慢性酒精中毒性肌病中线粒体损伤研究
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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。
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.
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。
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.
引文
1. Kagawa,Y.el al.Regulation of energy metabolism in human cell in aging and diabetes:F0F1,mcDNA,UCPs and ROS.Commun,1999,266:662-676.
2. Srinivasa M, Srinivasula, Ramesh Hegde, et al. A conserved XIAP- interaction motif in caspase - 9 and Smac /DIABLO regulates caspase activity and apoptosis. Nature,2001,410(6824):112-116.
3. Terrence,G,et al.The internal structure of mitochondria. TIBS, 2000, 25:319-324.
4.刘树森.线粒体学与生物医学新前沿.科技前沿与学术评论,2000,23卷:35-41.
5. Bereiter-HahnJ Voth M. Dynamics of mitochondria in living eells: Shape changes,dislocations,fusion,and fission of mitochondria. Microsc Res Tech,1994, 27(3):198-219.
6. Sun F,Huo X,Zhai YJ,et al. Crystal Sturctue of Mitochondrial Respiratory Membrane protein ComplexⅡ.Cell,2005,121(7):1043-1057.
7. Chan CB , DeLeo D ,Joseph JW, et al . Increased uncoupling protein levels in beta cells are associated with impaired glucose stimulated insulin secretion : mechanism of action[J ] . Diabetes , 2001 , 50(6) : 1302-1310.
8. Hjentnes N , Fernstrom M, Zierath JR , et al . Regulation of UCP2 and UCP3 by muscle disuse and physical activity in subjects. Diabetologia , 1999,42 (5) : 826 - 830.
9. Kassis N ,Bernard C, Pusterla A , et al . Correlation between pancreatic is uncoupling protein 2 (UCP2) mRNA concentration and insulin status in rats. Int Exp Diabetes Res , 2000 ,1 (3) :185-193.
10. Klingenberg M, Echtay KS. Uncoupling proteins : the issues from a biochemist point of view. Biochim Biophys Acta , 2001 ,1504 (1) :128-143.
11. Li LX, Skorpen F ,Egeberg K, et al . Uncoupling protein 2 participatesin cellular defense against oxidative stress in clonal beta cells . Biochem Biophys Res Commun ,2001 ,282(1) :273-277.
12. Schrauwen P,Xia J,Bogardus C,et a1.Skeletal muscle uncoupling protein 3 expression is a determinant of energy expenditure in Pima Indians. Diabetes, 1999,48(1):146- 149.
13. Fleury C,Sanchis D.The mitochondrial uncoupling protein 2:Current status.Int J Biochem Cell Biol,1999,31 (11):1261-1278.
14. Pecqueur C,Alves-Guerra MC,Gelly C,et a1.Uncoupling protein 2,in vivodistribution,induction upon oxidative stress,and evidence for translantional regulation.Biol Chem .2001,276(12):8705-8712.
15.徐建兴.线粒体参与超氧自由基代谢[J ] .生物化学与生物物理进展, 1995 ,22 (2) :179-180.
16. Lester P , Maret GT,Xin WJ . Proceedings of the InternationalSymposium on Native Antioxidants : Molecular Mechanism and Health Effects . Illinois : AOCS ,1996, 530.
17. Forman HJ , Fridovich I. Superoxide dismutatase : a comparison of rat constants. Arch Biochem Biophys ,1973 ,158 :396-400.
18. Bielski BJH , Ravindra L , Arudi W, et al . A study of the reactivity of HOO./O-. with unsaturated fatty acids. J BiolChem ,1983 ,258 :4759 - 4761.
19. Laura B , Valdez , Silvia A , Silvia Lores A , et al . Reactions of peroxynitrite in the mitochondrial matrix. Free Radical Biology &Medicine ,2000 , 29(3P4) :349- 356.
20. Liu,S S and Huang.J.P.Coexistenee of a“Botetive Oxygen Cycle”with Q-Cycle in the Respiratory cliain,Molecular mechanism and health effects.AOCS Press . 1996,513-529.
21. Bonout D , Garrido A , Suazo M, et al . Effects of suppl ementation with folic acid and antioxidant vitamins on homocysteine levels and LDL oxidation in coronary patient s. Nut rition , 2000 ,16 ( 2) :107-110.
