二氮嗪药物后适应及其机制分析
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摘要
近年来,随着临床冠脉内溶栓、PTCA 及冠脉搭桥术等治疗手段的广泛应用,心肌再灌注损伤愈来愈引起人们的广泛重视。缺血预适应是目前认为最强及最有效的内源性保护机制,一系列研究均显示缺血预适应可以显著减轻心肌缺血再灌注损伤。但是由于在临床治疗中经常遇到的是已经发生的心肌缺血,而真正能够干预的是已发生的心肌缺血阶段及再灌注阶段,因而从临床治疗的角度讲,缺血预适应的真正临床应用价值有很大的局限性。2003 年, Zhao 首先在狗的模型发现缺血后适应现象。之后,这一现象在大鼠和兔子的模型上也相继得到了证实。由于缺血后适应的干预应用于缺血发生之后和再灌注之前,具有广泛的临床应用前景,因而引起了人们的广泛关注。
缺血后适应发现后,如何建立有效的药物后适应途径,并分析其发生机制,已经提到研究日程。考虑到激活线粒体内膜KATP通道在缺血预适应中的心肌保护作用已被公认,而它的选择性激动剂二氮嗪又是一可用于临床的药物,因此我们在预实验的基础上,首先选择二氮嗪在动物建立药物后适应模型,并进一步分析在药物后适应和模拟缺血后适应中激活KATP通道产生心肌保护作用的机制,为进一步的临床研究和应用奠定基础。
一、二氮嗪药物后适应对离体大鼠心肌局部缺血/再灌注损伤的影响
目的:观察二氮嗪药物后适应,即在再灌注早期激活线粒体ATP 敏感性钾通道对缺血/再灌注心肌的保护作用。
材料与方法:利用Langendorff 逆行灌流装置对大鼠心脏离体灌流,通过对左冠状动脉前降支的结扎和松扎建立大鼠离体心脏局部缺血/再灌注模型。实验分为11 组:1 组,缺血/再灌注组。Ⅱ组,二氮嗪药物后适应组,并包括Ⅱa、Ⅱb、Ⅱc三个不同药物剂量组。Ⅲ组,pinacidil 药物后适应组。Ⅳ组,二氮嗪药物预适应组。Ⅴ组,二氮嗪加5-HD 药物后适应阻断组。Ⅵ组,二氮嗪加优降糖药物后适应组。Ⅶ组,二氮嗪加优降糖及5-HD药物后适应组。Ⅷ组,二氮嗪加atractyloside 药物后适应阻断组。Ⅸ组,
With increasing application of PTCA, CABG, intracoronary thrombolisis in acute myocardial infarction in recent years, myocardial ischemic and reperfusion injury has attracted considerable attention. Ischemic preconditioning has been proved to exert extensive protection to ischemic and reperfusion myocardium, including attenuation of myocardial cell death or apoptosis. However, as its name implies, ischemic preconditioning must be applied before an ischemic event to be protective, thus limiting its utility. It is of great encourage that Zhao et al first reported in 2003 that multiple brief periods reperfusion and ischemic intervention before reperfusion, called ischemic postconditioning, significantly reduced infarct size in dog heart model. The ischemic postconditioning phenomenon was also confirmed by Kin in rat model and Yan in rabbit model in the following research. Because ischemic postconditionong was applied after an ischemic event, it has greatly clinical appeal.
After ischemic postconditioning phenomenon was recognized, it is of great significance to establish effective pharmacological postconditioning and explore possible mechanisms underling it . Considerating that mitochondrial KATP(mito KATP) channels have a critical role in cardioprotection of ischemic preconditioning and that diazoxide, which is a selected agonist of mito KATP, can be applied to clinical treatment, the present study intends to firstly establish diazoxide pharmacological postconditioning Furthermore,we explore the mechanism. underling the diazoxide pharmacological postconditioning and mimic ischemic postconditioning medicated cardioprotective effect by activation of KATP channel, which lay strong basis for future clinical study.
Part ⅠProtective effect of diazoxide pharmocological postconditioning on myocardial regional ischemia/reperfusion injury in isolated rat heart
Objective To test whether and how diazoxide pharmacological postconditioning might protect myocardium against ischemic and reperfusion injury.
