白藜芦醇对TNF-α诱导的血管内皮细胞炎性反应的影响  

Effect of resveratrol on TNF-α-induced vascular endothelial inflammation in vitro

作　　者：陈明亮[1] 易龙[1] 金鑫[1] 谢琦[1] 周曦[1] 陈春烨[1] 张婷[1] 王丽[1] 糜漫天[1] 

机构地区：[1]第三军医大学军事预防医学院营养与食品安全研究中心,重庆市营养与食品安全重点实验室,重庆市医学营养研究中心,重庆400038

出　　处：《第三军医大学学报》2012年第13期1255-1258,共4页Journal of Third Military Medical University

基　　金：国家自然科学基金面上项目(81172670);重庆市重点实验室资助项目(2006CA1003)~~

摘　　要：目的观察白藜芦醇对TNF-α刺激引起的内皮细胞炎症反应的影响,并围绕TNFR1探索相关作用机制。方法不同浓度(0.01、0.1、1、2.5、5、10、20、50、100、200、400 ng/ml)TNF-α处理人血管内皮细胞株EA.hy926,24 h后MTT法和ELISA法分别检测细胞增殖活力和细胞培养液中sICAM-1释放水平;白藜芦醇(浓度为0.1、1、10μmol/L)预处理24 h后,再用TNF-α(10 ng/ml)处理24 h,利用ELISA法检测细胞培养液中sICAM-1释放水平,qRT-PCR法检测ICAM-1mRNA和TNFR1 mRNA表达水平。结果高浓度TNF-α(≥100 ng/ml)刺激EA.hy926细胞后,细胞增殖活力显著下降(P<0.05),当TNF-浓度为200 ng/ml时,与对照组比较细胞增殖活力下降约59.7%;浓度为10 ng/ml的TNF-α刺激内皮细胞后,细胞的ICAM-1及TNFR1 mRNA表达明显上调(P<0.05);白藜芦醇预处理内皮细胞后可明显下调TNFR1 mRNA表达水平(P<0.05),并显著抑制TNF-α诱导的ICAM-1的表达(P<0.05),且抑制作用随着浓度的增加而增强。结论白藜芦醇可能通过抑制TNFR1表达,进而抑制TNF-α刺激引起的炎性因子释放,减轻内皮炎性损伤。Objective To determine the inhibitory effects of resveratrol on TNF-α induced inflammatory damage in vascular endothelial cell line EA. hy926. Methods After the endothelial EA. hy926 cells were treated with TNF-α of different concentrations （0.01, 0.1,1, 2.5, 5, 10, 20, 50, 100, 200, and 400 ng/ml） for 24 h, cell viability and slCAM-1 release in the supernatant were measured by MTT and ELISA assay, respectively. Cells were pretreated with resveratrol of different concentrations （0.1, 1, and 10 p, mol/L） for 24 h, and then incubated with TNF-a （ 10 ng/ml） for another 24 h. sICAM-1 level in the the supernatant was determined by slCAM-1 ELISA kit. The mRNA expression of intercellular adhesion molecule-1 （ICAM-1） and tumor necrosis factor receptor-1 （TNFR1） were measured by qRT-PCR assay. Results After treated with TNF-α at high concentration （≥ 100 ng/ml）, cell viability was decreased significantly. Cell viability incubated with 200 ng/ml TNF-α was decreased by about 59.7% compared with control. When treated with a low concentration of TNF-α （ 10 ng/ml ） , the mRNA levels of ICAM-1 and TNFR1 were increased markedly （P 〈0.05 ）. However, resveratrol pretreatment significantly （P 〈 0. 01 ） inhibited the increase of ICAM-1 and TNFR1 mRNA expression induced by TNF-α in a dose-dependent manner. Conclusion Resveratrol might inhibit the TNF-α-induced inflammatory cytokine release and alleviate endothelial inflammation by inhibiting TNFRl-mediated signaling pathway. atherosclerosis ; resveratrol ; EA. hy926 ; inflammation ; intercellular adhesion molecule-1 ; tumor necrosis factor receptor-1

关 键 词：动脉粥样硬化 白藜芦醇 血管内皮细胞 炎性反应 ICAM-1 TNFR1 

分 类 号：R151.2[医药卫生—营养与食品卫生学] R364.5[医药卫生—公共卫生与预防医学]