22. Junghans A , Sies H , Stahl W. Carotenoid - containing unilamellarliposomes loaded with glutathione : a model to study hydrophobi c -hydrophilic antioxidant interaction. Free Radic Res , 2001 , 33(6) :801-808.
23. Lifshitz J,Mclntosh TK.Age-saaoiciated mitochondrial DNA deletions are not evident chronically after experimental brain injury in the rat .J Neurotrauma , 2003,20(2):139-149.
24. Wallace DC , Diseases of the mtDNA. A nn Rev Biochem , 1992 (61) : 1 175～1 212.
25. Berdanier CD , Everts HB. Mitochondrial DNA in aging and degenerative disease. Mutat Res , 2001 (475) : 169-184.
26. Jacobi FK, Meyer J , Pusch CM , et al . Quantitation of heteroplasmy in mitochondrial DNA mutations by primer extension using Vent (R) (exo2) DNA polymerase and RFL P analysis. MutatRes , 2001 (478) : 141-151.
27. Kae M,Sakura N,Ueda H ,et al.Clinical implications of duplicated mitochondrialDNA in Pearson Syndome. A m J MedGenet , 2000 , 67 (6) : 205- 209.
28. Lee HC , Yin PH , Yu TN , et al . Accumulation of mitochondrial DNA deletions in human oral tissues -effects of betel quid chewing and oral cancer. Mutat Res , 2001 (493) : 67-74.
29. Namura S,Zhu J,Fink K et al.Activation and clevaage of caspase-3 in apoptosis induced by experimental cerebral ischemia.J Neurosic,1998,18:3659-3668.
30. Fujita R,Ueda H.Protein kinaseC-mediated cell death mode switch induced by high glucose.Cell Death Differ ,2003,10:1336-1347.
31. Pulsinelli WA,Jaeewiez M.Isehemic brain injury and the Therapeutic window. Ann N Y Acad Sci,1997,835:187-193.
32. Zima, T., Kalousova, M., 2005. Oxidative stress and signal transduction path- ways in alcoholic liver disease. Alcoholism, Clinical and Experimental Research 29 (11 Suppl), 110-115.
33. Lin SC, Chung CY, Chiang CL, et al. The influence of p ropolis ethanol extract on liver microsomal enzymes and glutathione after chronic alcohol administra- tion. Am J ChinMed, 1999, 27: 83293.
34. Anderson MF,Sims NR.Mitochondrial respiratory function and cell death in focal cerebral ischemia.J Neuorehem,1999,73(3):1189-1199
35. Mansouri A. An alcoholic binge causes massive degration of hepatic mitoc- hondrial DNA in mice. Gastroenterology, 2003, 121: 181-190.
36. Abdellah M, Christ D, Sabine A,et al. Acute Ethanol Administration Oxidatively Damages and Depletes Mitochondrial DNA in Mouse Liver, Brain, Heart, and Skeletal Muscles: Protective Effects of Antioxidants . Printed in U.S.A., 2001, 298 (2):737-743.
1. Preedy VR,Junko Adachi,Timothy J,et al.Recent advances in the pathology of alcoholic myopathy.Alcoholism:Clinical and Experimental Research, 2001, 25: 54-59.
2. Preedy VR,Ohlendieck K,et al.The importance of alcohol-induced muscle disease [J]. J Muscle Res Cell Motil,2003,24(1):55-63.
3. Preedy VR,Jonathan R,et al.Alcoholic muscle disease:features and mechanisms [J].Journal of pathology,1994,173:309-315.
4. Charles H,Robert A,Andrew D.Molecular mechanisms responsible for alcohol- induced myopathy in skeletal muscle and heart.The International Journal of Biochemistry&Cell Biology,2005,10:2180-2195.
5. Chance B.Sies H , Bovets A .Hydroperoxide metabolism in mammalian organs . Physiological,1979,59(3):527-605.
6.刘丽君,彭建新,洪华珠,等.线粒体在细胞凋亡中的变化与作用[ J ].细胞生物学杂志, 2005, 27 (2) : 117 - 120
7. Kagnwa, Y, et al. Regulation of energy metabolism in human cells in aging and diabetes:F0F1,mcDNA,UCPs and ROS. Biochem Bioghys Reacher . 1999 , 266: 662-676.