Materials and Methods An isolated myocardial regional ischemic/reperfusion rat model was established. Briefly, Wistar rat was sacrificed and heart was mounted on a Langendorff apparatus and retrogradely perfused with Tyrode’s solution. Then the isolated hearts underwent regional ischemia by ligaturing the left anterior descending (LAD) coronary artery and reperfusion by releasing the ligature. All hearts were divided into 11 groups: group Ⅰ, myocardial ischemic/reperfusion group ; group Ⅱ, diazoxide pharmacological postconditioning group , which was further devided into 3 dosage groups, Ⅱa,Ⅱb and Ⅱc groups; group Ⅲ, pinacidil pharmacological postconditioning group; group Ⅳ, diazoxide pharmacological preconditioning group; groupⅤ, diazoxide+5-HD pharmacological postconditioning group; group Ⅵ, diazoxide + glybenclamide pharmacological postconditioning group ; group Ⅶ, diazoxide + glybenclamide +5-HD pharmacological postconditioning group ; group Ⅷ, diazoxide + atractyloside pharmacological postconditioning group; group Ⅸ, diazoxide + GF 109203X pharmacological postconditioning group; group Ⅹ, PMA pharmacological postconditioning group; group Ⅺ, PMA + 5-HD postconditioning group . After completing perfusion, infarct size (NBT Staining), the leakage of CK were measured, and recovery of cardiac function at the end of reperfusion was also evaluated.
Result
(1) In groupⅠ(myocardial ischemic reperfusion group),myocardial infarct size(myocardial infarct size/total left ventricular size)was 44.4±0.9% . Compared with group Ⅰ, infarct sizes in different pharmacological postconditioning decreased to different levels: groupⅡa (diazoxide 10μmol/L
pharmacological postconditioning group) (36.2±0.7%), groupⅡb (diazoxide30 μmol/L pharmacological postconditioning group) (32.2±0.4%), group Ⅱc (diazoxide 60μmol/L pharmacological postconditioning group)( 22.2±2.1%), group Ⅲ( pinacidil pharmacological postconditioning group)( 35.2±2.2%), group Ⅹ(PMA pharmacological postconditioning group)( 31.2 ±1.9). Each of the a
缺血后适应发现后,如何建立有效的药物后适应途径,并分析其发生机制,已经提到研究日程。考虑到激活线粒体内膜KATP通道在缺血预适应中的心肌保护作用已被公认,而它的选择性激动剂二氮嗪又是一可用于临床的药物,因此我们在预实验的基础上,首先选择二氮嗪在动物建立药物后适应模型,并进一步分析在药物后适应和模拟缺血后适应中激活KATP通道产生心肌保护作用的机制,为进一步的临床研究和应用奠定基础。
一、二氮嗪药物后适应对离体大鼠心肌局部缺血/再灌注损伤的影响
目的:观察二氮嗪药物后适应,即在再灌注早期激活线粒体ATP 敏感性钾通道对缺血/再灌注心肌的保护作用。
材料与方法:利用Langendorff 逆行灌流装置对大鼠心脏离体灌流,通过对左冠状动脉前降支的结扎和松扎建立大鼠离体心脏局部缺血/再灌注模型。实验分为11 组:1 组,缺血/再灌注组。Ⅱ组,二氮嗪药物后适应组,并包括Ⅱa、Ⅱb、Ⅱc三个不同药物剂量组。Ⅲ组,pinacidil 药物后适应组。Ⅳ组,二氮嗪药物预适应组。Ⅴ组,二氮嗪加5-HD 药物后适应阻断组。Ⅵ组,二氮嗪加优降糖药物后适应组。Ⅶ组,二氮嗪加优降糖及5-HD药物后适应组。Ⅷ组,二氮嗪加atractyloside 药物后适应阻断组。Ⅸ组,
With increasing application of PTCA, CABG, intracoronary thrombolisis in acute myocardial infarction in recent years, myocardial ischemic and reperfusion injury has attracted considerable attention. Ischemic preconditioning has been proved to exert extensive protection to ischemic and reperfusion myocardium, including attenuation of myocardial cell death or apoptosis. However, as its name implies, ischemic preconditioning must be applied before an ischemic event to be protective, thus limiting its utility. It is of great encourage that Zhao et al first reported in 2003 that multiple brief periods reperfusion and ischemic intervention before reperfusion, called ischemic postconditioning, significantly reduced infarct size in dog heart model. The ischemic postconditioning phenomenon was also confirmed by Kin in rat model and Yan in rabbit model in the following research. Because ischemic postconditionong was applied after an ischemic event, it has greatly clinical appeal.
