Landiolol, esmolol and propranolol protect from ischemia/reperfusion injury in isolated guinea pig hearts

PURPOSE: Beta blockers are thought to exert beneficial effects on the ischemic heart. The authors examined the effects of landiolol (ONO 1101), a highly selective beta1 antagonist, propranolol, a nonspecific beta blocker, and esmolol, a selective beta1 antagonist, on postischemic contractile recovery. Drugs were given prophylactically. METHODS: Ischemia-reperfusion in isolated guinea pig hearts was induced by stopping the perfusion for 45 min and reperfusing for 60 min. Hearts (n = 7 in each group) were treated with or without propranolol (1 or 10 microM), esmolol (5 or 50 microM), or landiolol (20, 100 or 500 microM) ten minutes before inducing ischemia. RESULTS: At the end of reperfusion, left ventricular pressure (LVP) recovered to 64 +/- 3% of the baseline value in the control group. With 1 and 10 microM propranolol, LVP recovered to 90 +/- 5% and 100 +/- 6% of the baseline value at 60 min after reperfusion, respectively. Fifty microM but not 5 microM of esmolol resulted in restoration of LVP to 97 +/- 17% of the pre-ischemic value at 60 min after reperfusion. In hearts pretreated with 100 and 500 microM landiolol, LVP was restored to 109 +/- 5% and 104 +/- 5% of the baseline value, respectively. Landiolol 100 microM did not depress LVP in the pre-ischemic period. CONCLUSIONS: The present study shows that landiolol, an ultra-short-acting cardioselective beta1 blocker, has cardioprotective effects on ischemia-reperfusion injury in isolated guinea pig hearts. All three beta blockers were equally protective but the intermediate dosage of landiolol preserved LVP during the pre-ischemic period.     

Cardioprotective effects of propofol in isolated ischemia-reperfused guinea pig hearts: role of KATP channels and GSK-3β

PURPOSE: Propofol exerts cardioprotective effects, but the involved mechanisms remain obscure. The present study examines the cardioprotective effects of propofol and its role in cardiac function, including its effect on K(ATP) channel opening and the inhibition of GSK-3beta activity in ischemia-reperfused hearts. METHODS: Ischemia-reperfusion (I/R) was produced in isolated guinea pig hearts by stopping coronary perfusion for 25 min, followed by reperfusion. The hearts were incubated for ten minutes, with or without propofol (25 or 50 microM), or for five minutes with 500 microM 5-hydroxydecanoate (a mitochondrial K(ATP) channel blocker) or 30 microM HMR1098 (sarcolemmal K(ATP) channel blocker), followed by five minutes with 50 microM propofol before ischemia. Action potentials on the anterior epicardial surface of the ventricle were monitored using a high-resolution charge-coupled device camera system, and at five minutes after reperfusion, GSK-3beta phosphorylation at the serine residue, Ser9, was examined. RESULTS: After 35 min of reperfusion, propofol (25 and 50 microM) blunted the adverse effects of I/R and reduced infarct size (P < 0.05). In addition, prior incubation with 5-hydroxydecanoate or HMR1098 had no effect on functional recovery improved by 50 microM propofol. At five minutes after reperfusion, propofol (25 and 50 microM) shortened the duration of the action potential and increased the levels of phospho-GSK-3beta (P < 0.05). CONCLUSIONS: Propofol enhanced mechanical cardiac recovery and reduced infarct size. The data further suggest that GSK-3beta play an important role in propofol cardioprotective actions during coronary reperfusion, but mitochondrial K(ATP) channels do not.     