8. Blazovicsa,Brind AM.Oxidative stress and alcoholicl iver disease.[J] Orv Hetil, 2004,145(38):1937-1942.
9. Charriaut-marlangue C,Magraill l,RePersa A,et al. Apoptosis and necorsis atfer reversible focal isehemia: an in situ DNA frangmentation analysis. [J]Cere Blood Flow Metab,1996,16(2):186-194.
10. Xu J X, Li X,Zhang Y X,et al, Mitcchondrial respiratory chain:A Self-defense system againstoxygen toxicity Proceedings of the international symposium on natural antioxidauts:molecular mechanisms and health effects.Champaing, 1996, 530-539.
11. Li LX, Skorpen F ,Egeberg K, et al . Uncoupling protein 2 participatesin cellular defense against oxidative stress in clonal beta cells . Biochem Biophys Res Commun ,2001 ,282(1) :273-277.
12. Preedy VR,Junko Adachi,Migiwa Asano,et al.Free radicals in alcoholic myopathy:indices of damage and preventive studies.Free Radical Biology& Medicine,2002,32:683-687.
13. Kabuto H,Hasuike S,Minagawa N,&Shishibori T.Effects of bisphenol A on the metabolisms of active oxygen species in mouse tissues. Environmental Research,2003,93,31-35.
14. Veerappan RM,Senthil S,Rao MR,Ravikumar R,&Pugalendi K.V.Redox status and lipid peroxidation in alcoholic hypertensive patients and alcoholic hypertensive patients with diabetes.Clinica Chimica Acta, 2004,340,207-212.
15. Yen TC,King KL,Lee HC,et al.Age-dependent increase of mitochondrial DNA deletions together with lipid peroxides and superoxide dismutase in human liver mitochodria.Free Radical Biology and Medicine, 1994, 16(2):207.
16. Shaw S,Rubin KP,Lieberi-CS.Depressed hepatic glutathione and incressed diene conjugate in alcoholic liver disease:Evidence of lipid peroxidation.Dig Dis Sci,1983,28:585.
17. Stark G.Functional consequences of oxidative membrane damage.J Membr Biol, 2005,205:1-16.
18. Ischiropoulos H , Beckman JS. Oxidative stress and nitration in neurodegenera- tion : cause , effect , or association [J ]. J Clin Invest , 2003 ,111 (2) :163 - 169.
19. Santos JH. Cell sorting experiments link persistent mitochondrial DNA damage with loss of mitochondrial membrane potential and apoptotic cell death[J ] . J Biol Chem , 2003 , 278 (3) :1728– 1734.
20. Ji YB, Gao SY, Kong Q. Effect of sea-algae polysaccharide totumor cell membrane fluidity[J].Chin Trad Herbal drugs, 2001,33(5):435.
21.杨福愉.生物膜流动性.生物化学与生物物理学报,1984,16:223.26.
22. Iverson SL , Orrenius S. The cardiolipin-cytochrome c interaction and the mitochondrial regulation of apoptosi s [J ] . Arch Biochem Biophys , 2004 , 423 (1) :37 - 46.
23. Skuisehev V P. Cytochrome c in the apoptotic and antioxidant cascades. FEBS Lett,1998,423 (2):275-280.
24. Yusuke F,Noriko N, Yutaka F, et al. Apaf-1 Is a mediator of E2F-1-induced Apoptosis[ J ]. The Journal of Biological Chemistry, 2002, 277 (42) : 39760 - 39768.
25. Green DR, Kroemer G. The pathophysiology of mitochondrial cell death [J ] . Science , 2004 ,305 (5684) :626 - 629.
26. Zamzami N, Susin SA, Marehetti P, et al, Mitoehondrial colltrol of nuelear apoptosis[J].ExpMed,1996, 183(4):1533-1544.
27. Lieber CS.Alcolhol and the liver:1994 update. Gastroeerology, 1994, 106: 1085- 1105.
28. Crompton M. Mitoehondrial intermembrane junctional complexes and their rolein cell death[J]. JPhysiol,2000,529(l):11-21.
29. Zima, T., Kalousova, M., 2005. Oxidative stress and signal transduction pathways in alcoholic liver disease. Alcoholism, Clinical and Experimental Research 29 (11 Suppl), 110-115.
