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
摘要点击次数: 350
全文下载次数: 242
作者单位E-mail
钱俞君 上海市浦东新区浦南医院药剂科, 上海 200125  
秦春霞 上海市浦东新区浦南医院药剂科, 上海 200125  
孙莉莉 上海市浦东新区浦南医院药剂科, 上海 200125  
丁华敏 上海市浦东新区浦南医院药剂科, 上海 200125  
李铁军 上海市浦东新区浦南医院药剂科, 上海 200125 18930502906@163.com 
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)
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
 
A cell model for high-throughput screening lead compounds targeting HIF-1α for atherosclerosis treatment
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
查看全文  查看/发表评论  下载PDF阅读器
关闭

分享按钮