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背景:大豆异黄酮具有多种生物活性,其抗氧化作用成为近年来研究的热点。目前,对大豆异黄酮的研究多注重于动物实验和临床观察,缺少人体细胞水平的研究。目的:观察大豆异黄酮对氧化损伤血管内皮细胞的影响。设计:对照实验观察。材料:实验于2002-01/07在天津医科大学公共卫生学院中心实验室完成。实验材料包括人脐静脉血管内皮细胞株、低密度脂蛋白、大豆异黄酮及维生素E等。方法:体外培养血管内皮细胞,实验分为6组,即空白对照组、氧化损伤对照组(丙二醛含量为1μmol/L)、氧化损伤+维生素E对照组(维生素E50μmol/L)、氧化损伤+大豆异黄酮10,50,100μmol/L组。将氧化低密度脂蛋白作用于预先加入维生素E及不同浓度大豆异黄酮孵育24h的内皮细胞,继续培养24h,测定细胞外及细胞内各抗氧化指标。主要观察指标:各组内皮细胞内外丙二醛含量、超氧化物歧化酶、谷胱甘肽过氧化物酶活性、乳酸脱氢酶释放情况及一氧化氮生成量。结果:①各组内皮细胞丙二醛含量、超氧化物歧化酶及谷胱甘肽过氧化物酶活力的比较:氧化损伤对照组丙二醛含量显著高于空白对照组(P<0.01),而超氧化物歧化酶活性和谷胱甘肽过氧化物酶活性均显著低于空白对照组(P<0.01);氧化损伤+维生素E对照组、氧化损伤+大豆异黄酮10,50,100μmol/L组的丙二醛含量显著低于氧化损伤对照组(P<0.01),而超氧化物歧化酶和谷胱甘肽过氧化物酶活性显著高于氧化损伤对照组(P<0.01)。②各组内皮细胞乳酸脱氢酶释放情况及一氧化氮生成量的比较:氧化损伤对照组乳酸脱氢酶释放百分比明显高于空白对照组(P<0.01),产生的一氧化氮含量明显低于空白对照组(P<0.01);氧化损伤+维生素E对照组、氧化损伤+大豆异黄酮10,50,100μmol/L组的乳酸脱氢酶释放百分比显著低于氧化损伤对照组(P<0.01),一氧化氮的生成量显著高于氧化损伤对照组(P<0.01)。结论:大豆异黄酮可以减轻氧化低密度脂蛋白对血管内皮细胞的氧化损伤,可能通过丙二醛含量、超氧化物歧化酶、谷胱甘肽过氧化物酶活性、乳酸脱氢酶释放情况及一氧化氮生成量等抗氧化指标起作用。
Background: Soybean isoflavones have a variety of biological activities, and their antioxidation has become a hot topic in recent years. At present, research on soybean isoflavones focuses on animal experiments and clinical observations, and lacks research on human cell levels. Objective: To observe the effect of soybean isoflavones on oxidative damage to vascular endothelial cells. Design: Control experiment observation. MATERIALS: The experiment was performed at the Central Laboratory of School of Public Health, Tianjin Medical University from January 2002 to July 2007. Experimental materials include human umbilical vein endothelial cell line, low density lipoprotein, soy isoflavones, and vitamin E. METHODS: Vascular endothelial cells were cultured in vitro and divided into 6 groups: blank control group, oxidative injury control group (malondialdehyde content 1 μmol/L), oxidative damage + vitamin E control group (vitamin E 50 μmol/L), and oxidative damage. + Soybean isoflavones 10, 50, and 100 μmol/L. Oxidized low-density lipoprotein acted on endothelial cells incubated with vitamin E and different concentrations of soy isoflavones for 24 hours, and cultured for 24 hours to determine the extracellular and intracellular antioxidant indices. MAIN OUTCOME MEASURES: Malondialdehyde content, superoxide dismutase, glutathione peroxidase activity, lactate dehydrogenase release, and nitric oxide production in and out of endothelial cells in each group. RESULTS: 1Compared MDA content, superoxide dismutase and glutathione peroxidase activity of endothelial cells in each group: The content of MDA in oxidative injury control group was significantly higher than that in blank control group (P<0.01). The activities of superoxide dismutase and glutathione peroxidase were significantly lower than those in the blank control group (P<0.01); oxidative damage + vitamin E control group, oxidative damage + soybean isoflavones 10, 50, 100 μmol/L The content of malondialdehyde in the group was significantly lower than that in the oxidative injury control group (P<0.01), while the activities of superoxide dismutase and glutathione peroxidase were significantly higher than those in the oxidative injury control group (P<0.01). 2Comparison of lactate dehydrogenase release and nitric oxide production in endothelial cells of each group: The percentage of lactate dehydrogenase released from oxidative injury control group was significantly higher than that of blank control group (P<0.01), resulting in significantly lower levels of nitric oxide production. In the control group (P<0.01), the percentage of lactate dehydrogenase released from the oxidative injury+vitamin E control group, oxidative injury+soy isoflavones 10,50,100 μmol/L group was significantly lower than that of the oxidative injury control group (P<0.01). Nitric oxide production was significantly higher than that in the oxidative injury control group (P<0.01). Conclusion: Soybean isoflavones can reduce the oxidative damage of oxidized low-density lipoprotein on vascular endothelial cells, possibly through the content of malondialdehyde, superoxide dismutase, glutathione peroxidase activity, release of lactate dehydrogenase and Nitric oxide production and other antioxidant indicators work.