姜黄素对脂多糖诱导人脐静脉内皮细胞炎症反应的影响及分子机制

Effect of curcumin on inflammatory response induced by lipopolysaccharide in human umbilical vein endothelial cells and its molecular mechanisms

目的研究姜黄素对脂多糖(LPS)诱导人脐静脉内皮细胞(HUVEC)炎症因子表达的影响,初步探讨可能的分子机制。方法使用LPS(1μg)和姜黄素(5μmol/L、15μmol/L或30μmol/L)处理HUVEC细胞24 h,收集细胞总RNA,实时定量PCR(RT-PCR)和ELISA方法检测细胞TNF-α、MCP-1水平;Western blot检测细胞TLR4、MAPKs、NF-κB p65、磷酸化-NF-κB p65及NF-κB抑制蛋白IκBα的表达情况。结果与空白对照组比较,姜黄素能显著抑制LPS诱导HUVEC细胞TNF-α、MCP-1和TLR4的过表达(P<0.05),明显降低NF-κB p65和MAPKs的磷酸化及IκBα蛋白降解(P<0.05)。结论姜黄素可能基于抑制TLR4/NF-κB,从而影响LPS诱导HUVEC细胞炎症因子的表达。

Objective To investigate the effect of curcumin on the expression of inflammatory response induced by lipopolysaccharide（LPS） in human umbilical vein endothelial cells（HUVEC） and to explore its molecular mechanisms. Methods The to tal RNA was extracted from HUVEC treated with LPS（1 μg/L） and curcumin（5, 15, or 30 μmol/L） for 24 h. The levels of tumor necrosis factor-α（TNF-α） and monocyte chemoattractant protein-1（MCP-1）were detected using real-time reverse transcriptase polymerase chain reaction（RT-PCR） and enzyme-linked immunosorbent assay（ELISA）. Western blot was used to detect the activation of Toll like receptor 4（TLR4）, mitogen-activated protein kinases（MAPKs）, nuclear factor kappa B（NF-κ B） p65, phosphor-NF-κ B p65 and nuclear factor of kappa light polypeptide gene enhancer in B-cells inhibitor alpha（IκBα） in the cells. Results Compared with the untreated cells, curcumin significantly inhibited the overexpression of TNF-α, MCP-1 and TLR4 in HUVEC cells induced by LPS（P0.05）, which also significantly reduced NF-κB p65 and MAPKs phosphorylation and IκBα protein degradation. Conclusion Curcumin suppresses LPS-induced inflammatory response in HUVEC in vitro via inhibiting the TLR4/NF-κB pathways.