火麻仁油与海藻油混合物对氧化性低密度脂蛋白诱导的内皮细胞损伤的分子保护机制研究

Molecular protection mechanism of hempseed oil and algal oil mixture on endothelial cells injury induced by oxidized low density lipoprotein

目的:探讨由植物来源的多不饱和脂肪酸火麻仁油与海藻油混合而成的混合物(简称AH)对氧化性低密度脂蛋白(ox-LDL)诱导的内皮细胞损伤的保护作用。方法:用ox-LDL刺激人脐静脉内皮细胞株(HUVEC-12)建立内皮细胞损伤模型,药物干预后检测细胞内NO含量,流式细胞仪检测ROS变化,TUNEL法检测细胞凋亡,RT-PCR方法检测IL-6、TNF-α、NF-κB、pAMPK、SIRT1的基因表达水平,Western-Blot方法检测P-IκB、pAMPK、SIRT1蛋白表达水平。结果:与模型组相比,多不饱和脂肪酸混合物AH可以显著减少细胞NO合成,减少ROS生成,抑制细胞凋亡,降低IL-6、TNF-α、NF-κB水平,增加pAMPK、SIRT1水平。结论:AH能够通过降低内皮炎症因子来对抗ox-LDL诱导的内皮损伤,其作用机制可能与AMPK/SIRT1/NF-κB信号通路有关。

OBJECTIVE To investigate the protective effect of plant-derived polyunsaturated fatty acid mixture(algal oil and hempseed oil,‘AH’ for short) on oxidative low density lipoprotein-induced endothelial injury. METHODS Human umbilical vein endothelial cell line(HUVEC-12) was stimulated with ox-LDL to establish an endothelial cell injury model. After drug intervention, the intracellular NO content was measured. Changes of ROS were detected by flow cytometry along with apoptosis detected by TUNEL method. RT-PCR were used to analyze the mRNA expression level of IL-6, TNF-α, NF-κB, pAMPK and SIRT1.Western-Blot was used to analyze the protein expression level of P-IκB, pAMPK and SIRT1. RESULTS Compared with the model group, the polyunsaturated fatty acid mixture AH can reduce NO synthesis and ROS production, inhibit cell apoptosis, reduce the levels of IL-6, TNF-α and NF-κB as well as increase the expression level of pAMPK and SIRT1. CONCLUSION AH can protect against ox-LDL-induced endothelial injury by decreasing endothelial inflammatory factors, the mechanism of which may be related to the inhibition of AMPK/SIRT1/NF-κB signaling pathway.