齐墩果酸对人脐静脉内皮细胞氧化损伤的保护作用  

Protective effects of oleanolic acid on oxidative damaged human umbilical vein endothelial cells

作　　者：李丙华[1] 王巧云[2] 韩志武[2] LI Bing-hua;WANG Qiao-yun;HAN Zhi-wu(Department of Pharmacy,Qingdao Eighth People's Hospital,Qingdao 266100,Shandong Province,China;Department of Pharmacy,Affiliated Hospital of Qingdao University,Qingdao 266003,Shandong Province,China)

机构地区：[1]青岛市第八人民医院药学部,山东青岛266100 [2]青岛大学附属医院药学部,山东青岛266003

出　　处：《中国临床药理学杂志》2021年第12期1548-1550,1561,共4页The Chinese Journal of Clinical Pharmacology

基　　金：国家自然科学基金资助项目(81274126)。

摘　　要：目的复制氧化性低密度脂蛋白(ox-LDL)诱导的人脐静脉内皮细胞(HUVECs)氧化损伤模型,研究齐墩果酸(OA)经激活剂蛋白-1(AP-1)/对氧磷酶-2(PON2)/活性氧(ROS)信号通路对该模型的保护作用。方法转染前,将HUVECs随机分为5组:对照组,模型组(100μg·m L-1ox-LDL),低、中、高剂量实验组(10,20,40μmol·L-1OA+100μg·m L-1ox-LDL),将AP-1抑制剂加入对照组细胞及高剂量实验组细胞中,分别命名为AP-1抑制剂组和高剂量OA+AP-1抑制剂组。将对照质粒和PON2激活质粒分别转染至HUVECs,分别命名为转染组-1和转染组-2,未经处理的细胞为对照组。蛋白质印迹法(Western Blot)检测c-jun、PON2蛋白表达;2’,7’-二氯二氢荧光素二乙酸酯(DCFH-DA)荧光探针标记法检测细胞内ROS含量。结果对照组,模型组,低、中、高剂量实验组中c-jun蛋白相对表达量分别为1.03±0.21,1.57±0.34,1.21±0.32,1.17±0.44,0.79±0.35,PON2蛋白相对表达量分别为1.12±0.27,0.62±0.37,0.74±0.33,0.88±0.35,0.94±0.42,ROS在490/525nm处相对荧光强度分别为1.00±0.22,2.22±0.25,1.76±0.32,1.57±0.36,1.49±0.28。低、中、高剂量实验组分别与对照组和模型组比较,差别均有统计学意义(P<0.05)。结论OA能够保护ox-LDL氧化损伤的HUVECs,其保护机制与降低AP-1活性、提高PON2活性、降低ROS含量有关。Objective Repeat the oxidative damage model of human umbilical vein endothelial cells(HUVECs)which is induced by oxygenized low density lipoprotein(ox-LDL),to investigate the protective effect of oleanolic acid(OA)for this model by activator protein-1(AP-1)/paraoxonase-2(PON2)/reactive oxygen species(ROS)signal pathway.Methods The HUVECs were randomly divided into five groups:control group,model group(100μg·m L-1ox-LDL)and low-/middle-/high-dose experimental group(10,20,40μmol·L-1OA+100μg·m L-1ox-LDL)before transfection.The AP-1 inhibitor was added to the control group cells and the high-dose experimental group cells,named as the AP-1 inhibitor group and the high-dose OA+AP-1 inhibitor group,respectively.The control plasmid and PON2activated plasmid were transfected into HUVECs,respectively;named as the Transfection-1 group and Transfection-2 group respectively.Untreated cells were the control group.Expression levels of c-jun and PON2 were determined by Western Blot analysis.Production of cell ROS was detected by 2’,7’-Dichlorofluorescin diacetate(DCFH-DA).Results There were statistically significant differences between the control group and the low-/middle-/high-dose experimental group in the relative expression of c-jun protein(1.03±0.21,1.57±0.34,1.21±0.32,1.17±0.44,0.79±0.35)and the relative expression of PON2protein of(1.12±0.27,0.62±0.37,0.74±0.33,0.88±0.35,0.94±0.42),the relative intensity of fluorescence at 490/525nm of ROS(1.00±0.22,2.22±0.25,1.76±0.32,1.57±0.36,1.49±0.28)(P<0.05).Conclusion OA protects ox-LDL oxidation-damaged in HUVECs.Its protective mechanism is related to the decrease of AP-1 activity,the increase of PON2 activity and the reduction of ROS content.

关 键 词：齐墩果酸 人脐静脉内皮细胞 氧化性低密度脂蛋白 活性氧 激活剂蛋白-1 抗氧化蛋白对氧磷酶-2 

分 类 号：R28[医药卫生—中药学]