Abstract:

Objective To investigate the effect of tumor necrosis factor-like weak inducer of apoptosis (TWEAK) in human umbilical vein endothelial cells (HUVECs) on lectin-like oxidized low-density lipoprotein receptor-1 (LOX-1) expression. Methods In vitro cultured HUVECs were taken intervention by final concentration of 0, 10, 50, 100 ng/ml of recombinant human TWEAK (rhTWEAK) for 24 h, and fluorescence quantitative polymerase chain reaction (PCR) technology was used to detect the relative expression of TWEAK group LOX-1 mRNA detection of different doses. Meanwhile, after HUVECs cells were intervened with by previously having added final concentration 0.2, 1, 10 μmol/L atorvastatin for 24 h, then coupled with a concentration of 100 ng/ml TWEAK incubated for 24 h, followed by PCR to detect the relative expression of LOX-1 mRNA; and enzyme-linked immunosorbent assay (ELISA) method was used to detect the cell supernatant LOX-1 protein expression. At the same time, morphological changes in each group were observed by fluorescence microscopy. Results LOX-1 mRNA 2-△△Ct values of final concentration of 0, 10, 50, 100 ng/ml TWEAK

intervention group and the relative expression of LOX-1 protein of the supernatant of each group were compared with the control group, which had statistically significant difference (P <0.001). TWEAK increased with increasing concentration of LOX-1 expression levels in different dose groups, which had significant difference (P <0.001); final concentration 0.2, 1, 10 μmol/ L atorvastatin intervention group after the intervention significantly inhibited the expression of LOX-1 mRNA and protein compared with 100 ng/ml TWEAK group, which had significant difference (P <0.001); and there were significant differences of inter-group comparison of different concentrations of atorvastatin group (P <0.001). Conclusion TWEAK can induce increased expression of human umbilical vein endothelial cells LOX-1, which may correlate with atherosclerosis. Atorvastatin could inhibit LOX-1 expression, which may be one of the mechanisms to play the anti-inflammatory and anti-atherosclerosis role.

Key words: Atherosclerosis; Tumor necrosis factor-like weak inducer of apoptosis; Lectin-like oxidized low-density lipoprotein receptor -1; Atorvastatin; Endothelial cell

摘要：

目的 探讨肿瘤坏死因子样凋亡微弱诱导剂（tumor n ecrosis f actor-like weak inducer of apoptosis， TWEAK）对人脐静脉内皮细胞（human umbilical vein endothelial cells，HUVECs）中血凝素样氧化低密度 脂蛋白受体-1（lectin-like oxidized low-density lipoprotein receptor-1，LOX-1）表达的影响。 方法 体外培养HUVECs细胞，加入终浓度为0、10、50、100 ng/ml的重组人TWEAK（recombinant human TWEAK，rhTWEAK）干预24 h，采用荧光定量聚合酶链式反应（polymerase chain reaction，PCR）技术检 测不同剂量的TWEAK组中LOX-1 mRNA的相对表达量；同时预先加入终浓度为0.2、1、10 μmol /L的 阿托伐他汀干预HUVECs细胞24 h后，再加浓度为100 ng/ml TWEAK孵育24 h，然后进行PCR检测LOX-1 mRNA的相对表达量；同时采用酶联免疫吸附测定法（enzyme linked immunosorbent assay，ELISA）检测 各组细胞上清液中LOX-1蛋白表达量。同时通过荧光显微镜观察各组细胞形态学变化。 结果 终浓度为0、10、50、100 ng/ml的TWEAK干预组LOX-1 mRNA 2﹣△△Ct值和各组细胞上清LOX-1 蛋白相对表达量与空白对照组相比，差异有统计学意义（P <0.001)。随着TWEAK浓度的增加LOX-1的 表达量增加，不同剂量组内比较，差异有显著性（P <0.001)；终浓度为0.2、1、10 μmol /L的阿托伐他 汀干预组干预以后显著抑制LOX-1 mRNA及蛋白的表达，与100 ng/ml TWEAK组相比，差异有显著性 （P <0.001），不同浓度阿托伐他汀组组内比较差异有显著性（P <0.001）。 结论 T WEAK可诱导HUVECs L OX-1的表达量增加，这可能与其具有致动脉粥样硬化作用有关；阿托 伐他汀可抑制LOX-1表达，可能是其发挥抗炎和抗动脉粥样硬化作用的机制之一。

关键词: 动脉粥样硬化; 肿瘤坏死因子样凋亡微弱诱导剂; 血凝素样氧化低密度脂蛋白受体-1; 阿托伐他汀; 内皮细胞