莱菔硫烷对脂多糖诱导血管内皮细胞中COX-2和iNOS mRNA表达的影响及转录机制研究  

Effects of sulforaphane on the expression of COX-2 and iNOS mRNA induced by LPS in vascular endothelial cells

作　　者：赵瑞芳[1] 单毓娟[1] 林娜[1] 袁超[1] 王舒然[1] 

机构地区：[1]哈尔滨医科大学公共卫生学院营养与食品卫生教研室,黑龙江哈尔滨150081

出　　处：《毒理学杂志》2009年第1期36-39,共4页Journal of Toxicology

基　　金：国家自然科学基金项目(30571558);黑龙江省自然科学基金(D2005-44)

摘　　要：目的研究莱菔硫烷（SFN）对脂多糖（LPS）诱导的血管内皮细胞ECV304中COX-2和iNOS mRNA表达的影响及转录机制。方法以体外培养的血管内皮细胞ECV304为研究模型；采用实时定量PCR（real-time PCR）测定COX-2 mRNA的表达；采用RT-PCR测定细胞中iNOS mRNA的表达；采用Western blotting测定细胞浆中IκB-α蛋白表达；采用免疫荧光杂交法测定NF-κB核内转位情况。结果Real—time PCR结果表明，5～20μmol／L SFN能明显降低脂多糖诱导的COX-2 mRNA的表达，抑制率分别为24％、16％和30％；RT-PCR结果显示，SFN对LPS诱导的iNOS mRNA表达也有显著的抑制作用；SFN抑制LPS诱导的IκB-α蛋白降解及NF—κB核内转位。结论SFN能抑制LPS诱导的血管内皮细胞炎症反应，提示SFN对心血管疾病具有预防作用。Objective To investigate the effect of sulforaphane on the expression of COX-2 and iNOS mRNA induced by LPS in vascular endothelial cells. Methods Human vascular endothelial ECV 304 cells were cultured in vitro. COX-2 mRNA was detected by real-time reverse transcription polymerase chain reaction （PCR）;iNOS mRNA was measured by RT-PCR; The expression of IκB-α protein in cytoplasm was determined by Western blotting analysis; Nuclear translocation of NF-κB was tested by hybridization. Results 5, 10, 20 μmol/L sulforaphane decreased the expression of COX-2 mRNA by 24%, 16% and 30% by realtime PCR method. Also sulforaphane significantly suppressed the expression of iNOS from the result of RT-PCR. Sulforaphane upregulated the expression of IκB-α protein, an inhibitor of NF-κB, reduced by LPS. Sulforaphane inhibited the nuclear translocation of NF-κB stimulated by LPS in ECV304 cells. Conclusion SFN inhibited the inflammation of ECV304 cells induced by LPS, suggesting its preventive role in cardiovascular diseases.

关 键 词：莱菔硫烷 COX-2 INOS 血管内皮细胞 

分 类 号：Q78[生物学—分子生物学] R541[医药卫生—心血管疾病]