摘要
目的观察NgR在新生大鼠少突胶质前体细胞系(OLPs)的表达和细胞定位,以及缺氧缺血(OGD)损伤后的表达变化,探讨其在OLPs损伤后再生抑制中的作用。方法采用改良振荡分离纯化法体外培养OLPs,采用其特异性抗体A2B5、O4和O1作细胞鉴定;用NgR抗体检测其在OLPs表达。用含连二亚硫酸钠的无糖培养基制作OLPs OGD模型,MTT法检测各组细胞存活率;用免疫荧光法和免疫印迹法观察NgR在细胞OGD后的变化。结果OLPs在分离纯化培养后的第1,2和4天均表达NgR,阳性表达位于胞体及突起;细胞OGD后10min,NgR表达出现增强,30min后明显升高,与对照组比较差异具有显著性(均P(0.05);MTT结果显示,OGD30min,细胞存活率较对照组显著降低(P(0.05)。结论OLPs胞体及突起上均表达NgR;OGD后NgR表达明显增强,细胞存活率显著降低,提示NgR可能参与OGD后OLPs的再生抑制过程。
Objective This study examined the NgR expression in oligodendrocyte precursor cells （OLPs） and its changes after oxygen ＆ glucose deprivation （OGD） in order to explore the role of NgR expression in the regeneration of OLPs after OGD in neonatal rats. Methods The OLPs from 2-day-old neonatal rats were separated by improved separation and purification through agitation and then cultured in chemically defined medium. OLPs OGD model was prepared using the medium consisting of Na2S2O4 and Earle＇s fluid in vitro. Immunofluorescence assay was applied to identify the OLPs with its specific antibodies such as A2B5, 04 and O1. Western blot was used to detect the NgR expression in OLPs 10 and 30 minutes after OGD. The livability rate of cells was detected by MTT. Results NgR expression was found in both the cell body and the prominence of purified OLPs. NgR expression in OLPs increased significantly 10 and 30 minutes after OGD compared with that in OLPs without OGD （ controls, P 〈 0.05 ）. MTT showed that the livability rate of OLPs at 30 minutes following OGD was significantly lower than that of controls （65.97 ± 3.69% vs 97.17± 6.88%, P 〈 0.05 ）. Conclusions NgR is expressed in both the cell body and the prominence of OLPs. NgR expression increases while cell livability decreases following OGD, suggesting that NgR may play a role in the inhibition of regeneration of OLPs.
出处
《中国当代儿科杂志》
CAS
CSCD
2007年第5期445-448,共4页
Chinese Journal of Contemporary Pediatrics
基金
教育部博士点基金项目资助(编号:20050610071)
