植组脑室旁正常组织结构有所恢复,囊变少见,NT-3移植组组织结构恢复更好。结论 NT-3联合神经干细胞移植是治疗脑瘫的有效办法。

Objective This work was undertaken to determine the effect of neumtrophin - 3 combined with neural stem cell graft in treat- ment of cerebral palsy in new born rats. Methods Neural stem cells （NSC） were obtained from gray tissue of brain of rats born within 24 h, and they were cultured and then identified. A total of 30 Wistar rats which were subjected for hypoxic - ischemic brain injury （HIBI） were randomly assigned to NSC grafts group, NT -3 combined with NSC group and cerebral palsy （CP） group. Three days after the induction of HI, rats in NT - 3 group were grafted with neural stem cells and NT - 3, rats in NSC group received only NSC, and rats in CP group received PBS. Motor and somatosensory evoked potential in lower limb tests were performed at fourth week after transplantation. Hematoxylin and eosin staining was per- formed later. Results Action potential of hind limb quadriceps compound muscle in rats of CP groups showed lower wave amplitude compared with that of graft group （ P 〈 0.05 ）. Hematoxylin and eosin staining showed irregular cells around the ventricle, as well as periventricular leukomalacia in control group. However the tissue recovery was better in rats of graft group. Conclusion NT - 3 combined with NSC transplantation is an effec- tive way for curing cerebral palsy.