After ischemic postconditioning phenomenon was recognized, it is of great significance to establish effective pharmacological postconditioning and explore possible mechanisms underling it . Considerating that mitochondrial KATP(mito KATP) channels have a critical role in cardioprotection of ischemic preconditioning and that diazoxide, which is a selected agonist of mito KATP, can be applied to clinical treatment, the present study intends to firstly establish diazoxide pharmacological postconditioning Furthermore,we explore the mechanism. underling the diazoxide pharmacological postconditioning and mimic ischemic postconditioning medicated cardioprotective effect by activation of KATP channel, which lay strong basis for future clinical study.
Part ⅠProtective effect of diazoxide pharmocological postconditioning on myocardial regional ischemia/reperfusion injury in isolated rat heart
Objective To test whether and how diazoxide pharmacological postconditioning might protect myocardium against ischemic and reperfusion injury.
Materials and Methods An isolated myocardial regional ischemic/reperfusion rat model was established. Briefly, Wistar rat was sacrificed and heart was mounted on a Langendorff apparatus and retrogradely perfused with Tyrode’s solution. Then the isolated hearts underwent regional ischemia by ligaturing the left anterior descending (LAD) coronary artery and reperfusion by releasing the ligature. All hearts were divided into 11 groups: group Ⅰ, myocardial ischemic/reperfusion group ; group Ⅱ, diazoxide pharmacological postconditioning group , which was further devided into 3 dosage groups, Ⅱa,Ⅱb and Ⅱc groups; group Ⅲ, pinacidil pharmacological postconditioning group; group Ⅳ, diazoxide pharmacological preconditioning group; groupⅤ, diazoxide+5-HD pharmacological postconditioning group; group Ⅵ, diazoxide + glybenclamide pharmacological postconditioning group ; group Ⅶ, diazoxide + glybenclamide +5-HD pharmacological postconditioning group ; group Ⅷ, diazoxide + atractyloside pharmacological postconditioning group; group Ⅸ, diazoxide + GF 109203X pharmacological postconditioning group; group Ⅹ, PMA pharmacological postconditioning group; group Ⅺ, PMA + 5-HD postconditioning group . After completing perfusion, infarct size (NBT Staining), the leakage of CK were measured, and recovery of cardiac function at the end of reperfusion was also evaluated.
Result
(1) In groupⅠ(myocardial ischemic reperfusion group),myocardial infarct size(myocardial infarct size/total left ventricular size)was 44.4±0.9% . Compared with group Ⅰ, infarct sizes in different pharmacological postconditioning decreased to different levels: groupⅡa (diazoxide 10μmol/L
pharmacological postconditioning group) (36.2±0.7%), groupⅡb (diazoxide30 μmol/L pharmacological postconditioning group) (32.2±0.4%), group Ⅱc (diazoxide 60μmol/L pharmacological postconditioning group)( 22.2±2.1%), group Ⅲ( pinacidil pharmacological postconditioning group)( 35.2±2.2%), group Ⅹ(PMA pharmacological postconditioning group)( 31.2 ±1.9). Each of the a
引文
1.Murry CE,Jennings RB,Reimer KA.Preconditioning with ischemia ,a delay of lethal cell injury in ischemic myocardium Circulation,1986,74(5):1124-1126.
