论文部分内容阅读
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 effect of E1A-stimulated genes repressor (CRA) on the expression of Vascular smooth muscle cell (VSMC) induced by Oxidized low density lipoprotein (ox-LDL) The role of proliferation and possible mechanism. Methods: Human VSMCs were treated with 60 μg / mL ox-LDL and the growth curves were counted at different time points (0 h, 12 h, 24 h, 48 h). BrdU staining was performed at 48 h The ratio of proliferating cells was detected. Western blot was used to detect the expression of CREG at each time point. The VSMC overexpressing CREG (VSMCCREG) was screened by transfecting the VSMC with the plasmid vector carrying the CREG gene. The growth curve of VSMCCREG was further treated with ox-LDL, and the growth curve was drawn by the above method. The Brd U staining at 48 h and the expression of CREG and phosphorylated Erkl / 2 were detected. Results: Compared with untreated VSMC, ox-LDL-treated VMSC cells and BrdU positive cells increased significantly. And CREG expression decreased gradually with the prolongation of treatment time. In addition, compared with the VSMC group treated with ox-LDL, the total number of cells and BrdU positive cells in VSMCCREG treated with ox-LDL decreased significantly, and the expression of CREG increased and the phosphorylated Erk1 / 2 decreased. Conclusion: Overexpression of CREG may inhibit ox-LDL-induced proliferation of human VSMC by decreasing Erk1 / 2 phosphorylation.