30. Spach, P.I., Bottenus, R.E., Cunningham, C.C., 1982. Control of adenine nucleotide metabolism in hepatic mitochondria from rats with ethanolinduced fatty liver. The Biochemical Journal 202, 445-452.
31. Capaldi RA. Arrangement of protein in the mitochondria inner membrane. Biochim Biophys Acta , 1982 , 694 : 291 - 295.
32. Lieber CS. Alcoholic fatty liver: its pathogenesis and mechanism of progression to inflammation and fibrosis,Aleohol,2004,34(l):9-19
33. Ly JD,Grubb DR,Lawen A.The mitochondrial membrane potential(△Ψm)in apoptosis:an update [J].Apoptosis,2003,8(2):l15-128.
34. Kroemer G, Zamzam i N,Susin SA. M itochondrial control of apopto sis. Immuno logy Today, 1997; 18 (1) : 44.
35. White RJ,Reynolds IJ.Mitochondrial depolarization inglutamate-stimulated neurons:an early signal specific to excitotoxin exposure [J].J Neurosci,1996,1 6(1 8):5688-5697
36. Glibert ME,Lasley SM.Chronic developmental lead exposure and hippocampal longterm potentiation :biphasic doseresponse relationship. Neurotoxicology, 1999,20:71-82.
37.井波,彭双清.脂质过氧化作用与线粒体损伤中国预防医学杂志2005, 6(2): 16-17.
38.蔡循,陈国强,陈竺等.线粒体跨膜电位与细胞凋亡.「J]生物化学与生物物理进展,2001,28(l)3-6.
39. Fataccioli V. Effects of chronic ethanol administration on rat liver proteasome activities: relationship with oxidative stress [ J ]. Hepatology , 2004, 34: 14220.
40. Wallace DC , Diseases of the mtDNA[J ] . A nn Rev Biochem , 1992 (61) : 1175-1212.
41. Thyagarajan B , Padua RA , Campbell C , Mammalian mitochondria possess homologous DNA recombination activity[J ] . J Biol Chem , 1996 , 271 (44) : 27 536-27 543.
42. Mastaglia FL,Walton J.Skeletal muscle pathology.Churchill Livingstone,New York,1982,1:12-1.
2. Srinivasa M, Srinivasula, Ramesh Hegde, et al. A conserved XIAP- interaction motif in caspase - 9 and Smac /DIABLO regulates caspase activity and apoptosis. Nature,2001,410(6824):112-116.
3. Terrence,G,et al.The internal structure of mitochondria. TIBS, 2000, 25:319-324.
4.刘树森.线粒体学与生物医学新前沿.科技前沿与学术评论,2000,23卷:35-41.
5. Bereiter-HahnJ Voth M. Dynamics of mitochondria in living eells: Shape changes,dislocations,fusion,and fission of mitochondria. Microsc Res Tech,1994, 27(3):198-219.
6. Sun F,Huo X,Zhai YJ,et al. Crystal Sturctue of Mitochondrial Respiratory Membrane protein ComplexⅡ.Cell,2005,121(7):1043-1057.
7. Chan CB , DeLeo D ,Joseph JW, et al . Increased uncoupling protein levels in beta cells are associated with impaired glucose stimulated insulin secretion : mechanism of action[J ] . Diabetes , 2001 , 50(6) : 1302-1310.
8. Hjentnes N , Fernstrom M, Zierath JR , et al . Regulation of UCP2 and UCP3 by muscle disuse and physical activity in subjects. Diabetologia , 1999,42 (5) : 826 - 830.
9. Kassis N ,Bernard C, Pusterla A , et al . Correlation between pancreatic is uncoupling protein 2 (UCP2) mRNA concentration and insulin status in rats. Int Exp Diabetes Res , 2000 ,1 (3) :185-193.
10. Klingenberg M, Echtay KS. Uncoupling proteins : the issues from a biochemist point of view. Biochim Biophys Acta , 2001 ,1504 (1) :128-143.
11. Li LX, Skorpen F ,Egeberg K, et al . Uncoupling protein 2 participatesin cellular defense against oxidative stress in clonal beta cells . Biochem Biophys Res Commun ,2001 ,282(1) :273-277.
