摘要
目的:观察骨髓间充质干细胞(BMSCs)移植大鼠脑内的存活和向血管内皮细胞的分化,以及脑脉通颗粒对其存活和分化的影响。方法:大鼠体外全骨髓贴壁筛选培养和扩增BMSCs;大鼠随机分为假手术组、模型组、移植组、联合组;线栓法制备(MCAO)模型;术后24 h将BMSCs由同侧颈内动脉移植脑内;电镜下观察BMSCs移植后脑组织微血管内皮细胞超微结构变化;检测脑组织中CD31+表达;免疫组化双标法观察Brdu/FⅧ双标细胞变化。结果:与假手术组比较(49.23±6.17),各模型组大鼠脑组织CD31+表达显著减弱(25.14±5.47,15.25±4.50,18.91±3.25)(P<0.01);与模型组比较,移植组14,28 d的CD31+表达增强(30.36±5.74,41.12±6.71)(P<0.01);与移植组比较,各联合组CD31+表达显著增强,以28 d尤为显著(56.42±7.16)(P<0.01)。移植组和联合组大鼠脑内有Brdu标记细胞存活;联合组14,28 d脑内Brdu标记细胞较移植组显著增多(P<0.05,P<0.01)。各移植组与联合组大鼠脑内可见Brdu/Ⅷ双标细胞存活;各联合组大鼠Brdu/Ⅷ双标细胞均较移植组增多(P<0.05,P<0.01)。结论:BMSCs存活的数量随移植脑内时间的延长而呈增加趋势;脑脉通可使移植脑内的BMSCs成活增多。BMSCs在脑内向血管内皮细胞的分化比率随移植后时间的延长呈先增强再减弱的变化趋势;脑脉通可使BMSCs向血管内皮细胞分化和成血管样作用增强。
Objective： To observe the survival and differentiation of bone mesenchymal stem cells （BMSCs） into vascular endothelial cells in brain of cerebral ischemic rats after transplantation and the effect of Naomaitong （NMT） on the survival and differentiation of BMSCs. Method： Rats whole bone marrow were cultivated and BMSCs were purified and increased by methods of adherence and selection in vitro. Rats were randomly divided into different groups. Middle cerebral artery occlusion （MCAO） model was duplicated with nylon thread. Rats of transplantation and combination groups were transplanted with BMSCs via carotid artery at 24 h after operation. Electron microscopic was used to observe the microvascular endothelial cell uhrastructure of brain tissue after BMSCs transplantation. CD31 ＋ expression in brain tissue was detected in this expertment, and the changes of Brdu/FVIII double labeled cells were observed by using the method of immunohistocliemistry double labeling. Result： CD31 ＋ expression was significant in brain of sham operation group, while the expression of CD31 ＋ was significantly decreased in rats of each model group （P 〈 0.01 ）. In brain of 14, 28 d transplantion groups, the expression of CD31 ＋ increased obviously （P 〈 0.01 ）. In comparison with that of transplantation group, CD31 ＋ expression in brain of each combination groups enhanced significantly, especially in 28 d group （P 〈 0.01 ）. In brain of transplantation and combination groups, survival of Brdu mark cells could be detected, and the cells increased obviously in brain of 14, 28 d combination groups compared with that of transplantation group. Survival of the transplanted ceils being labeled doulely by Brdu （Brdu/F VIII ） was observed in transplantation and combination groups, and the cells increased obviously in brain of each combination group than that of transplantation group. Conclusion： It is showed the tendency that the survival of BMSCs in brain tissue increase following the extension of time after transplantation, and NMT could make more BMSCs survive. The ratio of BMSCs differentiating into vascular endothelial cells enhance firstly and then weaken with the trend of the time in rats＇ brain after transplantation. NMT could make the servival of BMSCs enhanced after transplantation and make the differentiation into vascular endothelial ceils angiogenic effect increase.
出处
《中国实验方剂学杂志》
CAS
北大核心
2013年第19期187-192,共6页
Chinese Journal of Experimental Traditional Medical Formulae
基金
国家自然科学基金项目(30973744)
