丁烯酸内酯对心肌线粒体呼吸链酶复合物活力的影响  

Effect of butenolide on the activities of complexes Ⅰ-Ⅳ in the respiratory chain of myocardium mitochondria

作　　者：刘井波[1] 彭双清[1] 阳海鹰[1] 王国强[1] 

机构地区：[1]军事医学科学院毒物药物研究所国家北京药物安全评价研究中心,北京100850

出　　处：《毒理学杂志》2005年第1期18-20,共3页Journal of Toxicology

基　　金：国家自然科学基金 (30 340 0 0 4 )

摘　　要：目的　研究丁烯酸内酯 (BUT)对大鼠心肌细胞毒性作用机制。方法　差速贴壁结合化学方法分离纯化大鼠心肌细胞。MTT法检测BUT作用于心肌细胞后其存活率的改变。用差速离心法分离心肌线粒体。TBA法检测心肌匀浆和心肌线粒体悬液中MDA的含量。应用紫外分光光度法检测BUT作用于线粒体悬液 2h后线粒体呼吸链酶复合物活力。结果　BUT对培养的心肌细胞有很强的毒性作用并且呈现明显的量效 关系 ,其作用于心肌细胞的LC50 为5 3 μg/mL。BUT作用在线粒体悬液中所引起的脂质过氧化效应强于其在心肌匀浆中所引起的脂质过氧化作用。BUT作用后线粒体酶复合物活力发生了改变 ,复合物Ⅰ和复合物Ⅱ的活力受到了比较明显的抑制 ,而复合物Ⅲ、Ⅳ的活力则有一定程度升高。结论　BUT能引起心肌细胞的脂质过氧化作用 ,这种作用有可能是因为BUT作用于线粒体呼吸链影响了电子在呼吸链上的传递过程所致。Objective To study the mechanism of the cytotoxicity induced by butenolide (BUT) on myocyt es.Methods Cultures of myocytes were performed by differential plating method and myocyte viability was tested by applying the MTT-test.Mitoch ondria were isolated by the conventional differential centrifugation method from the hearts of Wistar rats.Malondialdehyde (MDA) was quantified by the thiobarbi turic acid (TBA) method Activities of respiratory chain complexes was determine d by an UV-spectrophotometer.Results BUT showed toxicity in a d ose-dependent manner to myocyte and the LC 50 value was about 5.3 μg/ml. The activities of complex Ⅰ and Ⅱ decreased in treated mitochondrial suspensio n with BUT when compared with controls.On the contrary,the activities of complex Ⅲ and Ⅳ increased in treated mitochondrial suspention.Conclusion Primary culture myocytes are sensitive to the toxicity of BUT.Our results indic ate that heart is one of the major organs damaged by lipid peroxidation caused b y BUT.

关 键 词：线粒体呼吸链 丁烯酸内酯 酶复合物 活力 脂质过氧化作用 紫外分光光度法 心肌细胞 心肌线粒体 毒性作用机制 MTT法检测 BUT 差速离心法 分离纯化 化学方法 TBA法 LC50 复合物Ⅰ 肌匀浆 存活率 MDA 悬液 大鼠 

分 类 号：R749.16[医药卫生—神经病学与精神病学] R684.1[医药卫生—临床医学]