摘要
目的探讨大气细颗粒物(PM2.5)对原代培养大鼠心肌细胞的急性毒性作用及对缝隙连接通讯的影响。方法对出生24h的SPF级SD大鼠乳鼠分离心肌细胞进行原代培养,以不同浓度(0、1、10、100μg/ml)的PM2.5染毒24h,采用划痕染料标记示踪法(SLTD)测定细胞缝隙连接通讯(GJIC)水平,采用间接免疫荧光细胞化学方法测定缝隙连接蛋白Cx43分布及密度,采用免疫印迹法测定连接蛋白Cx43的表达。结果随PM2.5浓度的上升,细胞间荧光扩散面积减少,细胞膜连接处Cx43绿色荧光减弱,Cx43蛋白表达量稍有减少。与对照组比较,10和100μg/ml染毒组细胞间荧光扩散面积减少,差异有统计学意义(P<0.01);10和100μg/ml染毒组Cx43荧光强度下降,差异有统计学意义(P<0.01)。结论PM2.5可抑制心肌细胞的缝隙连接通讯功能,其机制可能是通过影响心肌细胞的Cx43表达和分布。
Objective To investigate the effects of PM2.5 on gap junctional intercellular communication （GJIC） between rat cardiomyocytes. Methods Primary cultured cardiomyocytes were prepared from 1-day-old Sprague-Dawley rats and exposed to PM2.5（1,10,100 μg/ml）for 24 hours. The GJIC between cardiomyocytes was detected by the scrape loading dye transfer assay. The distribution and density of connexin43（Cx43） in the cells was detected by indirect immunofluorescence and the expression of Cx43 was detected by western blotting. Results The gap junctional intercellular communication between cardiomyocytes was significantly inhibited by PM2.5 in a dose-dependent manner. The fluorescence density of Cx43 was significantly decreased in PM2.5-treated cells,and the expression of Cx43 was also slightly decreased. Conclusion PM2.5 can inhibit GJIC between cardiomyocytes,which may be mediated by the decreased expression and aberrant distribution of Cx43 in PM2.5-treated cells.
出处
《环境与健康杂志》
CAS
CSCD
北大核心
2008年第7期565-568,F0003,共5页
Journal of Environment and Health
基金
国家自然科学基金资助项目(30571534
20637020)
关键词
空气污染
颗粒物
心肌细胞
缝隙连接通讯
缝隙连接蛋白CX43
Air pollution
Particulate matter
Myocardial cell
Gap junctional intercellular communication
Connexin43
