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目的:从线粒体呼吸链角度分析中药冬虫夏草提取液对糖尿病引起的氧化损伤的保护作用。方法:实验于1998-03/2000-07在中国科学院动物研究所生物膜与膜生物国家重点实验室完成。雄性昆明小白鼠32只,随机分为4组,每组8只。正常对照组不加任何处理因素,虫草组、维生素E组和糖尿病组小鼠用四氧嘧啶建立实验性糖尿病小鼠模型,分别给予口服虫草提取液(1mL/只)和皮下注射维生素E溶液(4mg/kg),糖尿病组不用任何保护作用药物。利用比色法从线粒体水平测定抗氧化酶活性,用铁氰化钾脉冲法测定线粒体呼吸活链Ⅱ+Ⅲ电子传递与质子泵出偶联状况(H+/2e)。结果:32只小鼠均进入结果分析。①小鼠谷胱甘肽含量,谷胱甘肽过氧化物酶活性、谷胱甘肽过氧化物酶活性和丙二醛含量比值:糖尿病组低于正常对照组,虫草组、维生素E组高于糖尿病组,差异有显著性意义(P<0.05～0.001)。②丙二醛含量:糖尿病组高于正常对照组,虫草组、维生素E组低于糖尿病组(P<0.001)。③糖尿病组线粒体呼吸链复合体Ⅱ+Ⅲ电子传递与质子泵出总比值、净比值低于正常对照组,降低了24.76%,32.31%(P<0.001)。虫草组、维生素E组线粒体呼吸链复合体Ⅱ+Ⅲ电子传递与质子泵出总比值、净比值显著回升,高于糖尿病组(P<0.001)。结论:线粒体呼吸过程产生的自由基在糖尿病的肝损伤中可能具有重要的作用,其原因与电子漏增加、电子传递与质子泵出脱偶联有关;同时也证明了虫草提取液能有效的拮抗由糖尿病引起的肝线粒体的氧化损伤。
Objective: To analyze the protective effect of Cordyceps sinensis extract on diabetes-induced oxidative damage from the perspective of mitochondrial respiratory chain. METHODS: The experiment was performed at the State Key Laboratory of Biomembrane and Membrane Biosciences, Institute of Zoology, the Chinese Academy of Sciences from March 1998 to July 2000. Thirty-two male Kunming mice were randomly divided into 4 groups of 8 in each group. In the normal control group without any treatment factors, mouse models of Cordyceps sinensis, vitamin E and diabetes were established with alloxan in experimental diabetic mice, and they were given oral Cordyceps extract (1 mL/dose) and subcutaneous vitamin E solution (respectively). 4mg/kg), diabetes group without any protective effect of drugs. The colorimetric assay was used to determine the activity of antioxidative enzymes and potassium ferrocyanide pulse method was used to determine the mitochondrial respiratory chain II+III electron transport and proton pumping coupling (H+/2e). RESULTS: All 32 mice were involved in the analysis of the results. 1The ratio of glutathione content, glutathione peroxidase activity, glutathione peroxidase activity, and malondialdehyde content in mice: diabetic group was lower than normal control group, Cordyceps group, vitamin E group was high In the diabetic group, the difference was significant (P<0.05-0.001). 2 Malondialdehyde content: diabetic group was higher than normal control group, Cordyceps group, vitamin E group was lower than diabetic group (P<0.001). 3The ratio of total electron transfer and proton pumping ratio of mitochondrial respiratory chain complex II+III in diabetic group was lower than that of normal control group, which was decreased by 24.76% and 32.31% (P<0.001). In Cordyceps group and Vitamin E group, the ratios of electron transfer and proton pump out of the mitochondrial respiratory chain complexes II+III were significantly higher than those of the diabetic group (P<0.001). Conclusion: The free radicals produced by mitochondrial respiration may play an important role in the hepatic injury of diabetes. The reasons are related to the increase of electron leakage, electron transport and decoupling of proton pumping. It also proves that Cordyceps extract can antagonize effectively. Oxidative damage of the liver mitochondria caused by diabetes.