2. Liu Y ,Downey JM,Ischemic preconditioning protects against infarction in rat heart. Am, J physiol.1992:123:346-353.
3.Yellon DM.Alkhulaifi AM,Browne EE,et al. Ischaemic preconditioning limits infarct size in the rat heart Cardiovasc Res.1992:26:983-987
4. Zhao ZQ, Corvera JS, Halkos ME, et al..Inhibition of myocardial injury by ischemic postconditioning during reperfusion .Am J physiol.2003: 285:H579-H588
5.Kin H, Zhao ZQ, Sun HY, et al. Postconditioning attenuates myocardial ischemia-reperfusion injury by inhibiting events in the early minutes of reperfusion.Cardiovasc Res.2004:62:74-82.
6.Yang X-M,Downey JM,Cohen MV.Multiple,brief coronary occlusions during early reperfusion protect rabbit hearts by activation of ERK and production of nitric oxide. Circulation(Suppl)2003;108;IV745.
7.Downey JM, .Hsu MV, et al. Mitochondrial KATP channel opening during index ischemia and following myocardial reperfusion in ischemic rat hearts.J
Mol Cell Cardiol.2001:33:651-653.
8.Fryer RM, Hsu AK, Gross GJ. Et al.Mitochondrial KATP channel opening is important during index ischemia and following myocardial reperfusion in ischemic preconditioned rat hearts.J MOL Cell Cardiol 2001:33:831-834.
9.Grover GJ, Sleph PG, .Dzwonczyk S., et al .Role of myocardial ATP sensitive potassium channels in mediating prcconditioning in the dog heart and their possible interaction with adenosine A,-receptors.Circulation. 1992:86:1310-1316.
10.Schultz JEJ. Rose E. Yao Z .et al. Evidence for involvement of opioid receptors inchemic preconditioning in rat hearts .Am J Phvsiol.1995:268: H2157 -H2161
11.Wall TM. Sheehy R. Hartman JC, et al..Role of bradykinin in myocardial prcconditioning J pharmacol Exp ther.,1994:270:681-690.
12.Garlid, GJ,Paucek P,Yarov-yarovoy V, et al..Cardioprotective effect of diazoxide and its interaction with mitochondrial ATP-sensitive K+ channels: possible mechanisms of protection. Circ Res1997; 81:1072-1082.
13.Nakai Y, Horimito H, Mienos S, et al. Mitochondrial ATP-sensitive potassium channels play a dominant role in ischemic preconditioning of rabbit heart .Eur Surg Res.2001;33(2),57-63.
14.Hausenloy DJ, Maddock HL, Baxter GF ,et al.Inhibiting mitochondrial permeability transition pore opening : a new paradigm for myocardial preconditioning ?Cardivasc Res ,2002,55:534-543
15.Cross GJ, Auchampach JA. Block of the ATP-sensitive potassium channels prents myocardial preconditioning. Circ Res ,1992,70:223-233.,
16.Baker JE, Holman P, Gross GJ.Preconditioning in immature rabbit hearts:role of kATP channel. Circulation ,1999,99:1249-1254.
17.Rohman S, Weygandt H, Schelling P, et al..Involvement of ATP-sensitive potassium channels in preconditioning.. Basic Res cardial.1994;89:563-576.
18.Fryer RM ,Ellis JT, Hsu AK, et al. Ischemic preconditioning in rat :role mitochondrial ATP-sensitive K+ channels in the preservation of mitochondrial
fuction. Am J phsiol ,2000,278:H305-312.
19.Schultz JE, Qian YZ, Gross GJ, et al. The ischemia-selective K+ channels antagost,5-Hydroxydecanoate,.blocks ischemic preconditioning in the rat heart.J Mol Cell Cardial,1997:1050-1060.
20.Auchampach JA, Grover GJ.Blockade of ischemic preconditioning in dogs by the novel ATP dependent potassium channels angonist sodium . Cardivasc Res,1992,26:1054-1062.
21.Kita H, Miura T, Tsuchida A, et al. Suppression of reperfusion arrhythmias by preconditioning is inhibited by an ATP-sensitive potassium channels blocker 5-hygroxydecanoate but ,not by protein kinase C in the rat. J Cardiovasc phsrmacol,1998:32,791-797.