12. Schrauwen P,Xia J,Bogardus C,et a1.Skeletal muscle uncoupling protein 3 expression is a determinant of energy expenditure in Pima Indians. Diabetes, 1999,48(1):146- 149.
13. Fleury C,Sanchis D.The mitochondrial uncoupling protein 2:Current status.Int J Biochem Cell Biol,1999,31 (11):1261-1278.
14. Pecqueur C,Alves-Guerra MC,Gelly C,et a1.Uncoupling protein 2,in vivodistribution,induction upon oxidative stress,and evidence for translantional regulation.Biol Chem .2001,276(12):8705-8712.
15.徐建兴.线粒体参与超氧自由基代谢[J ] .生物化学与生物物理进展, 1995 ,22 (2) :179-180.
16. Lester P , Maret GT,Xin WJ . Proceedings of the InternationalSymposium on Native Antioxidants : Molecular Mechanism and Health Effects . Illinois : AOCS ,1996, 530.
17. Forman HJ , Fridovich I. Superoxide dismutatase : a comparison of rat constants. Arch Biochem Biophys ,1973 ,158 :396-400.
18. Bielski BJH , Ravindra L , Arudi W, et al . A study of the reactivity of HOO./O-. with unsaturated fatty acids. J BiolChem ,1983 ,258 :4759 - 4761.
19. Laura B , Valdez , Silvia A , Silvia Lores A , et al . Reactions of peroxynitrite in the mitochondrial matrix. Free Radical Biology &Medicine ,2000 , 29(3P4) :349- 356.
20. Liu,S S and Huang.J.P.Coexistenee of a“Botetive Oxygen Cycle”with Q-Cycle in the Respiratory cliain,Molecular mechanism and health effects.AOCS Press . 1996,513-529.
21. Bonout D , Garrido A , Suazo M, et al . Effects of suppl ementation with folic acid and antioxidant vitamins on homocysteine levels and LDL oxidation in coronary patient s. Nut rition , 2000 ,16 ( 2) :107-110.
22. Junghans A , Sies H , Stahl W. Carotenoid - containing unilamellarliposomes loaded with glutathione : a model to study hydrophobi c -hydrophilic antioxidant interaction. Free Radic Res , 2001 , 33(6) :801-808.
23. Lifshitz J,Mclntosh TK.Age-saaoiciated mitochondrial DNA deletions are not evident chronically after experimental brain injury in the rat .J Neurotrauma , 2003,20(2):139-149.
24. Wallace DC , Diseases of the mtDNA. A nn Rev Biochem , 1992 (61) : 1 175～1 212.
25. Berdanier CD , Everts HB. Mitochondrial DNA in aging and degenerative disease. Mutat Res , 2001 (475) : 169-184.
26. Jacobi FK, Meyer J , Pusch CM , et al . Quantitation of heteroplasmy in mitochondrial DNA mutations by primer extension using Vent (R) (exo2) DNA polymerase and RFL P analysis. MutatRes , 2001 (478) : 141-151.
27. Kae M,Sakura N,Ueda H ,et al.Clinical implications of duplicated mitochondrialDNA in Pearson Syndome. A m J MedGenet , 2000 , 67 (6) : 205- 209.
28. Lee HC , Yin PH , Yu TN , et al . Accumulation of mitochondrial DNA deletions in human oral tissues -effects of betel quid chewing and oral cancer. Mutat Res , 2001 (493) : 67-74.
29. Namura S,Zhu J,Fink K et al.Activation and clevaage of caspase-3 in apoptosis induced by experimental cerebral ischemia.J Neurosic,1998,18:3659-3668.
30. Fujita R,Ueda H.Protein kinaseC-mediated cell death mode switch induced by high glucose.Cell Death Differ ,2003,10:1336-1347.
31. Pulsinelli WA,Jaeewiez M.Isehemic brain injury and the Therapeutic window. Ann N Y Acad Sci,1997,835:187-193.
32. Zima, T., Kalousova, M., 2005. Oxidative stress and signal transduction path- ways in alcoholic liver disease. Alcoholism, Clinical and Experimental Research 29 (11 Suppl), 110-115.
33. Lin SC, Chung CY, Chiang CL, et al. The influence of p ropolis ethanol extract on liver microsomal enzymes and glutathione after chronic alcohol administra- tion. Am J ChinMed, 1999, 27: 83293.
