动脉粥样硬化病人血清APOC3检测及其对TNF-α和JAM-1表达影响

MEASUREMENT OF APOC3 IN THE SERUM OF ATHEROSCLEROTIC PATIENTS AND ITS EFFECT ON THE EXPRESSION OF TNF-αAND JAM-1

目的探讨载脂蛋白C3(APOC3)在动脉粥样硬化病人血清中的表达及对肿瘤坏死因子-α(TNF-α)和连接黏附分子-1(JAM-1)表达影响。方法收集24例冠状动脉无狭窄和56例冠状动脉有狭窄心脏病病人外周血清,采用实时荧光定量PCR(qRT-PCR)和蛋白质印迹法(Western blot)检测APOC3基因和蛋白表达水平。体外培养人血管内皮细胞,用不同浓度的APOC3或TNF-α作用后,应用qRT-PCR和Western blot检测TNF-α和(或)JAM-1基因和蛋白表达水平。结果APOC3在冠状动脉有狭窄心脏病病人外周血清中的表达水平为(7.42±1.68)g/L,明显高于冠状动脉无狭窄病人的(9.36±3.65)g/L,差异有显著性(t=2.790,P<0.05)。APOC3作用后的人血管内皮细胞中TNF-α、JAM-1基因和蛋白表达水平明显升高(F=28.746~178.421,P<0.001)。TNF-α作用后的人血管内皮细胞中JAM-1基因和蛋白表达水平明显升高(t=8.167、15.589,P<0.001)。结论APOC3在动脉粥样硬化病人外周血清中高表达。APOC3能够促进血管内皮细胞中TNF-α、JAM-1的表达,而TNF-α能够促进血管内皮细胞中JAM-1的表达。

Objective To investigate the expression of apolipoprotein C3(APOC3)in the serum of patients with atherosclerosis and its effect on the expression of tumor necrosis factor-α(TNF-α)and junctional adhesion molecule 1(JAM-1).Me-thods Peripheral sera were collected from 24 heart disease patients without coronary artery stenosis and 56 patients with coronary artery stenosis,and quantitative real-time PCR(qRT-PCR)and Western blot were used to determine the expression levels of APOC3 mRNA and protein.Human vascular endothelial cells were cultured in vitro and treated with different concentrations of APOC3 or TNF-α;then qRT-PCR and Western blot were performed to determine the expression levels of TNF-αand/or JAM-1 mRNA and protein.Results The heart disease patients with coronary artery stenosis had a significantly higher expression level of APOC3 in peripheral serum than those without coronary artery stenosis((7.42±1.68)g/L vs(9.36±3.65)g/L;t=2.790,P<0.05).Human vascular endothelial cells had significantly increased mRNA and protein expression of TNF-αand JAM-1 after being treated with APOC3(F=28.746-178.421,P<0.001),and had significantly increased mRNA and protein expression of JAM-1 after being treated with TNF-α(t=8.167,15.589;P<0.001).Conclusion APOC3 is highly expressed in the peripheral serum of patients with atherosclerosis.APOC3 can promote the expression of TNF-αand JAM-1 in vascular endothelial cells,while TNF-αcan promote the expression of JAM-1 in vascular endothelial cells.