异丙酚干预下人羊膜间充质干细胞移植对大鼠脑损伤的影响
Propofol intervention on mesenchymal stem cell transplantation on brain injury in rats between Human amniotic
张建军 1王东1
作者信息
- 1. 天津市 第四中心医院 神经外科,天津 300000
- 折叠
摘要
目的 人羊膜间充质干细胞(AM-MSCs)移植同时予以异丙酚治疗,观察两者对脑损伤大鼠恢复的影响.方法 健康SD鼠80只,雌雄各半,体质量300～350 g.体外复苏并培养人羊膜间充质干细胞(CM-Dil标记)移植前备用.采用液压颅脑损伤仪,给予液压冲击力,制成重型液压颅脑损伤模型,伤后6 h给予相应治疗随机分成4组:损伤组(培养液移植组)、AM-MSCs移植组、异丙酚组、AM-MSCs+异丙酚组.利用CM-Dil标记在荧光显微镜下观察各组的阳性细胞数.移植后4周各组随机处死6只,并用RT-PCR、Western Blot检测脑组织中GAP-43、AQP4基因表达和蛋白合成的变化.移植后24 h,3天及1、2、3、4周行动物神经学缺损评分,在移植后4周进行Morris水迷宫(Morris Water Maze,MWM)试验.四周后处死行免疫组化、HE染色.结果 CM-Dil的阳性细胞数AM-MSCs+异丙酚组最多,异丙酚组、AM-MSCs移植组次之,损伤组最少,且各组之间差异有显著性意义(P<0.05).移植后4周脑损伤周围组织AQP4及其mRNA的表达损伤组高于AM-MSCs移植组、异丙酚组;AM-MSCs移植组、异丙酚组高于AM-MSCs+异丙酚组(P<0.05);GAP-43及其mRNA的表达损伤组低于AM-MSCs移植组、异丙酚组,AM-MSCs移植组、异丙酚组低于AM-MSCs+异丙酚组(P<0.05).移植后1周,大鼠神经功能障碍评分AM-MSCs+异丙酚组低于异丙酚组、AM-MSCs移植组;异丙酚组、AM-MSCs移植组低于对照组(P<0.05).AM-MSCs+异丙酚组穿越平台次数高于异丙酚组、AM-MSCs移植组(P<0.05),明显高于损伤组(P<0.01).观察伤后4周病理切片,损伤组未见神经轴索通过,异丙酚组、AM-MSCs移植组可见少量神经轴索样结构,AM-MSCs+异丙酚组可见较多神经轴索样结构.结论 异丙酚联合人羊膜间充质干细胞移植治疗大鼠脑损伤可明显改大鼠的神经学功能.
Abstract
Objective Human amniotic mesenchymal stem cells (AM-MSCs) transplantation and to investigate the effect of propofol treatment, recovery of brain injury in rats. Methods 80 SD rats, male and female, weight 300-350 g. Human amniotic mesenchymal stem cells cultured in vitro and recovery (CM-Dil) before transplantation standby. The fluid percussion brain injury device, give 2.5-3.0 ATM hydraulic impact, severe fluid percussion brain injury model was made. After injury 6h give corresponding treatment which were randomly divided into 4 groups: injury group (medium injection group), AM-MSCs transplantation group and propofol group, AM-MSCs + propofol group. The number of positive cells in each group was observed under the fluorescence microscope using CM-Dil markers. 4 weeks after transplantation, 6 rats were randomly executed, and RT-PCR and Blot Western were used to detect the changes of GAP-43, AQP4 gene expression and protein synthesis in brain tissues. After transplantation, 24 h, 3 days and 1, 2, 3, 4 weeks for animal neurological deficit score, in 4 weeks after transplantation of Morris water maze (Morris Water Maze, MWM) test. They were killed after four weeks by immunohistochemistry, HE staining. Results Number of CM-DIL positive cell was the most in AM-MSCs + propofol group, which was in propofol group and AM-MSCs transplantation group were lower than that in the AM-MSCs + propofol group, which in injury group was the least, and the differences among the groups had statistically significant (P<0.05). 4 weeks after transplantation, AQP4 and its mRNA expression of brain damage surrounding tissue in injury group were higher than those in AM-MSCs transplantation group and propofol group; which in AM-MSCs transplantation group and propofol group was higher than those in AM-MSCs + propofol group (P< 0.05); GAP-43 protein and mRNA expression in injury group were lower than those in AM-MSCs transplantation group and propofol group, which in AM-MSCs transplantation group and propofol group were lower than those in AM-MSCs + propofol group (P<0.05). 1 weeks after transplantation, rat nerve dysfunction score in AM-MSCs + propofol group was lower than that in propofol group and AM-MSCs transplantation group; which in propofol group and AM-MSCs transplantation group was lower than that in the control group (P<0.05). The times of crossing platform in AM-MSCs + propofol group was higher than that in propofol group and AM-MSCs transplantation group, significantly higher than that in the injury group (P<0.01). The pathological section was observed 4 weeks after injury, there was no axonal passage in the injury group. In propofol group and AM-MSCs transplantation group, a few axonal like structures were observed, more axonal like structures were observed in AM-MSCs + propofol group. Conclusion Propofol combined with human amniotic mesenchymal stem cells transplantation can significantly improve the neurological function of rats with brain injury.
关键词
人羊膜间充质干细胞/移植/异丙酚/大鼠/脑损伤/功能Key words
human amniotic mesenchymal stem cells/transplantation/propofol/rat/brain injury/function引用本文复制引用
出版年
2017
NETL
NSTL
维普
万方数据