34. Anderson MF,Sims NR.Mitochondrial respiratory function and cell death in focal cerebral ischemia.J Neuorehem,1999,73(3):1189-1199
35. Mansouri A. An alcoholic binge causes massive degration of hepatic mitoc- hondrial DNA in mice. Gastroenterology, 2003, 121: 181-190.
36. Abdellah M, Christ D, Sabine A,et al. Acute Ethanol Administration Oxidatively Damages and Depletes Mitochondrial DNA in Mouse Liver, Brain, Heart, and Skeletal Muscles: Protective Effects of Antioxidants . Printed in U.S.A., 2001, 298 (2):737-743.
1. Preedy VR,Junko Adachi,Timothy J,et al.Recent advances in the pathology of alcoholic myopathy.Alcoholism:Clinical and Experimental Research, 2001, 25: 54-59.
2. Preedy VR,Ohlendieck K,et al.The importance of alcohol-induced muscle disease [J]. J Muscle Res Cell Motil,2003,24(1):55-63.
3. Preedy VR,Jonathan R,et al.Alcoholic muscle disease:features and mechanisms [J].Journal of pathology,1994,173:309-315.
4. Charles H,Robert A,Andrew D.Molecular mechanisms responsible for alcohol- induced myopathy in skeletal muscle and heart.The International Journal of Biochemistry&Cell Biology,2005,10:2180-2195.
5. Chance B.Sies H , Bovets A .Hydroperoxide metabolism in mammalian organs . Physiological,1979,59(3):527-605.
6.刘丽君,彭建新,洪华珠,等.线粒体在细胞凋亡中的变化与作用[ J ].细胞生物学杂志, 2005, 27 (2) : 117 - 120
7. Kagnwa, Y, et al. Regulation of energy metabolism in human cells in aging and diabetes:F0F1,mcDNA,UCPs and ROS. Biochem Bioghys Reacher . 1999 , 266: 662-676.
8. Blazovicsa,Brind AM.Oxidative stress and alcoholicl iver disease.[J] Orv Hetil, 2004,145(38):1937-1942.
9. Charriaut-marlangue C,Magraill l,RePersa A,et al. Apoptosis and necorsis atfer reversible focal isehemia: an in situ DNA frangmentation analysis. [J]Cere Blood Flow Metab,1996,16(2):186-194.
10. Xu J X, Li X,Zhang Y X,et al, Mitcchondrial respiratory chain:A Self-defense system againstoxygen toxicity Proceedings of the international symposium on natural antioxidauts:molecular mechanisms and health effects.Champaing, 1996, 530-539.
11. Li LX, Skorpen F ,Egeberg K, et al . Uncoupling protein 2 participatesin cellular defense against oxidative stress in clonal beta cells . Biochem Biophys Res Commun ,2001 ,282(1) :273-277.
12. Preedy VR,Junko Adachi,Migiwa Asano,et al.Free radicals in alcoholic myopathy:indices of damage and preventive studies.Free Radical Biology& Medicine,2002,32:683-687.
13. Kabuto H,Hasuike S,Minagawa N,&Shishibori T.Effects of bisphenol A on the metabolisms of active oxygen species in mouse tissues. Environmental Research,2003,93,31-35.
14. Veerappan RM,Senthil S,Rao MR,Ravikumar R,&Pugalendi K.V.Redox status and lipid peroxidation in alcoholic hypertensive patients and alcoholic hypertensive patients with diabetes.Clinica Chimica Acta, 2004,340,207-212.
15. Yen TC,King KL,Lee HC,et al.Age-dependent increase of mitochondrial DNA deletions together with lipid peroxides and superoxide dismutase in human liver mitochodria.Free Radical Biology and Medicine, 1994, 16(2):207.
16. Shaw S,Rubin KP,Lieberi-CS.Depressed hepatic glutathione and incressed diene conjugate in alcoholic liver disease:Evidence of lipid peroxidation.Dig Dis Sci,1983,28:585.
17. Stark G.Functional consequences of oxidative membrane damage.J Membr Biol, 2005,205:1-16.
18. Ischiropoulos H , Beckman JS. Oxidative stress and nitration in neurodegenera- tion : cause , effect , or association [J ]. J Clin Invest , 2003 ,111 (2) :163 - 169.
