H2O2致体外培养动脉内皮细胞损伤及山莨菪碱保护  

Artery Endothelial Cell Injury Induced by Hydrogen Peroxide and the Protective Effect of Anisodamine

作　　者：葛磊[1] 尢家騄[3] 李康华[2] 罗正曜[3] 

机构地区：[1]中南大学湘雅医院脊柱外科,湖南长沙410008 [2]中南大学湘雅医院骨科,湖南长沙410008 [3]中南大学湘雅医学院,湖南长沙410078

出　　处：《激光生物学报》2009年第2期166-171,共6页Acta Laser Biology Sinica

摘　　要：目的：实验以过氧化氢（H2O2）损伤体外培养动脉内皮细胞（AEC），对过氧化氢介导内皮细胞损伤、凋亡的机制进行探讨，观察山莨菪碱（Ani）是否抑制H2O2介导的AEC损伤、凋亡，并探讨Ani保护损伤、凋亡的机制。方法：体外培养AEC随机分组：对照组（正常培养AEC）；H2O2损伤组（0．5mmol／L H2O2损伤12h）；Ani组（不同浓度Ani：0．05，0．1，0．2mg/mL处理45min后加0．5mmol／L H2O2损伤12h）。结果：1．低浓度H2O2（0．5mmol／L）可以损伤AEC并诱导凋亡。H2O2损伤组台盼蓝着染率及LDH释放率均高于对照组（P〈0．01）；细胞总抗氧化能力（T—AOC）下降（比对照组P〈0．01）；琼脂糖凝胶电泳发现凋亡细胞的梯状电泳条带；细胞荧光染色见凋亡形态特征。2．Ani对H2O2诱导的内皮细胞损伤具保护作用。Ani组与H2O2损伤组比较，台盼蓝着染率及LDH释放率下降；T—AOC上升；在Ani0．2mg／mL组DNA琼脂糖凝胶电泳未见凋亡细胞特征性的梯状电泳条带；荧光染色未见凋亡细胞特征。结论：1．低浓度过氧化氢使AEC总抗氧化能力明显下降，耗竭细胞抗氧化能力，可致AEC的损伤、凋亡。2．山莨菪碱能减少AEC抗氧化能力的消耗，维持抗氧化能力平衡，保护0．5mmol／L H2O2介导的AEC损伤、凋亡。Qbjective ： The present study is designed to distinguish the roles of antioxidative capacity in artery endothelial cell（AEC） injury and apoptosis induced by hydrogen peroxide（ H2O2 ） and the mechanism by which anisodamine （Ani） protects AEC injury and apoptosis induced by H2O2. Methods： Artery endothelial cells（ AECs ） were randomly divided into groups as following ： control group（ cultured AEC normally） , H2O2 group（ treated with 0.5 mmoL/L H2O2 for 12 h） , Ani group （ treated with 0.5 mmoL/L H2O2 for 12 h after the pretreatment of Ani for 45 min）. Results： The results showed that exposure to low concentration of H2O2 （0.5 mmol/L） could cause AEC injure, as indicated by higher trypan blue staining rate, more LDH release, and lower cellular total antioxidative capacity（ T-AOC ） level （P 〈 0.01 vs control group）. H2O2-induced apoptosis was indicated by DNA fragmentation, florescence microscopy. Ani protected AEC from apoptosis induced by H2O2. When AECs were treated with Ani（0. 2 mg/mL） then injured by H2O2 45 min later, the DNA Ladder pattern specific in apoptotic cells were not detected, and less apoptotic cells were found by fluorescence microscopy. The treatment of Ani also maintained ceils in an appropriate redox equilibrium. Conclusion： The results indicated that： 1. Expossure to low concentration of H2O2 might induce AEC injury and apoptosis through exhausting antioxidative capacity and breaking redox equilibrium. 2. Anisodamine could maintain appropriate redox equilibrium, lessen the expending of antioxidative capaeity and inhibit H2O2-induced injury and apoptosis of AEC.

关 键 词：动脉内皮细胞 细胞损伤 山莨菪碱 过氧化氢 抗氧化能力 

分 类 号：Q253[生物学—细胞生物学]