盐酸苄丝肼对脂多糖所致人脐静脉内皮细胞炎症的影响及其机制研究

Effects and molecular mechanism of benserazide hydrochloride on LPSinduced inflammation in human umbilical vein endothelial cells

通过脂多糖(LPS)诱导人脐静脉内皮细胞(HUVECs)建立体外炎症模型,进一步验证盐酸苄丝肼的抗炎作用,并探究盐酸苄丝肼抗炎抗动脉粥样硬化的相关分子机制。实验分为空白组(PBS+0.5%FBS DMEM培养基)、模型组[LPS(500μg/m L)+0.5%FBS DMEM培养基]及给药组[LPS(500μg/m L)+盐酸苄丝肼+0.5%FBS DMEM培养基],采用Western blot、ELISA和q PCR检测HUVECs细胞中炎症因子血清淀粉样蛋白P(SAP)、TNF-α、MCP-1蛋白和mRNA的表达水平。采用Western blot检测p65/p-p65、p38/p-p38、IκBα/p-IκBα和AKT/p-AKT的表达水平及p65、p38、IκBα的入核情况。结果显示,盐酸苄丝肼(1×10^(-9)、1×10^(-10)、1×10^(-11)mol/L)能显著抑制促炎细胞因子SAP、TNF-α和MCP-1的蛋白及其mRNA的表达,在下调p65/p-p65、p38/p-p38、IκBα/p-IκBα和AKT/p-AKT的蛋白表达的同时抑制p65、p38、IκBα的核转位,从而抑制相关基因的转录活性。

In this article,human umbilical vein endothelial cells(HUVECs)were induced by lipopolysaccharides(LPS)to establish an in vitro inflammation model to further verify the anti-inflammation effects of benserazide hydrochloride and to explore the molecular mechanisms involving in the anti-inflammation and anti-atherosclerosis of benserazide hydrochloride.The experiments were divided into blank groups(PBS+0.5%FBS DMEM medium),model group[LPS(500μg/mL)+0.5%FBS DMEM medium]and drug group[LPS(500μg/mL)+benserazide hydrochloride+0.5%FBS DMEM medium].Western blot,ELISA and qPCR were used to detect the protein and mRNA expression levels of inflammatory cytokines SAP,TNF-αand MCP-1 in HUVECs cells.The expression levels of p65/p-p65,p38/p-p38,IκBα/p-IκBα,AKT/p-AKT and the nuclear translocations of p65,p38 and IκBαwere detected by Western blot.The results showed that benserazide hydrochloride(1×10-9,1×10-10,1×10-11 mol/L)could significantly inhibit the protein and mRNA expression of pro-inflammatory cytokines SAP,TNF-αand MCP-1.Besides,it could down-regulate the protein expression of p65/p-p65,p38/p-p38,IκBα/p-IκBαand AKT/p-AKT in the signal pathway while inhibiting the nuclear translocation of p65,p38 and IκBα,thereby inhibiting the transcriptional activity of the related genes.