22.Crompton M,The mitochondrial permeability transition pone and its role in cell death .Biochem J, 1999,341:127-132
23.Griffiths EJ,Halestrap AP. Mitochondrial non-specific permeability transition pore opening during myocardial reperfusion
24.Crompton M.The mitochondrial permeability transition pone and its role in cell death .Biochem J, 1999,341:127-132
25 Halestrap A,connern C,Griffiths E,et al.Cyclosporin A binding to mitochondrial cyclophilin inhibits the permeability transition pore and protects hearts from ischemia/reperfusion injury. Mol Cell Biochem,1997,174:167-172
26.Halestrap AP, Clarke SJ, Javadov SA. Mitochondrial pores remain closed during ischemic but open upon reperfusion Biiochem J .1995;307:93-98
27. Kerr P, Suleiman M, Halstrap A, et al.Recorver of rat hearts reperfused after a period of ischemia is accompanied by reversal of the mitochondrial permeability transition and is enhanced by pyruvate . Am J physiol,1999, 276, H496-H502.
28.Argaud L ,Roesch OG, Raisky O ,et al.. Postconditioning inhibits mitochondrial permeability transition .Circulation 2005;111:194-197.
29..Fliss H, Gattinger D. Apoptosis in ischemic and reperfused rat myocardium. Circ Res,1996,79:949-956.
30.Zhao ZQ,Nakamura M,Wang NY, et al.Reperfusion induces myocardial apoptotic cell death.Cardiovas Res.2000;45:651-660
31.Gottlieb RA,Burlrson KO, Kloner RA,et al. Reperfusion injury induces apoptosis in rabbits cardiomyocytes. J Clin Ivest .1994;94:1621-1628. . Ann.N.Y. Acad.Sci., 1999,874:412-426.
32.Olivetti LG., Quaini F ,Sala R ,et al. Acute myocardial infarction in humans is associated with activation of programmed myocyte cell death in the surviving portion of the heart. J.Mol,Cell Cardiol.,28,2005-2016.
33.Saraste A.,Pulkki K, Kallajoki M, et al. Apoptosis in human acute myocatdial infarction.Circulation, 1997,95: 320-323
34.Gao F,.Gong B,Christopher TA, et al.Anti-apoptotic effect of benidipine,a long-lasting vasodilating calicium antagonist ,in ischemic/reperfused myocardial cells.Brit J pharm,2001,132:869-878
35 Piot CA, Padmanaban D,Ursell PC,et al Ischemic preconditioning decreases apoptosis in rat hearts in vivo Circulation,1997,96(6):1598.
36.Ichinose M, Yonemochi H, Sato T, et al.diazoxide triggers cardioprotection against apoptosis induced by oxidative stress. Am J Phsiol Heart Circ phsiol 2003,284:H2235-H2241.
37..Yang J ,Cortopassi G.. Induction of the mitochondrial permeability transition causes releaes of the apoptogenic factor cytochrome C .Free Radic Biol Med,1998,24:624-631.
38 Martin Crompton Mitochondrial intermembrane junctional complexes and their role in cell death J physiol .2000,529:11-21
39.Holmuhamedov EL, Ozcan C, Jahangir A, et al. Restoration of Ca2+ inhibited oxidative phosphorylation in cardiac mitochondria by mitochondrial Ca2+ unload. Mol cell biochem ,2001,220:135-140.
40.Ylitalo K, Ala-Rami A, Liimatta E, et al. Intracellur free calcium and mitochondrial membrane potential in ischemia/reperfusion and preconditioning. J Mol Cell Cardial 2000, 32:1223-1238.
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44.Gao WD, Atar D, Liu Y, ET AL.Role of troponin I proteolysis in the pathogenesis of stunned myocsdium.Circ RES.1997;80:393-399.
45.Sun HY, Wang NP, Kerendi F, et al..Hypoxic postconditioning reduced cardiomyocyte loss by inhibiting ROS generation and intracellur Ca2+ overload. Am J Physiol Heart Circ Physiol ,2005,288:H1900-H1908.
46.Dos Santos P, Kowaltowski AJ, Laclau MN, et al.Mechanisms by which opening the mitochondrial ATP-sensitive K+ channels protects the ischemic heart. Am J Phsiol Heart Circ phsiol 2002,283:H284-H295.
47. Holmuhamedov,EL,Wang L, Terzic A.ATP-sensitive K+ Channels modulate cardiac mitochondrial function. Am J Physiol Heart Circ Physiol ,1998,275: H1567-H1576..
