番茄红素上调细胞周期素依赖性激酶4和细胞周期素D1的表达促进人外周血内皮祖细胞周期进展

Effects of Lycopene on Cell Cycle and Expression of Cyclin-dependent Kinase 4 and Cyclin D1 in Endothelial Progenitor Cells of Human Peripheral Blood

摘要:

目的探讨番茄红素对人外周血内皮祖细胞活性、周期调控的作用及其分子机制。方法体外原代培养人外周血内皮祖细胞,采用噻唑蓝法、流式细胞仪、逆转录聚合酶链反应、实时聚合酶链反应和Western blot方法检测番茄红素对内皮祖细胞活性、细胞周期分布的影响及细胞周期相关蛋白细胞周期素依赖性激酶4和细胞周期素D1的表达情况。结果噻唑蓝比色实验显示,不同浓度(0.01、0.1、1μmol/L)番茄红素能显著增加内皮祖细胞的生长能力,且呈一定的量效与时效关系;流式细胞仪分析显示,番茄红素呈浓度依赖性使内皮祖细胞的G0/G1期细胞比例减少,S期细胞比例增多;逆转录聚合酶链反应、实时聚合酶链反应和Western blot分析表明,番茄红素呈浓度依赖性使内皮祖细胞中细胞周期素依赖性激酶4和细胞周期素D1表达上调。结论番茄红素对内皮祖细胞的促进生长作用,可能与上调细胞周期素依赖性激酶4、细胞周期素D1的表达,使G0/G1期细胞比例减少有关。

Abstract:

Aim To investigate the effect of lycopene in endothelial progenitor cells of human peripheral blood and its underlying molecular mechanism. Motheds The promotion survival effect of lycopene in different concentrations on endothelial progenitor cells was measured by thiazolyl blue(MTT) assay.Cell cycle was analyzed by flow cytometry.The expression levels of cyclin-dependent kinase 4(CDK4) and cyclin D1 were detected by reverse transcription polymerase chain reaction(RT-PCR) and real-time polymerase chain reaction(real-time PCR) and Western blot. Results The MTT assay showed that lycopen promoted survival effect on endothelial progenitor cells in a time and dose-dependent manner.Flow cytometry analysis revealed that G1 phase of endothelial progenitor cells was decreased after exposed to different concentrations of lycopene.The expression levels of CDK4 and cyclin D1 were increased significantly after treated with lycopene in a dose-dependent manner using RT-PCR,real-time PCR and Western blot. ConclusionThe promotion survival effect of lycopene in endothelial progenitor cells is related to G1 phrase reducing through increased expression levels of CDK4 and cyclin D1.