摘要
目的:探讨E1A激活基因阻遏子(Cellular repressor of E1A-stimulated genes,CREG)在氧化型低密度脂蛋白(Oxidized low density lipoprotein,ox-LDL)引起的人血管平滑肌细胞(Vascular smooth muscle cell,VSMC)增殖中的作用及可能机制。方法:用60μg/m L ox-LDL处理人胸主动脉VSMC,在不同时间点(0 h,12 h,24 h,48 h)计数细胞并绘制生长曲线;在48 h时间点进行Brd U染色检测增殖细胞的比率;采用Western Blot检测各时间点CREG表达。通过向VSMC中转染携带CREG基因的质粒载体,筛选获得过表达CREG的VSMC(VSMCCREG)。进一步用ox-LDL处理VSMCCREG,采用前述方法绘制生长曲线、进行48 h时间点Brd U染色并检测CREG和磷酸化Erk1/2的表达。结果:与未经处理的对照组VSMC相比,ox-LDL处理的VMSC细胞总数及Brd U阳性细胞比率显著增加;并且,CREG表达随处理时间延长逐渐降低。此外,与ox-LDL处理的VSMC组相比,ox-LDL处理的VSMCCREG组细胞总数及Brd U阳性细胞比率均显著下降;并且CREG表达增加、磷酸化Erk1/2水平下降。结论:过表达CREG可能通过降低Erk1/2磷酸化抑制ox-LDL诱导的人VSMC增殖。
Objective： To investigate the role and possible mechanism of cellular repressor of E1A-stimulated genes（CREG） in oxidized low density lipoprotein（ox-LDL） induced proliferation of human vascular smooth muscle cells（VSMC）. Methods： VSMC from human thoracic aorta were treated with 60 μg/m L ox-LDL. At different time points（0 h, 12 h, 24 h and 48 h）, cell numbers were counted to plot growth curve; Brd U staining was performed to assess percentage of proliferative cells at 48 h; Western Blot was carried out to detect CREG expression at different time points. Then VSMC over-expressing CREG（VSMCCREG） was established by transfecting VSMC with a plasmid vector carrying CREG gene. These cells were also treated with ox-LDL. Cell counting, Brd U staining, and detection of CREG and phospho-Erk1/2 were performed as mentioned above. Results： Compared to untreated VSMC, VSMC treated with ox-LDL had more total cells and higher ratio of Brd U positive cells; moreover, expression of CREG decreased gradually after treatment. In addition, compared to VSMC treated with ox-LDL, VSMCCREG treated with ox-LDL showed significantly less number of total cells,decreased ratio of Brd U positive cells, increased CREG expression while down-regulated phosphorylated Erk1/2. Conclusions：Over-expression of CREG could inhibit ox-LDL induced proliferation of human VSMC possibly through down-regulating Erk1/2phosphorylation.
出处
《现代生物医学进展》
CAS
2015年第36期7028-7032,共5页
Progress in Modern Biomedicine
基金
辽宁省博士启动基金项目(20131136)
