晚期氧化蛋白产物通过p38丝裂原活化蛋白激酶通路促进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 in ECV304 Cells Through p38 Mitogen-Activated Protein Kinase Signal Pathway

作　　者：史春虹[1] 姜一农[2] 单路娟[3] 魏明丽[4] 路岩[2] 

机构地区：[1]大连医科大学附属一院内分泌科,辽宁省大连市116011 [2]大连医科大学附属一院心内科,辽宁省大连市116011 [3]大连医科大学附属一院中心实验室,辽宁省大连市116011 [4]大连医科大学附属一院干部病房,辽宁省大连市116011

出　　处：《中国动脉硬化杂志》2010年第10期761-764,共4页Chinese Journal of Arteriosclerosis

基　　金：国家自然科学基金(30271433)

摘　　要：目的观察晚期氧化蛋白产物对ECV304细胞基质细胞衍生因子1α表达的影响,并探讨其作用机制。方法牛血清白蛋白与次氯酸钠等量混合体外制备晚期氧化蛋白产物,RT-PCR法检测ECV304细胞基质细胞衍生因子1αmRNA的表达,Western Blotting法测定基质细胞衍生因子1α以及磷酸化p38丝裂原活化蛋白激酶(p-p38MAPK)蛋白水平,以p38MAPK特异性阻断剂SB203580进行阻断实验,酶联免疫吸附法测定上清液中基质细胞衍生因子1α浓度。结果与对照组比较,200μmol/L晚期氧化蛋白产物处理ECV304细胞2h后基质细胞衍生因子1αmRNA及蛋白表达量显著升高(P均<0.05);基质细胞衍生因子1αmRNA表达量6h达高峰(P<0.01),之后逐渐下降,24h时与对照组比较差异无显著性;随时间的推移基质细胞衍生因子1α蛋白表达量逐渐升高,24h表达量最高(P<0.01)。晚期氧化蛋白产物作用15min后p38丝裂原活化蛋白激酶磷酸化水平明显增强(P<0.05),不同浓度(0.1μmol/L、1μmol/L、10μmol/L)SB203580降低了晚期氧化蛋白产物刺激下基质细胞衍生因子1α蛋白的表达(分别为618.85±60.12、500.98±69.47和359.97±59.81,P<0.05或0.01)。结论晚期氧化蛋白产物可诱导ECV304细胞基质细胞衍生因子1α表达,p38MAPK通路是介导这一作用的重要途径之一。Aim To explore the effects of advanced oxidation protein products（AOPP）on expressions of stromal cell-derived factor-1α（SDF-1α）in ECV304 cells and the signal pathway that mediated the effects.Methods AOPP-BSA was made from bovine serum albumin（BSA）and sodium hypochlorite.After different time treated with AOPP-BSA,the expressions of SDF-1α mRNA in ECV304 cells were measured by reverse transcription polymerase chain reaction（RT-PCR）and the expressions of SDF-1α protein and the levels of phosphored-p38 mitogen-activated protein kinase（MAPK）were analyzed by Western Blotting.In inhibition test SB203580,the special inhibitor of p38MAPK of different concentrations were added into ECV304 culture media for 1 hour,then the cells were treated with AOPP-BSA for 12 hours,at last the protein levels in supernatant were measured by enzyme linked immunosorbent assay（ELISlA）.Results Compared with control group,the expressions of SDF-1α mRNA and protein in ECV304 cells increased significantly after incubated with 200 μmol/L AOPP-BSA for 2 hours（P〈0.05）.The expression of SDF-1α mRNA peaked after 6 hours（P〈0.01）and decreased gradually until there was no differentiation at 24 hours.Expression of SDF-1α protein increased in a time-dependent manner and peaked at 24 hours（P〈0.01）.After 15 minutes the levels of phosphored-p38MAPK increased significantly（P〈0.05）.When the p38MAPK pathway was blocked by SB203580（0.1 μmol/L,1 μmol/L,10 μmol/L）,the promoting effects of AOPP-BSA on expressions of SDF-1α protein in ECV304 cells were significantly inhibited（618.85±60.12,500.98±69.47,359.97±59.81,P〈0.05 or 0.01）.Conclusion AOPP induced the expression of SDF-1α from ECV304 cells,p38MAPK 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1Α ECV304 P38丝裂原活化蛋白激酶 

分 类 号：R363[医药卫生—病理学]