人参皂苷Re对氧化低密度脂蛋白诱导的人脐静脉内皮细胞MEK/ERK/NF-κB信号通路的影响

目的基于丝裂原活化蛋白激酶/细胞外调节蛋白激酶/核因子-κB（MEK/ERK/NF-κB）信号通路探讨人参皂苷Re（G-Re）对氧化低密度脂蛋白（ox-LDL）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，分别设置空白对照组、模型组（ox-LDL 150 μg/mL）、G-Re组（20 μmol/L）、G-Re 加 MEK抑制剂组（G-Re 20 μmol/L + U0126-EtOH 10 μmol/L）。采用CCK8法检测细胞活力；ELISA检测细胞上清液炎症因子IL-6和TNF-α表达水平；免疫荧光检测NF-κB蛋白的荧光强度；Western Blot和RT-PCR检测MEK、ERK和NF-κB蛋白和基因表达。结果与空白对照组比较，模型组细胞活力减弱（P＜0.01），炎症因子IL-6和TNF-α表达升高（P＜0.05），NF-κB蛋白的荧光强度增加（P＜0.05），MEK、ERK和NF-κB蛋白及基因表达上调（P＜0.05，P＜0.01）。与模型组比较，G-Re组干预后细胞活力增强（P＜0.05，P＜0.01），炎症因子IL-6和TNF-α表达下降（P＜0.05，P＜0.01），NF-κB蛋白的荧光强度降低（P＜0.05），MEK、ERK和NF-κB蛋白及基因表达下调（P＜0.05，P＜0.01）。与G-Re组比较，G-Re加MEK抑制剂组ERK和NF-κB基因和蛋白表达上调（P＜0.05，P＜0.01）。结论 G-Re能够减轻ox-LDL诱导的HUVECs的炎症反应和内皮损伤，其作用机制可能与抑制MEK/ERK/NF-κB信号通路的激活有关。

英文摘要:

Objective To explore the effects of Ginsenoside Re（G-Re）on oxidized low density lipoprotein（ox-LDL）-induced inflammatory response via mitogen-activated protein kinase/extracellular regulated protein kinases/nuclear factor kappa-B（MEK/ERK/NF-κB） signaling pathway in human umbilical vein endothelial cells（HUVECs）. Methods HUVECs were cultured in vitro. The experiments were divided into 4 groups， including control group， model group（ox-LDL 150 μg/mL）， ox-LDL + G-Re group（G-Re 20 μmol/L）， ox-LDL + G-Re + MEK inhibitor group（G-Re 20 μmol/L + U0126-EtOH 10 μmol/L）. Cell viability was measured by CCK-8 assay， the levles of IL-6 and TNF-α were detected with enzyme-linked immunosorbent assays（ELISA）， the protein expression of NF-κB was detected by immunofluorescence staining（IF）. The mRNA and protein expression of MEK， ERK and NF-κB were respectively measured by Real time polymerase chain reaction（RT-PCR）and Western Blot. Results Compared with the control group， cell viability declined（P＜0.01）， the levles of IL-6 and TNF-α increased（P＜0.05）， the fluorescence intensity of NF-κB protein increased（P＜0.05）， the mRNA and protein expression of MEK， ERK and NF-κB increased（P＜0.05，P＜0.01）in the model group. Compared with the model group， cell viability increased（P＜0.05，P＜0.01）， the levles of IL-6， and TNF-α reduced（P＜0.05，P＜0.01）， the fluorescence intensity of NF-κB decreased（P＜0.05）， the mRNA and protein expression of MEK， ERK and NF-κB decreased（P＜0.05，P＜0.01）in the G-Re group. Compared with G-Re group，protein and mRNA expression of ERK and NF-κB decreased in ox-LDL + G-Re + MEK inhibitor group（P＜0.05，P＜0.01）. Conclusion G-Re reduced inflammatory response and endothelial injury of HUVECs induced by ox LDL， and the mechanism might be related to inhibit the activation of MEK/ERK/NF-κB signaling pathway.

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