人脂联素基因重组腺病毒对内皮细胞丙二醛和超氧化物歧化酶水平的影响

目的 检测人脂联素基因重组腺病毒(Ad-apM1)对人脐静脉内皮细胞(HUVEC)丙二醛和超氧化物歧化酶(SOD)水平的影响,探讨Ad-apM1的抗氧化效应.方法 将Ad-apM1转染HUVEC,观察对HUVEC增殖的影响.双抗体夹心酶联免疫法(ELISA)检测细胞培养液中人脂联素蛋白水平,硫代巴比妥酸法检测丙二醛水平,黄嘌呤氧化酶法测定SOD水平.结果 Ad-apM1对HUVEC增殖无明显影响,用感染复数(MOI)为50的Ad-apM1转染HUVEC 24、48、72 h后培养液中脂联素浓度显著升高.转染AdapM1的HUVEC细胞SOD显著升高,而丙二醛显著降低(均P<0.05).100μmol/L H2O2处理HUVEC 6、12和24 h后丙二醛水平显著升高,SOD水平显著降低(均P<0.05),而转染Ad-apM1可逆转其作用(P<0.05).结论 本研究制备的Ad-apM1能有效转染HUVEC并分泌人脂联素,具有抗氧化、减轻过氧化氢对内皮细胞的氧化损伤作用.

Effects of apM1-contained recombinant adenovirus on malonaldehyde and superoxide dismutase levels in HUVEC cells

Objective To evaluate the effects of exogenous adenovirus-mediated gene transfer of human apM1 gene (Ad-apM1) on malonaldehyde (MAD) and superoxide dismutase (SOD) levels in human umbilical vein endothelial cells (HUVEC). Methods HUVEC proliferation was measured by MTT after infection by AdapM1. Adiponetin protein level in cell culture medium of HUVEC cells infected with Ad-apM1 was detected by double antibody sandwich ELISA. The levels of MAD and SOD were measured by chromatometry. Results MTT assay showed that Ad-apMl had no effect on HUVEC proliferation. Human adiponectin protein levels in cell culture medium significantly increased after HUVEC was infected with Ad-apM1 for 24, 48, and 72 h, along with decreased MAD and increased SOD ( all P<0.05 ). MAD levels markedly increased and SOD levels decreased after HUVEC were incubated with 100 μmol/L H2O2 for 6, 12, and 24 h, and these reactions were reversed by AdapM1 transfection (all P<0.05 ). Conclusions HUVEC infected with Ad-apM1 effectively secrete human adiponectin. Ad-apM1 exerts antioxidation effect and antagonizes H2O2 -induced endothelial injury.