芍药苷配伍小檗碱对TNF-α诱导人脐静脉内皮细胞NF-κB/YY1信号通路的影响

Effects of the Compatibility of Paeoniflorin Combined with Berberine on TNF-a Induced NF-k B/YY1 Signaling Pathway in HUVECs

目的基于NF-κB/YY1信号通路探讨芍药苷配伍小檗碱对肿瘤坏死因子α(TNF-α)诱导的人脐静脉内皮细胞(HUVECs)炎症反应的影响。方法体外培养HUVECs,分别设空白对照组、模型组(TNF-α20 ng/m L)、芍药苷组(PF 160μmol/L)、小檗碱组(BBR 20μmol/L)、配伍组(160μmol/L芍药苷+20μmol/L小檗碱组)。采用CCK8法检测各组细胞活力;LDH毒性实验检测细胞上清液中LDH释放量;ELISA法检测细胞上清液中IL-6和IL-8含量;RT-PCR检测NF-κB、YY1基因表达;Western Blot法检测NF-κB、YY1蛋白表达。结果与空白对照组比较,模型组细胞活力减弱,LDH漏出率增加,IL-6和IL-8含量增加,NF-κB和YY1基因及蛋白表达上调(P<0.05,P<0.01)。与模型组比较,芍药苷组、小檗碱组及配伍组干预后细胞活力增强,LDH漏出率减少,IL6和IL-8含量降低,NF-κB和YY1基因及蛋白表达下调,且芍药苷和小檗碱配伍组较单体组干预后效果更明显(P<0.05,P<0.01)。结论TNF-α可激活HUVECs NF-κB/YY1信号通路,诱导炎症反应,加重内皮损伤。芍药苷配伍小檗碱较单药使用可更有效地抑制NF-κB/YY1信号通路,减轻炎症反应进而起到血管内皮保护作用。

Objective To evaluate the effects of the compatibility of Paeoniflorin(PF)with Berberine(BBR)on inflammation of human umbilical veins endothelial cells(HUVECs)induced by TNF-αvia NF-κB/YY1 signaling pathway.Methods HUVECs were culturedin vitro and divided into control group,model group(TNF-α20 ng/m L),paeoniflorin group(PF 160μmol/L),berberine group(BBR 20μmol/L)and compatible group(PF 160μmol/L+BBR 20μmol/L).Cell viability was measured by CCK-8 assay.Lactic dehydrogenase(LDH)leakag was determined by LDH cytotoxicity assay.The concentrations of IL-6 and IL-8 were detected by ELISA.The m RNA and protein expression of NF-κB and YY1 were respectively measured by real time polymerase chain reaction(RT-PCR)and Western Blot.Results Compared with control group,cell viability was declined,while the LDH leakag and the concentrations of IL-6 and IL-8 were increased,the m RNA and protein expression of NF-κB and YY1 were up-regulated in the model group(P<0.05,P<0.01).Compared with model group,cell viability was increased,LDH leakag and the concentrations of IL-6 and IL-8 were decreased,the m RNA and protein expression of NF-κB and YY1 were down-regualated in the paenoflorin group and berberine group(P<0.05,P<0.01).Moreover,the beneficial effects of the compatible group were more obvious than the paenoflorin group or berberine group(P<0.05,P<0.01).Conclusions TNF-αcan activate NF-κB/YY1 signaling pathway of human umbilical vein endothelial cells,induce inflammatory responses,and aggravate endothelial damage.Paeoniflorin combined with berberine can more effectively inhibit NFκB/YY1 signaling pathway than single drug,reduce the inflammatory response,thus playing a role in vascular endothelial protection.