| 建立以HIF-1α为靶标的高通量筛选防治动脉粥样硬化先导化合物的细胞模型 |
| 投稿时间：2018-06-11 修订日期：2018-09-25 点此下载全文 |
| 引用本文：钱俞君,秦春霞,孙莉莉,丁华敏,李铁军.建立以HIF-1α为靶标的高通量筛选防治动脉粥样硬化先导化合物的细胞模型[J].药学实践杂志,2019,37(1):27~31 |
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| 基金项目:上海市浦东新区科技发展基金（PKJ2015-Y26）；上海市浦东新区卫生和计划生育委员会学科建设（PWZbr2017-16） |
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| 中文摘要:目的 建立一个高通量筛选防治动脉粥样硬化先导化合物的体外细胞模型。方法 克隆HIF-1α低氧反应元件（hypoxia response element，HRE）至荧光素酶报告基因表达载体pGL3-Enhancer，构建荧光素酶表达载体pGL3-HIF-1α-HRE，转染人单核细胞THP-1并筛选稳定表达细胞株THP-1-HIF-1α-HRE。结果 Real Time-PCR检测表明低氧培养可以有效上调THP-1-HIF-1α-HRE细胞HIF-1α蛋白表达和荧光素酶活性，而洛伐他汀和姜黄素预处理可以有效抑制低氧引起的THP-1-HIF-1α-HRE细胞内HIF-1α蛋白表达及荧光素酶活性。结论 成功建立了筛选抗动脉粥样硬化的先导化合物体外高通量细胞模型THP1-HIF-1α-HRE。 |
| 中文关键词:缺氧诱导因子1α 高通量 荧光素酶活性 动脉粥样硬化 筛选 |
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| A cell model for high-throughput screening lead compounds targeting HIF-1α for atherosclerosis treatment |
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| Abstract:Objective To establish a high-throughput in-vitro screening cell model for anti-atherosclerosis leading compounds. Methods Hypoxia response element (HRE) was cloned into a luciferase reporter vector, pGL3-Enhancer, to construct pGL3-HIF-1α-HRE. The THP-1 human monocyte cell line was infected with the pGL3-HIF-1α-HRE and a stable cell line, THP-1-HIF-1α-HRE, was screened. Results Real-time PCR assay showed that HIF-1α expression and luciferase activity in THP-1-HIF-1α-HRE cells was effectively upregulated by hypoxia. The increase of HIF-1α expression and luciferase activity induced by hypoxia was significantly inhibited by lovastatin or curcumin. Conclusion THP1-HIF-1α-HRE, an in-vitro cell model for high-throughput screening lead compounds for anti-atherosclerosis (AS) was successfully established. |
| keywords:hypoxia inducible factor-1α high-throughput luciferase atherosclerosis screen |
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