杨桂棠 韩雅玲△ 闫承慧 田孝祥 李洋.过表达CREG抑制ox-LDL诱导的人血管平滑肌细胞增殖[J].,2015,15(36):7028-7032 |
过表达CREG抑制ox-LDL诱导的人血管平滑肌细胞增殖 |
Over-expression of CREG Inhibits ox-LDL Induced Proliferation of HumanVascular Smooth Muscle Cells |
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DOI: |
中文关键词: CREG 血管平滑肌细胞 增殖 动脉粥样硬化 |
英文关键词: CREG Vascular smooth muscle cell Proliferation Atherosclerosis |
基金项目:辽宁省博士启动基金项目(20131136) |
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中文摘要: |
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60
滋g/mL 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BrdU
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
BrdU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
BrdU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BrdU 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。 |
英文摘要: |
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 滋g/mL ox-LDL. At different time points (0 h, 12 h, 24 h and 48 h), cell numbers were counted
to plot growth curve; BrdU 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, BrdU 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 BrdU 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 BrdU positive cells, increased CREG expression while down-regulated phosphorylated Erk1/2.Conclusion:Over-expression of CREG could inhibit ox-LDL induced proliferation of human VSMC possibly through down-regulating Erk1/2
phosphorylation. |
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