Ischemic postconditioning: brief ischemia during reperfusion converts persistent ventricular fibrillation into regular rhythm

Objectives: Brief episodes of myocardial ischemia-reperfusion employed during reperfusion after a prolonged ischemic insult may attenuate the total ischemia-reperfusion injury. This phenomenon has been termed ischemic postconditioning. In the present study, we studied the possible effect of postconditioning on persistent reperfusion-induced ventricular fibrillation (VF) in the isolated rat heart model. Methods: Isolated Langendorff-perfused rat hearts (n=46) were subjected to 30 min of regional ischemia and reperfusion. The hearts with persistent VF (n=11) present after 15 min of reperfusion were then randomly assigned into one of the two groups: (1) control hearts (n=6), in which perfusion was continued without intervention; (2) postconditioned hearts (n=5) subjected to 2 min of global ischemia followed by reperfusion. Left ventricular pressures, heart rate, coronary flow, and electrogram were monitored throughout the experiment. Results: Conversion of VF into regular rhythm was observed in all hearts subjected to postconditioning. Regular beating was maintained by all postconditioned hearts during the subsequent reperfusion. None of the hearts in the control group had normal rhythm at the end of the experiment. At the end of reperfusion, the left ventricular developed pressure was lower in beating postconditioned hearts compared to the hearts that did not develop persistent VF. Conclusions: Ischemic postconditioning possesses strong antiarrhythmic effect against persistent reperfusion-induced tachyarrhythmias. Postconditioning may be an interesting, novel adjunct strategy to protect the heart.     

Effects of tramadol on myocardial ischemia-reperfusion injury

OBJECTIVES: The aim of the present study was to evaluate the effect of tramadol on isolated rat hearts subjected to global ischemia-reperfusion injury. DESIGN: Langerdorff perfused isolated rat hearts were subjected to 60 min of global ischemia following 60 min of reperfusion. In group I and III hearts were received tramadol before the onset of ischemia. Following the ischemic period, group II and III hearts were received tramadol infusion. Group I and IV hearts were subjected to saline at the same time point. The myocardial postischemic recovery was compared using hemodynamic, coronary flow, biochemical parameters from coronary effluent, and oxidative stress markers from heart tissue homogenates. RESULTS: There were significant differences between tramadol and saline used groups in hemodynamic parameters. GPx values of groups I and III were significantly lower than group IV (p<0.05). SOD values of groups I, II and III were higher than group IV (p<0.05). LDH values of groups I and II were significantly lower than groups III and IV (p<0.05). CONCLUSION: Tramadol provides a cardioprotective effect against myocardial ischemia-reperfusion in isolated rat heart.     

Anesthetic preconditioning enhances Ca2+ handling and mechanical and metabolic function elicited by Na+-Ca2+ exchange inhibition in isolated hearts

BACKGROUND: Anesthetic preconditioning (APC) is well known to protect against myocardial ischemia-reperfusion injury. Studies also show the benefit of Na+-Ca2+ exchange inhibition on ischemia-reperfusion injury. The authors tested whether APC plus Na+-Ca2+ exchange inhibitors given just on reperfusion affords additive protection in intact hearts. METHODS: Cytosolic [Ca2+] was measured by fluorescence at the left ventricular wall of guinea pig isolated hearts using indo-1 dye. Sarcoplasmic reticular Ca2+-cycling proteins, i.e., Ca2+ release channel (ryanodine receptor [RyR2]), sarcoplasmic reticular Ca2+-pump adenosine triphosphatase (SERCA2a), and phospholamban were measured by Western blots. Hearts were assigned to seven groups (n = 8 each): (1) time control; (2) ischemia; (3, 4) 10 microM Na+-Ca2+ exchange inhibitor KB-R7943 (KBR) or 1 microM SEA0400 (SEA), given during the first 10 min of reperfusion; (5) APC initiated by sevoflurane (2.2%, 0.41 +/- 0.03 mm) given for 15 min and washed out for 15 min before ischemia-reperfusion; (6, 7) APC plus KBR or SEA. RESULTS: The authors found that APC reduced the increase in systolic [Ca2+], whereas KBR and SEA both reduced the increase in diastolic [Ca2+] on reperfusion. Each intervention improved recovery of left ventricular function. Moreover, APC plus KBR or SEA afforded better functional recovery than APC, KBR, or SEA alone (P < 0.05). Ischemia-reperfusion-induced degradation of major sarcoplasmic reticular Ca2+-cycling proteins was attenuated by APC, but not by KBR or SEA. CONCLUSIONS: APC plus Na+-Ca2+ exchange inhibition exerts additive protection in part by reducing systolic and diastolic Ca2+ overload, respectively, during ischemia-reperfusion. Less degradation of sarcoplasmic reticular Ca2+-cycling proteins may also contribute to cardiac protection.     

