CAJ | 학술논문

目的观察基质细胞衍生因子-1(SDF-1)对神经干细胞(NSCs)迁移的影响.方法由GFP转基因SD大鼠胚胎脑组织获取NSCs并进行传代培养,免疫细胞化学染色法检测SDF-1特异性受体CXCR4的表达,利用Blind-Well小室体外迁移体系观察不同浓度的SDF-1(0、1、10、50、100、500、1000μg/L)对NSCs定向迁移数量的影响,随后分别使用CXCR4激动剂和阻断剂处理NSCs,再次利用上述方法观察最适浓度SDF-1时NSCs的迁移.结果成功分离培养得到能够稳定表达GFP的NSCs,且CXCR4在该种NSCs上有表达.体外趋化实验结果表明,SDF-1对NSCs有较强的趋化作用,随着SDF-1浓度的升高,发生迁移的细胞数量也随之增加,并于SDF-1浓度为500μg/L时达到最高峰;CXCR4特异性激动剂和阻断剂分别能够增强和减弱SDF-1对NSCs定向迁移的趋化作用.结论 SDF-1与其特异性受体CXCR4相互作用,能够对NSCs的定向迁移产生靶向性作用.
목적 관찰기질세포연생인자-1(SDF-1)대신경간세포(NSCs)천이적영향.방법 유GFP전기인SD대서배태뇌조직획취NSCs병진행전대배양,면역세포화학염색법검측SDF-1특이성수체CXCR4적표체,이용Blind-Well소실체외천이체계관찰불동농도적SDF-1(0、1、10、50、100、500、1000μg/L)대NSCs정향천이수량적영향,수후분별사용CXCR4격동제화조단제처리NSCs,재차이용상술방법관찰최괄농도SDF-1시NSCs적천이.결과 성공분리배양득도능구은정표체GFP적NSCs,차CXCR4재해충NSCs상유표체.체외추화실험결과표명,SDF-1대NSCs유교강적추화작용,수착SDF-1농도적승고,발생천이적세포수량야수지증가,병우SDF-1농도위500μg/L시체도최고봉;CXCR4특이성격동제화조단제분별능구증강화감약SDF-1대NSCs정향천이적추화작용.결론 SDF-1여기특이성수체CXCR4상호작용,능구대NSCs적정향천이산생파향성작용.
Objective To investigate the effect of stromal cell derived factor-1 ( SDF-1) on the regulation of neural stem cells (NSCs) migration. Methods NSCs were obtained from the cerebral cortex of embryonic GFP transgenic rats. After cell cultured in vitro, CXCR4, specific receptor of SDF-1, was detected by fluorescence immunocytochemistry. Using Blind-Well chambers, the chemotactic effects of SDF-1 was investigated by counting the cells which had crossed 8μm pore membrane and adhered to the superficies inferia of the membrane when confronted with varying concentrations of SDF-1α (0, 1, 10, 50, 100, 500 and 1000 μg/L) and the agonist or antagonist of CXCR4. Results Neurospheres were formed, which expressed GFP and were capable of differentiating into neurons (β-tubulin + ) and astrocytes (GFAP + ) in media without mitogens. Fluorescence immunocytochemistry showed that CXCR4 and Nestin were co-expressed in NSCs. SDF-1 showed great chemotaxis to NSCs, and the amount of cells migrating through the membrane in 500 μg/L SDF-1 group was more than that in other groups (P < 0. 05). The number of cells crossing the membrane could be augmented and diminished by the agonist and antagonist of CXCR4 respectively. Conclusion SDF-1 binding to its specific receptor CXCR4 was capable of inducing NSCs migrating directionally to the source of SDF-1.