大豆异黄酮对过氧化氢及脂多糖诱导的血管内皮细胞损伤保护机制的实验研究  

Protective effects of soybean isoflavone on human umbilical vein endothelial cell injury induced by H2O2 and lipopolysaccharide

作　　者：王燕 补蔚萍 谢宏 曲爱华 刘军 

机构地区：[1]第四军医大学第二附属医院疼痛生物研究所,西安710038

出　　处：《中华心血管病杂志》2014年第2期150-155,共6页Chinese Journal of Cardiology

摘　　要：目的 探讨大豆异黄酮（soybean isoflavone,SI）对人脐静脉内皮细胞（HUVEC）损伤的保护作用及其机制.方法 建立内皮细胞脂质过氧化损伤模型.然后实验分为9组：（1）正常对照组；（2）过氧化氢（H2O2）氧化损伤组（H2O2组）：在培养基中加入2 mmol/L的H2O2诱导损伤4h;（3）脂多糖（LPS）氧化损伤组（LPS组）：在培养基中加入2 mmol/L的LPS诱导损伤4 h;（4）H2O2加SI低剂量（1 mg/ml）组（H2O2＋SI低剂量组）、中剂量（2.5 mg/ml）组（H2O1＋SI中剂量组）和高剂量（5 mg/ml）组（H2O2＋SI高剂量组）；（5）LPS加SI低剂量（1 mg/ml）组（LPS＋ SI低剂量组）、中剂量（2.5 mg/ml）组（LPS＋ SI低剂量组）和高剂量（5 mg/ml）组（LPS＋ SI高剂量组）.MTT法观察SI对HUVEC活性的影响.双波长荧光分光光度法测定HUVEC细胞内游离Ca2＋浓度.检测各组内皮细胞匀浆中丙二醛（MDA）含量及超氧化物歧化酶（SOD）、谷胱甘肽过氧化物酶（GSH-Px）活性.ELISA法测定培养的细胞上清液中白细胞介素6（IL-6）表达的变化.测定细胞凋亡率.结果 （1）H2O2组和LPS组较正常对照组血管内皮细胞损伤明显、细胞增殖少（P均＜0.05）、内皮细胞内游离Ca2＋浓度高（P均＜0.05）,细胞培养上清液和细胞匀浆中MDA含量和IL-6均高（P均＜0.05）,SOD、GSH-Px活性均低（P均＜0.05）,凋亡率均高,分别为（37.8±1.8）％和（38.9±1.1）％（P均＜0.05）,上述指标H2O1组和LPS组组间比较差异均无统计学意义.（2）H2O2＋SI中剂量组和LPS＋ SI中剂量组SOD和GSH-Px分别明显高于H2O2组和LPS组（P均＜0.05）,MDA含量、细胞内游离Ca2＋浓度和IL-6则分别明显低于H2O2组和LPS组（P均＜0.05）,凋亡细胞数量亦分别低于H2O2组和LPS组（P均＜0.05）.结论 SI可保护和修复H2O2或LPS引起的血管内皮细胞损伤,其机制可能与保护细胞的线粒体,提高细胞的抗氧化酶活性,降低细胞�Objective To investigate the protective effects and related mechanisms of soybean isoflavone （SI） on human umbilical vein endothelial cells （HUVECs） injury induced by H202 and lipopolysaceharide （LPS）. Methods H2O2 and LPS were used to induce HUVECs injury in vitro. Nine experimental groups were examined： control group, H2O2（2 mmoL/L for 4 h） , LPS （2 mmol/L for 4 h） , H2O2 ＋ low dose SI （1 mg/ml）, H2O2 ＋ moderate dose SI （2.5 mg/ml）, H2O2 ＋ high dose SI （5 mg/ml） , LPS ＋ low dose SI （ 1 mg/ml） , LPS ＋ moderate dose SI （2. 5 mg/ml） , LPS ＋ high dose SI （5 mg/ml）. The survival ratio of HUVECs was detected with MTT assay. The cultured cells were loaded by Fura-2/AM and the change of [ Ca2 ＋ ] in HUVECs was measured by fluorospectrophotometry. The contents of malondialdehyde （MDA） , superoxide dismutase （SOD） , reduced glutathione （GSH-Px） were measured by the commercial kits. The levels of tissue plasminogen activator IL-6 in the supematant were measured by enzyme linked immunosorbent assay （ELISA） kits. Apoptosis rate of the HUVECs was analyzed by flow cytometry. Results H2O2 and LPS significantly decreased HUVECs viability, increased the contents of MDA, IL-6 and decreased the contents of SOD and GSH-Px, and increased the apoptosis rate [ （37.8 ± 1. 8） % and （38.9 ± 1.1 ） % ]. Co-treatment with SI could reduce MDA and IL-6 while increase SOD and GSH-Px and reduce apoptosis in a dose-dependent manner. Conclusion The findings demonstrate that soybean isoflavone could attenuate H2O2 and LPS induced injury in human umbilical vein endothelial cells through protecting mitochondrial function, improving antioxygenie activity, and suppressing the mobilization of cytosolie calcium.

关 键 词：内皮细胞 异黄酮类 过氧化氢 脂多糖类 

分 类 号：R285[医药卫生—中药学]