Myocardial protection by coronary washout during global ischemic cardiac arrest

The effect of intermittent coronary washout (WO) during global ischemic cardiac arrest (ICA) was evaluated in isolated blood perfused dog hearts undergoing 90 min normothermic ICA and 90 min reperfusion. WO consisted of infusion of 100 ml normothermic dog plasma at 100 mm Hg every 10 min during ICA. Systolic and diastolic pressures were measured at constant volume with a left ventricular balloon. Coronary blood flow (CBF) was measured, and transmyocardial oxygen, lactate, and glucose differences were calculated. Adenosine triphosphate (ATP), creatine phosphate (CP), calcium (Ca²⁺), glycogen, and water content were measured from left ventricular biopsies. During 90 min of WO, hearts extracted glucose (15 ± 4 mg/g dry wt LV) and lost lactate (90 ± 5 μmole/g dry wt LV). ATP and CP were significantly depressed at arrest in both groups although WO resulted in significantly higher levels of ATP and CP. CP returned to control levels at 90 min reperfusion in the WO group but remained depressed in the ICA group. Calcium accumulation was greater in hearts with ICA. With reperfusion, both ICA and WO groups demonstrated early hyperemia and rapid lactate washout. There were no differences in total CBF, AVO₂, MOV₂, and myocardial glycogen or water content. Systolic performance was equally depressed in both groups at 90 min of reperfusion. Diastolic compliance, while still impaired, was better preserved in WO when measured in the arrested heart at 90 min ICA (P < 0.01) and in the contracting heart after 90 min reperfusion (P < 0.02). Normothermic coronary washout during 90 min of normothermic ischemic arrest without cardioplegic agents produces improved ventricular compliance and energy metabolism, and reduces myocardial calcium accumulation.     

酸处理对大鼠离体心脏缺血再灌注损伤的影响及PI3K/Akt信号通路在其中的作用

目的 评价酸处理对大鼠离体心脏缺血再灌注损伤的影响及磷脂酰肌醇-3激酶/蛋白质丝氨酸苏氨酸激酶(PI3K/Akt)信号通路在其中的作用.方法 清洁级SD大鼠70只,雌雄不拘,体重450-550 g,随机分为7组(n=10):缺血再灌注组(I/R组)、缺血后处理组(IPO组)、酸处理组(H+组)、缺血后处理+碱处理组(IPO+OH-组)、碱处理组(OH-组)、酸处理+渥曼青霉索组(H++wort组)和渥曼青霉素组(wort组).采用Langendorff装置行离体心脏灌注,采用全心停灌30 min、再灌注中性K-H液(pH值7.4)120 min的方法 制备大鼠离体心脏缺血再灌注模型.IPO组于再灌注即刻灌注中性K-H液15 s,停灌15 s,重复6次,行缺血后处理;H+组于再灌注即刻灌注经80%O2-20%CO2饱和的酸性K-H液(pH值6.9)3 min;IPO+OH-组于再灌注即刻灌注经100%O2饱和的碱性K-H液(pH值7.8)3min,并行缺血后处理;OH-组于再灌注即刻灌注碱性K-H液3 min;H++wort组于再灌注即刻灌注含100 nmol/L渥曼青霉素的酸性K-H液3 min;wort组于再灌注即刻灌注含100 nmol/L渥曼青霉素的中性K-H液3 min.于缺血前和再灌注30 min时记录左心室舒张末期压(LVEDP)和左心室压力最大上升和下降速率(±dp/dtmax);于再灌注30 min时测定冠状动脉流出液一氧化氮(NO)浓度;于再灌注120 min时,计算心肌梗塞区与缺血危险区的质量比,来表示心肌梗塞范围.结果 与I/R组比较,IPO组和H+组再灌注时LVEDP降低,±dp/dtmax升高,心肌梗塞范围减小,冠状动脉流出液NO浓度升高,OH-组、H++wort组心肌梗塞范围增加,wort组心肌梗塞范围增加,冠状动脉流出液NO浓度降低,IPO+OH-组LVEDP降低(P<0.05或0.01),±dp/dtmax、心肌梗塞范围和冠状动脉流出液NO浓度差异无统计学意义(P>0.05);IPO组与H+组上述指标比较差异均无统计学意义(P>0.05).结论 酸处理可减轻大鼠离体心脏缺血再灌注损伤.其机制与激活PI3K/Akt信号通路有关.     

