摘要
目的探讨香烟烟雾对血管壁组织中的两种主要细胞:血管平滑肌细胞和血管内皮细胞增殖活力的影响。方法应用磷酸盐缓冲液(PBS)作为溶剂萃取香烟烟雾,制备烟雾萃取剂。用此烟雾萃取剂依据不同浓度对以上两种细胞进行染毒处理,CCK-8试剂盒测定细胞增殖活力,Western blot检测转录因子c-jun、细胞周期蛋白D1(cyclinD1)的变化,转染过表达c-jun质粒检测cyclinD1的变化。结果 CCK-8检测显示在烟雾染毒浓度(0.625%~10%)范围内,平滑肌细胞增殖活力增加,内皮细胞在染毒处理后随浓度增加其活力降低。染毒处理48 h后,平滑肌细胞和内皮细胞的染毒处理组的c-jun表达与PBS对照组相比差异均无统计学意义,平滑肌细胞染毒组的p-c-jun比PBS对照组明显增加(P<0.05),内皮细胞染毒组的p-c-jun降低(P<0.05)。细胞周期蛋白cyclinD1在平滑肌细胞染毒处理后明显增加(P<0.05),内皮细胞染毒后则显著降低(P<0.05),并且过表达c-jun后cyclinD1的表达水平也相应增高。结论烟雾染毒具有增加血管平滑肌细胞增殖活力和降低内皮细胞活力的作用,其与c-jun磷酸化和cyclinD1表达有关。
Objective To investigate the cell viabilities of vascular smooth muscle cells and vascular endothelial cells stimulated by cigarette smoke extract (CSE). Methods The CSE was prepared by smoke-bub- bled phosphate buffered saline (PBS) generation. After culturing cells with different concentrations of CSE, we used the cell counting kit-8 to determine the cell viability. The expression levels of c-jun and cyclinD1 were ana- lyzed through Western blot. The c-jun plasmid was transfected to detect the change of cyclinD1 expression. Results The smooth muscle cell viability increased when the CSE concentration ranged O. 625% -10%, whereas the endothelial cells viability decreased when exposed to the CSE concentration. After exposure to CSE for 48 hours, there was no difference in c-jun expression between toxin group and PBS group; however, the expression of p-c-jun in the smooth muscle cells significantly increased in the toxin groups than in the PBS group (P 〈 0. 05 ) and the expression of p-c-jun in the vascular endothelial cells significantly decreased (P 〈 0. 05 ). The level of cyclinD1 significantly increased after exposed to CSE, and its expression level also increased in respond to thec-jun overexpression. Conclusion CSE can enhance the proliferation of vascular smooth muscle cells and de- crease in the activity of endothelial cells proliferation, which may be explained by the phosphorylation of c-jun and the expression of cyclinD1.
出处
《中国医学科学院学报》
CAS
CSCD
北大核心
2013年第2期185-189,共5页
Acta Academiae Medicinae Sinicae