论文部分内容阅读
目的探讨胰岛素样生长因子-1(IGF-1)基因修饰神经干细胞(NSCs-IGF-1)移植对缺氧缺血性脑损伤(HIBD)新生大鼠脑功能恢复的治疗作用。方法采用结扎大鼠左侧颈总动脉,低氧箱中缺氧处理的方法,制成新生大鼠HIBD模型,随机分为正常对照组、模型对照组、神经干细胞(NSCs)移植组和NSCs-IGF-1移植组,尾静脉注入法进行干细胞移植。分别于移植1 d、7 d、14 d、21 d取5只大鼠脑组织进行病理切片,通过5-溴脱氧尿嘧啶(BrdU)、巢蛋白(Nestin)、神经元特异性烯醇化酶(NSE)免疫组织化学染色观察NSCs在宿主脑内存活及增殖和分化情况,并通过Y-迷宫实验和运动功能检测对剩余大鼠脑功能恢复情况进行观察。结果移植7 d后,NSCs移植组和NSCs-IGF-1移植组均可见到BrdU阳性细胞,二组比较差异有统计学意义(P<0.05)。在观测时间点内,NSCs-IGF-1移植组Nestin阳性细胞表达量早期升高,移植后14 d达峰值,后逐渐下降;NSE阳性细胞表达量逐渐升高。模型对照组在移植7 d后Nestin及NSE阳性细胞表达量逐渐降低,2组比较差异均有统计学意义(Pa<0.01)。移植14 d,NSCs-IGF-1移植组Nestin及NSE阳性细胞表达量均明显高于NSCs移植组(Pa<0.01)。大鼠学习记忆功能检测及运动功能检测结果均显示NSCs-IGF-1移植组明显优于模型对照组及NSCs移植组,二者差异均具有统计学意义(Pa<0.01,0.05)。结论 NSCs及NSCs-IGF-1可以在脑内存活、增殖及分化,并可部分促进缺氧缺血大鼠脑功能的恢复;NSCs-IGF-1较单独移植NSCs效果好。
Objective To investigate the therapeutic effect of insulin-like growth factor-1 (IGF-1) gene-modified neural stem cells (NSCs-IGF-1) transplantation on brain function recovery in neonatal rats with hypoxic-ischemic brain damage (HIBD). Methods HIBD models of neonatal rats were made by ligating left common carotid arteries in rats and hypoxia in hypoxia chambers. They were randomly divided into normal control group, model control group, neural stem cell (NSCs) transplantation group and NSCs- IGF-1 transplantation group, tail vein injection for stem cell transplantation. Five rat brain tissues were harvested at day 1, day 7, day 14, and day 21, respectively, for histopathology. The brain tissues were stained with 5-bromodeoxyuridine (BrdU), Nestin and neuron specific enolase NSE) were used to observe the survival, proliferation and differentiation of NSCs in the host brain. The recovery of brain function of the remaining rats was observed by Y-maze test and motor function test. Results BrdU positive cells were observed in NSCs transplantation group and NSCs-IGF-1 transplantation group after 7 days of transplantation. The difference between the two groups was statistically significant (P <0.05). At the observation time point, the expression of Nestin positive cells in NSCs-IGF-1 transplantation group increased at early stage and peaked on the 14th day after transplantation, and then decreased gradually. The expression of NSE positive cells increased gradually. The expression of Nestin and NSE positive cells in model control group decreased gradually after 7 days of transplantation, the differences between the two groups were statistically significant (Pa <0.01). At 14 days after transplantation, the expression of Nestin and NSE positive cells in NSCs-IGF-1 group were significantly higher than that in NSCs transplantation group (Pa0.01). The learning and memory function tests and motor function test results showed that NSCs-IGF-1 transplantation group was significantly better than the model control group and NSCs transplantation group, the difference was statistically significant (Pa <0.01,0.05). Conclusion NSCs and NSCs-IGF-1 can survive, proliferate and differentiate in the brain and partially promote the recovery of brain function in hypoxic-ischemic rats. NSCs-IGF-1 is more effective than NSCs alone.