Effects of tramadol on myocardial ischemia-reperfusion injury

Objectives. The aim of the present study was to evaluate the effect of tramadol on isolated rat hearts subjected to global ischemia-reperfusion injury. Design. Langerdorff perfused isolated rat hearts were subjected to 60 min of global ischemia following 60 min of reperfusion. In group I and III hearts were received tramadol before the onset of ischemia. Following the ischemic period, group II and III hearts were received tramadol infusion. Group I and IV hearts were subjected to saline at the same time point. The myocardial postischemic recovery was compared using hemodynamic, coronary flow, biochemical parameters from coronary effluent, and oxidative stress markers from heart tissue homogenates. Results. There were significant differences between tramadol and saline used groups in hemodynamic parameters. GPx values of groups I and III were significantly lower than group IV (p<0.05). SOD values of groups I, II and III were higher than group IV (p<0.05). LDH values of groups I and II were significantly lower than groups III and IV (p<0.05). Conclusion. Tramadol provides a cardioprotective effect against myocardial ischemia-reperfusion in isolated rat heart.     

Effects of inhibition of myocardial extracellular-responsive kinase and P38 mitogen-activated protein kinase on mechanical function of rat hearts after prolonged hypothermic ischemia

BACKGROUND: Mitogen-activated protein kinases (MAPKs), including extracellular-responsive kinase (ERK) and p38 MAPK, are activated by stresses associated with hypothermia-rewarming and ischemia-reperfusion. Their activation in heart is associated with beneficial (preconditioning) and adverse effects (apoptosis and impaired contractility). This study determined whether ERK and p38 MAPK activities are altered by hypothermic ischemia and normothermic reperfusion and the consequences of their inhibition on recovery of myocardial function. METHODS: Left ventricular work (L x min(-1) x mm Hg) was assessed during normothermic perfusion (30 min) of isolated rat hearts that were either freshly excised or previously subjected to hypothermic storage (8 hr, 3 degrees C) and rewarming (10 min, 37 degrees C) before normothermic reperfusion (30 min). Phospho-specific immunoblot analysis of p38 MAPK was performed in hearts and various cultured cells. RESULTS: Compared with fresh hearts, hearts subjected to hypothermia and rewarming demonstrated impaired left ventricular work (1.96+/-0.53, n=12 vs. 8.37+/-0.46, n=4, <0.05) during reperfusion. The ERK inhibitor, PD98059 (20 microM), present during storage and rewarming, caused modest improvement (3.66+/-0.75, n=9, <0.05). The p38 MAPK inhibitor, SB202190 (10 microM), when present during reperfusion, improved recovery (to 6.12+/-0.75, n=6, <0.05); it was ineffective if present only during rewarming (1.52+/-0.88, n=4). In rat2 fibroblasts, hypothermia and rewarming activated p38 MAPK and its downstream kinase MAPK-activated protein kinase 2, but not c-Jun N-terminal kinase/stress-activated protein kinase. CONCLUSIONS: Myocardial p38 MAPK and MAPK-activated protein kinase 2 are stimulated by hypothermia, ischemia, and rewarming and are detrimental to recovery of mechanical function of hearts subjected to prolonged hypothermic storage. Inhibition of p38 MAPK may be useful in protocols to improve the recovery of mechanical function of cold-stored hearts.     

