晚期氧化蛋白产物通过ERK通路促进ECV304细胞基质细胞衍生因子-1α的表达  

Advanced oxidation protein products promote expression of stromal-cell derived factor-1α of ECV304 cells through ERK signal pathway

作　　者：史春虹[1] 姜一农[2] 单路娟[3] 路岩[2] 张英[2] 高彦国[4] 

机构地区：[1]大连医科大学附属一院内分泌科,辽宁大连116011 [2]大连医科大学附属一院心内科,辽宁大连116011 [3]大连医科大学附属一院中心实验室,辽宁大连116011 [4]大连大学附属新华医院,辽宁大连116011

出　　处：《中国应用生理学杂志》2013年第2期142-146,共5页Chinese Journal of Applied Physiology

基　　金：国家自然科学基金资助项目(30271433)

摘　　要：目的:观察晚期氧化蛋白产物(AOPP)对ECV304细胞基质细胞衍生因子1α(SDF-1α)表达的影响,并探讨其作用机制。方法:牛血清白蛋白(BSA)与次氯酸钠等量混合体外制备AOPP-BSA,不同浓度AOPP-BSA作用后以RT-PCR法检测ECV304细胞SDF-1αmRNA的表达,Western-blot法测定SDF-1α以及磷酸化细胞外信号调节激酶(p-ERK)蛋白水平,阻断实验中先以不同浓度的ERK特异性阻断剂U0126与ECV304孵育1 h,然后与AOPP-BSA共同孵育24 h,酶联免疫吸附法测定上清液中SDF-1α蛋白浓度。结果:AOPP呈浓度依赖性的促进ECV304细胞SDF-1αmRNA表达及蛋白合成增加(P均<0.01),200μmol/LAOPP-BSA作用15 min后ERK1/2磷酸化水平明显增强(P<0.01),阻断ERK1/2通路显著抑制AOPP对ECV304细胞SDF-1α蛋白合成的促进作用(P<0.05)。结论:AOPP可诱导ECV304细胞SDF-1α的表达,ERK1/2通路是介导这一作用的重要途径之一。Objective： To explore the effects of advanced oxidation protein products （AOPP） on expressions of stromal cell-derived factor- la （SDF-la） in ECV304 cells and the signal pathway that mediated the effects. Metl AOPP-BSA was made from bovine senan albumin （BSA） and sodium hypoehlorite. After treated with AOPP-BSA of different coneentratiom（50, 100, 200/maol/L）, the expressions of SDF-la mRNA in ECV304 cells were measured by reverse transcripfion-polymerase chain reaction（RT-PCR） and the expressions of SDF-la protein and the levels of phosphorylated extrallular signal-regulated kinase（ERK） in ECV304 cells were analyzed by Western blot. In inhibition test, U0126, the special inhibitor of ERK of different concentrafiom（0.1, 1, 10 tnnol/L） were added into ECV304 cells culture media for 1 hour, then the cells were treated with AOPP-BSA for 24 hours, at last the protein levels in supernatant were detected by enzyme-linked immunosorbent assay . Results： A OPP-BSA obviously promoted the expressions of SDF-1 a mRNA and increased the levels of SDF-la protein of ECV304 ceils in dose-dependent manner（all P 〈 0.01）, after 15 minutes treated with 200 ttmol/L AOPP-BSA, the levels of phosphorylated ERK of ECV304 ceils increased significantly（ P 〈 0.01）. When the ERK pathway was blocked by U0126, the promoting effects of AOPP-BSA on expressions of SDF-la protein in ECV304 cells were significantly inhibited in dose-dependent manner（ P 〈 0.05）. C.otae.lutdea： AOPP induced the expression of SDF-la of ECV304 cells, ERK signal pathway is an important pathway that mediated the effects.

关 键 词：晚期氧化蛋白产物 基质细胞衍生因子let ECV304 

分 类 号：R363[医药卫生—病理学]