节点文献

慢性酒精中毒性肌病中线粒体损伤研究

The Damage of Mitochondra in the Chronic Alcoholic Myopathy

【作者】 齐珊珊

【导师】 王剑锋;

【作者基本信息】 大连医科大学 , 神经病学, 2009, 硕士

【摘要】 慢性酒精中毒性肌病(Chronic alcoholic myopathy,CAM)是由于长期酗酒引起的一种肌肉病,主要临床表现为肌无力、肌痛和肌萎缩,严重影响患者生活质量。研究表明氧化-抗氧化状态失衡影响了CAM的发生和发展,活性氧(ROS)在氧化-抗氧化状态失衡方面起着至关重要的作用。线粒体是细胞中重要的细胞器,它的功能除与ATP的产生有关外,近年还发现线粒体与细胞内产生的活性氧(ROS)有关。线粒体既是ROS产生的部位又是ROS损害的主要细胞器。国内外对酒精中毒性肌病有关线粒体的研究多为ATP合成方面,而对线粒体损伤的研究还未发现报道。本文分析酒精中毒性肌病中线粒体的损伤与氧化-抗氧化状态失衡的关系,进一步地探讨该病发病机制。目的:通过建立大鼠慢性酒精中毒性肌病模型,测定慢性酒精中毒性肌病发病过程中骨骼肌组织氧化抗氧化指标,检测线粒体膜流动性,线粒体细胞色素c水平以及线粒体跨膜电位等变化,来研究线粒体在慢性酒精中毒性肌病中的损伤情况并探讨发生损伤的原因,对慢性酒精中毒性肌病的发病机制进行研究,从而为该病的临床治疗提供可靠的实验依据。方法:选健康SPF级雌,雄性SD大鼠60只,随机分成两组(对照组,实验组),每组均喂饲改良的全营养高脂饲料,观察实验前后各组大鼠重量、行为学变化。于12周后,取后肢跖肌肌肉,通过组织化学染色方法确认成模。应用分光光度计法检测肌组织中各抗氧化指标(SOD、GSH-Px、MDA)的含量,分析骨骼肌氧化-抗氧化状态。通过荧光分光光度计检测线粒体膜流动性的变化以及Western Blot检测线粒体膜蛋白细胞色素c的变化。应用流式细胞仪检测线粒体膜电位。结果:1.实验初期大鼠体重无明显差异;实验后期,实验组大鼠体重与对照组有明显差异;2.生化指标检测结果:实验组大鼠骨骼肌(跖肌)中MDA含量显著高于对照组;SOD,GSH-Px活性测定实验组低于对照组;3.线粒体膜流动性改变:实验组大鼠骨骼肌(跖肌)较对照组微粘度明显增高;4.细胞色素c的表达改变:实验组大鼠骨骼肌(跖肌)蛋白含量减少;5.线粒体膜电位改变:实验组大鼠骨骼肌(跖肌)较对照组明显降低。结论:1 .慢性酒精中毒性肌病中线粒体发生损伤,提示与ROS增多有关。ROS增多使线粒体膜流动性下降,线粒体膜上细胞色素c含量减少,线粒体膜电位降低。2.慢性酒精中毒性肌病中骨骼肌线粒体膜流动性下降,反映出膜蛋白功能受损。3.线粒体膜上细胞色素c含量减少,线粒体膜电位降低提示线粒体受到损伤,02.-生成增加。

【Abstract】 The chronic alcoholic myopath (CAM) is a kind of myopathy because of longtime alcoholic misuse.Their frequently clinical appearance was having difficulties in gait,various muscle symptoms such as cramps,local pain and reduced muscle mass.So,chronic alcoholic myopathy affects the quality of patients’life severely.It’s research is more less than alcoholic liver disease,alcoholic cardiomyopathy and alcoholic peripheral nerve disease.Some research show that oxidation and anti-oxidant disbalance affect progress of CAM.It’s important for ROS in the disbalance between oxidation and anti-oxidant.Mitochondra ia a important organelle .Its functions include production ROS except synthetic ATP. Mitochondra not only the organelle in which production ROS but also the target organ which ROS attract at.The literatare about mitochondria in the CAM is most about synthesis ATP presently in China and other countries. There is no repotr about the injuries of mitochondra in the CAM.It’s important research the ralation between the injuries of mitochondra and the disbalance between oxidation and anti-oxidant. So our investigation that the alcoholie disease should be eoncentrate on the meehanism and the pathway of the damage that to the mitochondrium.Objective: By setting up rat modele of CAM,to determin the oxidation and anti-oxidation index,and to investigate the change of mitochondral membrane fluidity、the content of cyt-c、mitochondrial menbrane potential (△Ψm),which are the correlated indexes about the damage of mitochon- drion,to approach the effect and mechanism of CAM through mitochondria pathway,thus to reveal a new view of mechanism of chronic alcoholic myopathy and to provide experiment basis for clinical therapy. Methods: 60 health,male,Sprague-Dawley rats,were separated rando- mly into two groups(control and alcohol),treated on the basis of setting up the CAM rat myopathy model,at the same time,they are feeding the same improved food with high fats and sufficient nutrients,to observe the rat body weight and the change of praxiology.At the end of 12 weeks,rats were killed,takenthe muscle of posterior limb,after separating plantaris,to prepare the frozen section of the plantaris to be confirmed to form the myopathy model by the histochemical stain.Spectrophotometer was used to measure the activities of superoxide dismutase(SOD),Glutathione peroxi- dase(GSH-Px) and the content of malondialdehyde(MDA) in the muscle specimens of each group respectively. Fluorospectrophotometer was used to measure different groups of mitochondrial membrane fluidity; was used to detect the expression of cyt-c of each group and mitochondrial menbrane potential (△Ψm) was measured by flow cytometry(FCM).Results:1. The body weight(BW) of rats shows:it was significantly lower than the control group.2. Spectrophtometer method shows:MDA in alcoholic group were significantly higher than control group,activity of SOD and GSH were lower than control group.3. Fluorospectrophotometer shows: In the CAM, mitochondrial membrane fluidity descend.4. Western blot shows: the expression of cyt-c decrease in alcoholic group. 5. Flow cytometry shows: mitochondrial menbrane potential were lower in alcoholic group than control group.Conclusions:1. The increase of ROS relates to the damage of mitochondria in the CAM .In the CAM, mitochondrial membrane fluidity descend; mitochondrial menbrane potential were lower; the expression of cyt-c decrease.2. Mitochondrial membrane fluidity descend in the CAM show that the function of mitochondrial membrane protein was damaged.3. The expression of cyt-c decrease and mitochondrial menbrane potential were lower show that mitochondra were damaged. 02.- increase in CAM.

  • 【分类号】R595.6
  • 【下载频次】121
节点文献中: 

本文链接的文献网络图示:

本文的引文网络