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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-2mRNA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 (SFN) on the expression of cyclooxygenase-2 and iNOS mRNA in vascular endothelial cells (ECV304) induced by lipopolysaccharide (LPS) and its transcriptional mechanism. Methods The ECV304 cells were cultured in vitro. The expression of COX-2 mRNA was determined by real-time PCR. The expression of iNOS mRNA was detected by RT-PCR. The expression of IκB -α protein expression; nuclear translocation of NF-κB was measured by immunofluorescence hybridization. Results Real-time PCR results showed that 5 ~ 20μmol / L SFN could significantly reduce the expression of COX-2 mRNA induced by lipopolysaccharide, the inhibition rates were 24%, 16% and 30% respectively. RT- Induced iNOS mRNA expression was also significantly inhibited; SFN inhibited LPS induced IκB-α protein degradation and NF-κB nuclear translocation. Conclusion SFN can inhibit LPS-induced vascular endothelial cell inflammatory response, suggesting that SFN has a preventive effect on cardiovascular disease.