【摘要】 目的研究贴壁培养的神经前体细胞(NPCs)间隙连接蛋白43(Cx43)的表达及酒精对其表达的影响。方法采用贴壁培养的方法,取孕14 d的胎鼠端脑进行NPCs体外培养,经形态学观察和纯度鉴定后,再采用免疫荧光化学方法,观察NPCs中Cx43的表达,利用体外NPCs酒精损伤实验模型,蛋白质免疫印迹技术和刻痕标记/荧光扩散实验,分析酒精对NPCs中Cx43表达及间隙连接功能的影响。结果原代贴壁培养7 d的细胞NPCs阳性率达86.74%,免疫荧光化学法证实NPCs能表达Cx43,蛋白质免疫印迹实验证实酒精可以减少Cx43的表达,刻痕标记/荧光扩散实验显示酒精对NPCs的间隙连接功能具有抑制作用。结论原代贴壁培养的NPCs可大量表达Cx43,酒精可以通过降低Cx43表达量抑制间隙连接功能,并且该抑制作用随酒精作用浓度的增加而提高。更多还原

【Abstract】 Objective To investigate expression of connexin 43(Cx43) in neural precursor cells(NPCs) and to explore the effect of ethanol on its expression.Methods Primary adherent cultures of NPCs isolated from the brain of embryonic rats at pregnancy 14d were used in this study.After observing morphology of cells and identifying purity of the NPCs,we investigated expression of Cx43 by an immunofluorescence technique.Then,we studied the influence of ethanol on expression of Cx43 and on function of gap junctions by Western-blot and Scrape-loading dye in cell injury models induced by ethanol.Results The positive rate of NPCs in primary cultures that had been cultured by an adherent culture method for 7 days was 86.74%.Immunofluorescence technique showed that NPCs expressed Cx43.Western-blot showed that expression level of Cx43 gradually decreased with an increase of ethanol concentration.Scrape-loading dye revealed that ethanol inhibited the function of gap junctions.Conclusion NPCs cultured by an adherent culture method strongly express Cx43.Ethanol inhibits the function of gap junctions by decreasing expression level of Cx43.The inhibitory effect is increased with an increase of ethanol concentration.更多还原