右美托咪啶预处理对大鼠离体心脏缺血再灌注损伤的影响

目的 评价右美托咪啶预处理对大鼠离体心脏缺血再灌注损伤的影响.方法 健康清洁级雄性Wistar大鼠24只,体重230～ 260 g,制备离体Langendorff心脏灌注模型后,采用随机数字表法,将离体心脏随机分为3组(n=8):缺血再灌注组(I/R组)、右美托咪啶Ⅰ组(DI组)、右美托咪啶Ⅱ组(DⅡ组).各组均先用K-H液平衡灌注10 min后,I/R组用K-H液继续灌注30 min,D I组和DⅡ组分别用含有0.23.、2.30ng/ml右美托咪啶的K-H液继续灌注20 min,再用K-H液冲洗10 min.各组心脏均缺血30 min,K-H液再灌注120 min.于平衡灌注末、再灌注5、30、60和120min时收集冠脉流出液,测定肌酸激酶(CK)和乳酸脱氢酶(LDH)活性.再灌注末取心肌组织,测定SOD活性及MDA含量.结果 与I/R组比较DⅠ组和DⅡ组冠脉流出液CK、LDH活性、心肌组织MDA含量降低,心肌组织SOD活性升高(P＜0.05);与DI组比较,DⅡ组冠脉流出液CK、LDH活性、心肌组织MDA含量降低,心肌组织SOD活性升高(P＜0.05).结论 右美托咪啶预处理可减轻大鼠心肌缺血再灌注损伤,且与浓度有关.     

缺血-再灌注不同时期应用1,6-二磷酸果糖对幼兔心脏的保护作用

目的　研究在缺血 再灌注的不同时期 1,6 二磷酸果糖 (FDP)对幼兔离体心脏的保护作用。方法　建立幼兔离体心脏做功模型 ,行缺血 再灌注试验 ,于缺血 再灌注的不同时期给予5mMFDP ,观察再灌注期的心功能、心肌组织含水量、心肌MDA、冠脉液CK含量以及心肌超微结构的变化。结果　缺血期给药组与缺血前给药组和再灌注期给药组比较 ,缺血期给药组的心功能恢复百分率、心肌超微结构优于后两组 ,缺血期给药组的冠脉液CK含量、心肌组织含水量和MDA含量显著低于后两组。结论　缺血期给 1,6 二磷酸果糖对幼兔心肌缺血 再灌注损伤的保护优于缺血前或者再灌注期给药     

七氟醚后处理对大鼠离体心脏缺血再灌注时心肌细胞凋亡的影响

目的 探讨七氟醚后处理对大鼠离体心脏缺血再灌注(IR)时心肌细胞凋亡的影响.方法 雄性SD大鼠48只,随机分4组(n=12),制备离体心脏灌注模型,K-H液平衡灌注30 min后,C组灌注K-H液120 min;IR组缺血30 min,K-H液再灌注90 min;缺血后处理组(IP组)缺血30 min后行4个循环复灌20 s/缺血20 s,再灌注K-H液;七氟醚后处理组(SP组)缺血30 min后灌注含七氟醚的K-H液5 min,K-H液冲洗10 min,再灌注K-H液.IP组和SP组再灌注总时间90 min.灌注期间测定心功能,灌注结束时取心脏测定心梗面积、Bcl-2、细胞色素C和Caspase-3蛋白表达水平,计算心肌细胞凋亡指数(AI).结果 与C组比较,其余各组左心室最大上升或下降速率(±dp/dt)、左心室发展压(LVDP)、HR降低,左心室舒张末压(LVEDP)和AI升高,Bel-2、细胞色素C和Caspase-3蛋白表达上调(P<0.05);与IR组比较,IP组和SP组±dp/dt、LVDP升高,LVEDP和AI降低,心梗面积减小,Bcl-2表达上调,细胞色素C和Caspase-3蛋白表达下调(P<0.05).结论 七氟醚后处理可减少大鼠离体心脏IR时心肌细胞凋亡,其机制与其下调细胞色素C和Caspase-3蛋白表达,上调Bcl-2表达有关.     

