| AMPK激动剂AICAR通过阻滞细胞周期于G0/G1期抑制肺动脉平滑肌细胞增殖 |
| 投稿时间：2014-09-17 修订日期：2014-10-16 点此下载全文 |
| 引用本文：官文俊,许臣洪.AMPK激动剂AICAR通过阻滞细胞周期于G0/G1期抑制肺动脉平滑肌细胞增殖[J].医学研究杂志,2015,44(5):111-113 |
| DOI：
10.11969/j.issn.1673-548X.2015.05.030 |
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| 中文摘要:目的 建立PDGF-BB诱导的原代大鼠肺动脉平滑肌细胞(pulmonary vascular smooth muscle cells,PASMCs)的增殖细胞模型,通过AMPK激动剂AICAR干预,探讨AICAR对PASMCs细胞周期和增殖的影响及其机制,为肺血管重构防治寻找靶点。方法 通过20ng/ml PDGF-BB刺激诱导PASMCs增殖建立细胞模型,采用AICAR(0.5mmol/L)干预PDGF-BB诱导的PASMCs增殖,Western blot法检测总的和磷酸化的AMPK, CCK-8检测PASMCs增殖,流式细胞仪分析细胞周期,实时定量PCR(RT-PCR)检测cyclinD1、cyclinE、CDK2/4/6 mRNA的表达。结果 Western blot法检测表明AICAR可以活化AMPK,CCK-8检测结果表明AICAR能够抑制PDGF-BB诱导的PASMCs增殖;流式细胞仪检测结果表明AICAR能够抑制细胞周期于G0/G1期,RT-PCR结果表明AICAR可以抑制cyclinD1、cyclinE、CDK2、CDK4和CDK6的mRNA的表达。结论 AICAR通过抑制cyclinD1、cyclinE、CDK2/4/6 的mRNA表达阻滞细胞周期于G0/G1~S期,抑制PASMCs增殖。 |
| 中文关键词:AMPK PDGF-BB 肺动脉平滑肌细胞 细胞周期 增殖 |
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| AICAR Blocks the Proliferation of Pulmonary Artery Smooth Muscle Cells by Inhibition of Cell Cycle in G0/G1 |
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| Abstract:Objective To establish the PDGF-BB-induced proliferation of rat pulmonary artery smooth muscle cells(PASMCs), and investigate the effects of the AMPK agonist AICAR on the cell cycle of PASMCs, in order to search new drugs for prevention of pulmonary vascular remodeling. Methods 20ng/ml PDGF-BB was used to induced the proliferation of PASMCs, and the effect of 0.5mmol/L AICAR on the proliferation of PASMCs was observed. Western blot was used to detect the total and phosphorylated AMPK. The proliferation of PASMCs was determined by CCK-8. The mRNA expression of cyclinD1, cyclinE and CDK2/4/6 were detected by flow cytometry analysis cell cycle,quantitative real-time PCR. Results Western blot results indicated AICAR could promote the activation of AMPK. CCK-8 test results showed that AICAR blocked the proliferation of PASMCs induced by PDGF-BB. Flow cytometry analysis indicated that AICAR arrested the cell cycle in G0/G1 to S phase. RT- PCR results demonstrated that AICAR inhibited the mRNA expression of cyclinD1, cyclinE and CDK2/4/6. Conclusion The AMPK agonist AICAR can block the proliferation of PASMCs induced through arrest cell cycle in G0/G1-S phase by regulation the mRNA expression of cyclin D1, cyclinE, CDK2/4/6, and AICAR has a potential application in preventing pulmonary vascular remodeling. |
| keywords:AMPK PDGF-BB Pulmonary artery smooth muscle cells Cell cycle Proliferation |
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