【摘要】 目的:观察槲皮素对过氧化氢所致心肌细胞损伤的保护作用及其机制。方法:①实验于2004-12/2005-05在锦州医学院药理学教研室完成。选用出生1～3d的SD大鼠15只,雌雄不拘。②采用纯化培养的心肌细胞建立过氧化氢损伤模型,实验分为6组(每6孔细胞一组):正常对照组(正常细胞培养,加入等数量的培养基,不加入任何药物),10g/L二甲基亚砜组(加入10g/L二甲基亚砜),过氧化氢损伤组(加入过氧化氢200μmol/L),过氧化氢+低浓度槲皮素组(加入过氧化氢200μmol/L和槲皮素25μmol/L),过氧化氢+槲皮素中浓度组(加入过氧化氢200μmol/L和槲皮素50μmol/L),过氧化氢+槲皮素高浓度组(加入过氧化氢200μmol/L和槲皮素100μmol/L)。过氧化氢(200μmol/L)损伤时间为12h。③采用乳酸脱氢酶试剂盒测其释放量;采用黄嘌呤氧化酶法测定超氧化物歧化酶活力;硫代巴比妥酸显色法测定丙二醛含量;硝酸还原法测定一氧化氮含量;四唑盐比色法测定线粒体脱氢酶活性。④计量资料差异比较采用t检验,组间数据处理根据处理方差齐性分析结果,采用非配对t检验。结果:①心肌细胞乳酸脱氢酶活性及丙二醛含量:过氧化氢损伤组明显高于正常对照组(P<0.01),过氧化氢+高、中、低浓度槲皮素组明显低于过氧化氢损伤组(P<0.01)。②心肌细胞超氧化物歧化酶活性:过氧化氢损伤组明显低于正常对照组(P<0.01),过氧化氢+高、中浓度槲皮素组明显高于过氧化氢损伤组(P<0.01)。③线粒体脱氢酶活性:过氧化氢损伤组明显低于正常对照组(P<0.01),过氧化氢+各浓度槲皮素组明显高于过氧化氢损伤组(P<0.01)。④心肌细胞内一氧化氮含量:过氧化氢损伤组高于正常对照组(P<0.05),过氧化氢+高、中、低浓度槲皮素组低于过氧化氢损伤组(P<0.05)。结论:槲皮素具有对抗缺氧复氧损伤、明显保护心肌细胞的作用,可能与其抗氧化、抗脂质过氧化反应有关。更多还原

【Abstract】 AIM: To observe the protective effect and mechanism of quercetin on cardiomyocytes injured by hydrogen dioxide.METHODS: ①The experiment was performed in the Stuff Room of Pharmacology, Jinzhou Medical University from December 2004 to May 2005. 15 SD rats aged 1-3 days old were selected. ②Hydrogen dioxide injured model was made with purified cultured cardiomyocytes, and the experiment was divided into 6 groups with 6 pores in each group: Normal control group (normal cell was cultivated with only medium of the same volume ); 10 g/L Dimathyl sulfoxide group(10 g/L dimathyl sulfoxide); Hydrogen peroxide injured group(200 μmol/L hydrogen peroxide); Hydrogen peroxide + low concentration quercetin group(200 μmol/L hydrogen peroxide + 25 μmol/L quercetin); Hydrogen peroxide + middle concentration quercetin group (200 μmol/L hydrogen peroxide + 50 μmol/L quercetin); Hydrogen peroxide + high concentration quercetin group(200 μmol/L hydrogen peroxide + 50 μmol/L quercetin). The injuring time of hydrogen peroxide (200 μmol/L) was 12 hours.③The releasing amounts were measured with lactic dehydrogenase (LDH) method and level of superoxide dismutase (SOD) was assayed with xanthine oxidase method; The contents of malondialdehyde (MDA) were measured with thiobarbituric acid mehod; The contents of nitrogen monoxide were detected with acid isoxidation; The activities of dehydrogenase in mito-chondria were detected with tetrazolium. ④The differences of measurement data were compared with t test and data among groups were processed with non-paired t test according to the analysis of varience homogeneity .RESULTS: ①The contents of activity in LDH and MDA: It was significantly higher in the hydrogen peroxide injured group than that in the normal control group, hydrogen peroxide + low, middle, high concentration quercetin group (P < 0.01).②The activities of SOD:It was significantly lower in the hydrogen peroxide injured group than that in the hydrogen peroxide + low, middle, high concentration group(P < 0.01).③The activities of dehydrogenase in mito-chondria: It was significantly lower in the hydrogen peroxide injured group than that in the hydrogen peroxide + low, middle, high concentration group(P < 0.01). ④The contents of nitrogen monoxide in cardiomyocytes: It was obviously higher in the hydrogen peroxide injured group than that in the normal control group and hydrogen peroxide + high, middle, low concentration quercetin group(P < 0.05). CONCLUSION: Quercetin has a significant protective effect on anoxia-reoxygenation of cardiomyocytes injured by hydrogen peroxide, which may relate with its antioxidation effects.更多还原