48.Szewezyk A.The ATP-regulated K+ Channels in mitochondrial:five years after its discovery.Acta Biochim pol ,1996,43:713-719
49..McPhetson CD,Pierce GN, Cole WC. Ischemic cardioprotection by ATP-sensitive K+ channels involves high-energyphosphate preservation preservation preservation Am J Physiol Heart Cire Physiol ,1993,265: H1809-H1818.
50.Nishida M,Borzak S,Kraemer M,et al.Role of cation gradients in hypercontracture of myocytes dyring simulated ischemic and reperfusion.AM J Physiol 1993;264:H1897-H1906.
2. Liu Y ,Downey JM,Ischemic preconditioning protects against infarction in rat heart. Am, J physiol.1992:123:346-353.
3.Yellon DM.Alkhulaifi AM,Browne EE,et al. Ischaemic preconditioning limits infarct size in the rat heart Cardiovasc Res.1992:26:983-987
4. Zhao ZQ, Corvera JS, Halkos ME, et al..Inhibition of myocardial injury by ischemic postconditioning during reperfusion .Am J physiol.2003: 285:H579-H588
5.Kin H, Zhao ZQ, Sun HY, et al. Postconditioning attenuates myocardial ischemia-reperfusion injury by inhibiting events in the early minutes of reperfusion.Cardiovasc Res.2004:62:74-82.
6.Yang X-M,Downey JM,Cohen MV.Multiple,brief coronary occlusions during early reperfusion protect rabbit hearts by activation of ERK and production of nitric oxide. Circulation(Suppl)2003;108;IV745.
7.Downey JM, .Hsu MV, et al. Mitochondrial KATP channel opening during index ischemia and following myocardial reperfusion in ischemic rat hearts.J
Mol Cell Cardiol.2001:33:651-653.
8.Fryer RM, Hsu AK, Gross GJ. Et al.Mitochondrial KATP channel opening is important during index ischemia and following myocardial reperfusion in ischemic preconditioned rat hearts.J MOL Cell Cardiol 2001:33:831-834.
9.Grover GJ, Sleph PG, .Dzwonczyk S., et al .Role of myocardial ATP sensitive potassium channels in mediating prcconditioning in the dog heart and their possible interaction with adenosine A,-receptors.Circulation. 1992:86:1310-1316.
10.Schultz JEJ. Rose E. Yao Z .et al. Evidence for involvement of opioid receptors inchemic preconditioning in rat hearts .Am J Phvsiol.1995:268: H2157 -H2161
11.Wall TM. Sheehy R. Hartman JC, et al..Role of bradykinin in myocardial prcconditioning J pharmacol Exp ther.,1994:270:681-690.
12.Garlid, GJ,Paucek P,Yarov-yarovoy V, et al..Cardioprotective effect of diazoxide and its interaction with mitochondrial ATP-sensitive K+ channels: possible mechanisms of protection. Circ Res1997; 81:1072-1082.
13.Nakai Y, Horimito H, Mienos S, et al. Mitochondrial ATP-sensitive potassium channels play a dominant role in ischemic preconditioning of rabbit heart .Eur Surg Res.2001;33(2),57-63.
14.Hausenloy DJ, Maddock HL, Baxter GF ,et al.Inhibiting mitochondrial permeability transition pore opening : a new paradigm for myocardial preconditioning ?Cardivasc Res ,2002,55:534-543
15.Cross GJ, Auchampach JA. Block of the ATP-sensitive potassium channels prents myocardial preconditioning. Circ Res ,1992,70:223-233.,
16.Baker JE, Holman P, Gross GJ.Preconditioning in immature rabbit hearts:role of kATP channel. Circulation ,1999,99:1249-1254.
17.Rohman S, Weygandt H, Schelling P, et al..Involvement of ATP-sensitive potassium channels in preconditioning.. Basic Res cardial.1994;89:563-576.
18.Fryer RM ,Ellis JT, Hsu AK, et al. Ischemic preconditioning in rat :role mitochondrial ATP-sensitive K+ channels in the preservation of mitochondrial
fuction. Am J phsiol ,2000,278:H305-312.
