Yang Ching Jen, Lin Chia Yi, Hsieh Tze-Chen, Olson Susan C, Wu Joseph M
Department of Biochemistry & Molecular Biology, New York Medical College Valhalla, New York 10595, USA.
Am J Cardiovasc Dis. 2011;1(1):16-30. Epub 2011 Apr 26.
Population studies suggest that moderate red wine intake correlates with reduced risk of cardiovascular disease (CVD); cardioprotection may attribute to consumption of red wine polyphenol resveratrol. Since inflammation plays a key role in CVD, we investigated modulation of inflammation by resveratrol and its metabolites by determining the expression and release of chemokine, eotaxin-1, in cultured human pulmonary artery endothelial cells (HPAEC) treated with proinflammatory cytokines IL-13 and TNF-α. Up-regulation of eotaxin-1 gene expression by IL-13 or TNF-α was confirmed by RT-PCR, by reporter assays using eotaxin-1 gene promoter constructs, and by the changes in transcriptional factors STAT6 and NF-κB. Exposure to resveratrol suppressed IL-13 and TNF-α induced eotaxin-1 gene expression as well as attenuated the eotaxin-1 promoter activity, in coordination with inhibition of expression of JAK-1, reduction in phosphorylated-STAT6 and decreased p65 subunit of NF-κB. In addition, quantitative determination of eotaxin-1 release using enzyme-linked immunosorbent assay (ELISA) showed increased eotaxin-1 release in response to treatment by IL-13 and TNF-α, which was effectively inhibited by resveratrol. Whether resveratrol metabolites affected eotaxin-1 was also tested; piceatannol showed potency similar to resveratrol. We propose that control of eotaxin-1 expression and release by proinflammatory cytokines in HPAEC may be considered as an in vitro model for screening and discovering polyphenols with anti-inflammatory activities and cardioprotective potentials.
人群研究表明,适量饮用红酒与心血管疾病(CVD)风险降低相关;心脏保护作用可能归因于红酒中的多酚白藜芦醇。由于炎症在心血管疾病中起关键作用,我们通过测定促炎细胞因子IL-13和TNF-α处理的培养人肺动脉内皮细胞(HPAEC)中趋化因子嗜酸性粒细胞趋化因子-1(eotaxin-1)的表达和释放,研究了白藜芦醇及其代谢产物对炎症的调节作用。通过RT-PCR、使用eotaxin-1基因启动子构建体的报告基因测定以及转录因子STAT6和NF-κB的变化,证实了IL-13或TNF-α对eotaxin-1基因表达的上调作用。白藜芦醇处理可抑制IL-13和TNF-α诱导的eotaxin-1基因表达,并减弱eotaxin-1启动子活性,同时抑制JAK-1表达、降低磷酸化STAT6水平并减少NF-κB的p65亚基。此外,使用酶联免疫吸附测定(ELISA)对eotaxin-1释放进行定量测定显示,IL-13和TNF-α处理后eotaxin-1释放增加,而白藜芦醇可有效抑制这种增加。我们还测试了白藜芦醇代谢产物是否影响eotaxin-1;紫铆因显示出与白藜芦醇相似的效力。我们提出,HPAEC中促炎细胞因子对eotaxin-1表达和释放的控制可被视为一种体外模型,用于筛选和发现具有抗炎活性和心脏保护潜力的多酚。