19. Santos JH. Cell sorting experiments link persistent mitochondrial DNA damage with loss of mitochondrial membrane potential and apoptotic cell death[J ] . J Biol Chem , 2003 , 278 (3) :1728– 1734.
20. Ji YB, Gao SY, Kong Q. Effect of sea-algae polysaccharide totumor cell membrane fluidity[J].Chin Trad Herbal drugs, 2001,33(5):435.
21.杨福愉.生物膜流动性.生物化学与生物物理学报,1984,16:223.26.
22. Iverson SL , Orrenius S. The cardiolipin-cytochrome c interaction and the mitochondrial regulation of apoptosi s [J ] . Arch Biochem Biophys , 2004 , 423 (1) :37 - 46.
23. Skuisehev V P. Cytochrome c in the apoptotic and antioxidant cascades. FEBS Lett,1998,423 (2):275-280.
24. Yusuke F,Noriko N, Yutaka F, et al. Apaf-1 Is a mediator of E2F-1-induced Apoptosis[ J ]. The Journal of Biological Chemistry, 2002, 277 (42) : 39760 - 39768.
25. Green DR, Kroemer G. The pathophysiology of mitochondrial cell death [J ] . Science , 2004 ,305 (5684) :626 - 629.
26. Zamzami N, Susin SA, Marehetti P, et al, Mitoehondrial colltrol of nuelear apoptosis[J].ExpMed,1996, 183(4):1533-1544.
27. Lieber CS.Alcolhol and the liver:1994 update. Gastroeerology, 1994, 106: 1085- 1105.
28. Crompton M. Mitoehondrial intermembrane junctional complexes and their rolein cell death[J]. JPhysiol,2000,529(l):11-21.
29. Zima, T., Kalousova, M., 2005. Oxidative stress and signal transduction pathways in alcoholic liver disease. Alcoholism, Clinical and Experimental Research 29 (11 Suppl), 110-115.
30. Spach, P.I., Bottenus, R.E., Cunningham, C.C., 1982. Control of adenine nucleotide metabolism in hepatic mitochondria from rats with ethanolinduced fatty liver. The Biochemical Journal 202, 445-452.
31. Capaldi RA. Arrangement of protein in the mitochondria inner membrane. Biochim Biophys Acta , 1982 , 694 : 291 - 295.
32. Lieber CS. Alcoholic fatty liver: its pathogenesis and mechanism of progression to inflammation and fibrosis,Aleohol,2004,34(l):9-19
33. Ly JD,Grubb DR,Lawen A.The mitochondrial membrane potential(△Ψm)in apoptosis:an update [J].Apoptosis,2003,8(2):l15-128.
34. Kroemer G, Zamzam i N,Susin SA. M itochondrial control of apopto sis. Immuno logy Today, 1997; 18 (1) : 44.
35. White RJ,Reynolds IJ.Mitochondrial depolarization inglutamate-stimulated neurons:an early signal specific to excitotoxin exposure [J].J Neurosci,1996,1 6(1 8):5688-5697
36. Glibert ME,Lasley SM.Chronic developmental lead exposure and hippocampal longterm potentiation :biphasic doseresponse relationship. Neurotoxicology, 1999,20:71-82.
37.井波,彭双清.脂质过氧化作用与线粒体损伤中国预防医学杂志2005, 6(2): 16-17.
38.蔡循,陈国强,陈竺等.线粒体跨膜电位与细胞凋亡.「J]生物化学与生物物理进展,2001,28(l)3-6.
39. Fataccioli V. Effects of chronic ethanol administration on rat liver proteasome activities: relationship with oxidative stress [ J ]. Hepatology , 2004, 34: 14220.
40. Wallace DC , Diseases of the mtDNA[J ] . A nn Rev Biochem , 1992 (61) : 1175-1212.
41. Thyagarajan B , Padua RA , Campbell C , Mammalian mitochondria possess homologous DNA recombination activity[J ] . J Biol Chem , 1996 , 271 (44) : 27 536-27 543.
42. Mastaglia FL,Walton J.Skeletal muscle pathology.Churchill Livingstone,New York,1982,1:12-1.