19.Schultz JE, Qian YZ, Gross GJ, et al. The ischemia-selective K+ channels antagost,5-Hydroxydecanoate,.blocks ischemic preconditioning in the rat heart.J Mol Cell Cardial,1997:1050-1060.
20.Auchampach JA, Grover GJ.Blockade of ischemic preconditioning in dogs by the novel ATP dependent potassium channels angonist sodium . Cardivasc Res,1992,26:1054-1062.
21.Kita H, Miura T, Tsuchida A, et al. Suppression of reperfusion arrhythmias by preconditioning is inhibited by an ATP-sensitive potassium channels blocker 5-hygroxydecanoate but ,not by protein kinase C in the rat. J Cardiovasc phsrmacol,1998:32,791-797.
22.Crompton M,The mitochondrial permeability transition pone and its role in cell death .Biochem J, 1999,341:127-132
23.Griffiths EJ,Halestrap AP. Mitochondrial non-specific permeability transition pore opening during myocardial reperfusion
24.Crompton M.The mitochondrial permeability transition pone and its role in cell death .Biochem J, 1999,341:127-132
25 Halestrap A,connern C,Griffiths E,et al.Cyclosporin A binding to mitochondrial cyclophilin inhibits the permeability transition pore and protects hearts from ischemia/reperfusion injury. Mol Cell Biochem,1997,174:167-172
26.Halestrap AP, Clarke SJ, Javadov SA. Mitochondrial pores remain closed during ischemic but open upon reperfusion Biiochem J .1995;307:93-98
27. Kerr P, Suleiman M, Halstrap A, et al.Recorver of rat hearts reperfused after a period of ischemia is accompanied by reversal of the mitochondrial permeability transition and is enhanced by pyruvate . Am J physiol,1999, 276, H496-H502.
28.Argaud L ,Roesch OG, Raisky O ,et al.. Postconditioning inhibits mitochondrial permeability transition .Circulation 2005;111:194-197.
29..Fliss H, Gattinger D. Apoptosis in ischemic and reperfused rat myocardium. Circ Res,1996,79:949-956.
30.Zhao ZQ,Nakamura M,Wang NY, et al.Reperfusion induces myocardial apoptotic cell death.Cardiovas Res.2000;45:651-660
31.Gottlieb RA,Burlrson KO, Kloner RA,et al. Reperfusion injury induces apoptosis in rabbits cardiomyocytes. J Clin Ivest .1994;94:1621-1628. . Ann.N.Y. Acad.Sci., 1999,874:412-426.
32.Olivetti LG., Quaini F ,Sala R ,et al. Acute myocardial infarction in humans is associated with activation of programmed myocyte cell death in the surviving portion of the heart. J.Mol,Cell Cardiol.,28,2005-2016.
33.Saraste A.,Pulkki K, Kallajoki M, et al. Apoptosis in human acute myocatdial infarction.Circulation, 1997,95: 320-323
34.Gao F,.Gong B,Christopher TA, et al.Anti-apoptotic effect of benidipine,a long-lasting vasodilating calicium antagonist ,in ischemic/reperfused myocardial cells.Brit J pharm,2001,132:869-878
35 Piot CA, Padmanaban D,Ursell PC,et al Ischemic preconditioning decreases apoptosis in rat hearts in vivo Circulation,1997,96(6):1598.
36.Ichinose M, Yonemochi H, Sato T, et al.diazoxide triggers cardioprotection against apoptosis induced by oxidative stress. Am J Phsiol Heart Circ phsiol 2003,284:H2235-H2241.
37..Yang J ,Cortopassi G.. Induction of the mitochondrial permeability transition causes releaes of the apoptogenic factor cytochrome C .Free Radic Biol Med,1998,24:624-631.
38 Martin Crompton Mitochondrial intermembrane junctional complexes and their role in cell death J physiol .2000,529:11-21
39.Holmuhamedov EL, Ozcan C, Jahangir A, et al. Restoration of Ca2+ inhibited oxidative phosphorylation in cardiac mitochondria by mitochondrial Ca2+ unload. Mol cell biochem ,2001,220:135-140.
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