摘要
为探讨镉对大鼠肝细胞(BRL 3A)缝隙连接通讯功能的影响,以0、2.5、10μmol/L醋酸镉溶液分别处理BRL 3A细胞12h,采用实时分析技术测定细胞指数,显微镜观察细胞形态,划痕染料标记示踪法检测细胞缝隙连接通讯(GJIC),激光共聚焦观察缝隙连接蛋白Cx43的分布,免疫印迹测定Cx43的表达及其磷酸化水平。结果显示,随着镉浓度的增大(2.5、10μmol/L),细胞指数值降低,细胞间荧光黄两侧扩散距离极显著减少(P<0.01),Cx43在细胞膜上分布减少、胞内增多,Cx43的表达及其磷酸化水平极显著降低(P<0.01)。结果表明,镉对BRL 3A细胞有明显细胞毒性,可抑制GJIC,其机制可能是镉诱导BRL 3A细胞Cx43分布异常、表达减少及磷酸化水平降低。
To investigate the effects of cadmium on gap junctional intercellular communication(GJIC)of BRL 3Acells,BRL 3Acells were treated with cadmium(0,2.5and 10μmol/L,respectively)for 12 h.RTCA was used to determine the cell index.The GJIC between BRL 3Acells was detected by the scrape loading dye transfer assay.The distribution of Cx43 was observed using confocal laser scanning microscopy.The Cx43 levels and Cx43 phosphorylation were measured by Western-blotting.In result,as the concentration of cadmium increased,the GJIC was inhibited by cadmium(P<0.01)between adjacent cells.Cx43 immunofluorescent staining was distributed mostly in the intracellular region rather than the membrane.Cx43 levels and Cx43 phosphorylation decreased(P<0.01).In conclusion,cadmium has toxic effects to BRL 3Acells.Cadmium inhibits GJIC between cells,which may be mediated by aberrant distribution of Cx43,decreasing the expression of Cx43 and the phosphorylation of Cx43.
出处
《中国兽医科学》
CAS
CSCD
北大核心
2014年第10期1073-1078,共6页
Chinese Veterinary Science
基金
国家自然科学基金资助项目(31302058
31372495)