PI3K-Akt信号通路在七氟醚预处理减轻大鼠离体心脏缺血再灌注损伤中的作用

目的 评价磷脂酰肌醇-3-激酶-丝氨酸/苏氨酸激酶(PI3K-Akt)信号通路在七氟醚预处理减轻大鼠离体心脏缺血再灌注损伤中的作用.方法 健康成年雄性SD大鼠96只,体重220～280g,采用随机数字表法,将其随机分为6组(n=16):假手术组(S组)、缺血再灌注组(I/R组)、七氟醚预处理组(SP组)、渥曼青霉素组(W组)、二甲基亚砜组(D组)和七氟醚预处理+渥曼青霉素组(SW组).采用Langendorff装置建立大鼠离体心脏缺血再灌注模型.S组继续灌注180 min;I/R组平衡灌注30 min,缺血30 min,恢复灌注120 min;其余各组先平衡灌注15 min,SP组、W组、DMSO组和SW组分别用含2.4%七氟醚、100 nmol/L渥曼青霉察、20 μmol/L二甲基亚砜、2.4%七氟醚和100 nmol/L渥曼青霉素的K-H液灌注10 min,然后洗脱5 min,缺血30 min,恢复灌注120 min.各组随机取8个心脏,于平衡灌注末和再灌注15 min时,记录HR、左室舒张末压(LVEDP)、左室发展压(LVDP)、左心室内压最大上升速率(+dp/dtmax)和左心室内压最大下降速率(-dp/dtmax).再灌注15 min时取心肌组织,采用TUNEL法检测细胞凋亡,计算凋亡指数;采用Western blot法测定磷酸化Akt(p-Akt)表达.再灌注120 min时,取8个心脏,采用TIC染色法测定心肌梗死体积.结果 与S组比较,其余各组HR、LVDP和±dp/dtmax降低,LVEDP升高,I/R组、SP组和D组心肌p-Akt表达上调(P＜0.05);与I/R组比较,SP组LVDP和±dp/dtmax升高,LVEDP和凋亡指数降低,心肌p-Akt表达上调,心肌梗死体积减小(P＜0.05),SW组上述指标差异无统计学意义(P＞0.05).结论 七氟醚预处理可通过激活PI3K-Akt信号通路减轻大鼠离体心脏缺血再灌注损伤.

Abstract：

Objective To investigate the role of phosphatidyl-inositol 3-kinase-Akt (PI3k-Akt) signal pathway in the attenuation of ischemia-reperfusion (I/R) injury by sevoflurane preconditioning in isolated rat hearts. Methods Ninety-six adult male SD rats weighing 220-280 g were randomly divided into 6 groups ( n = 16 each): sham operation group (group S); I/R group; sevoflurane preconditioning group (group SP); wortmannin group (group W); dimethyl sulfoxide (DMSO) group (group D) and sevoflurane preconditioning + wortmannin group (group SW) . Their hearts were excised and perfused in a Langendorff apparatus with K-H solution saturated with 95%O2-5%C02 at 37 ℃ . The hearts were continuously perfused for 180 min in group S. After 15 min of equilibration, the isolated hearts were subjected to 30 min of ischemia followed by 120 min of reperfusion in SP, W, D and SW groups. Croups SP, W, D and SW received 10 min of perfusion with K-H solution containing 2. 4% sevoflurane, 100 nmol/L wortmannin, 20 μmol/L DMSO, and 2.4% sevoflurane + 100 nmol/L wortmannin, respectively, followed by 5 min washout before I/R. Eight hearts in each group were selected and HR, left ventricular end-diabetic pressure (LVEDP), left ventricular developed pressure (LVDP), and ± dp/dtmax were recorded at the end of equilibration and at 15 min of reperfusion, Myocardial tissues were obtained at 15 min of reperfusion for determination of apoptosis (by TUNEL) and phosphorylated Akt (p-Akt) expression (by Western blot) . Another 8 hearts were selected at 120 min of reperfusion for determination of myocardial infarct size by TTC staining. Result Compared with group S, LVDP and ± dp/dt,^ were significantly decreased and LVEDP was significantly increased in groups I/R, SP, W, D and SW, and myocardial p-Akt expression was up-regulated in groups I/R, SP and D ( P ＜ 0.05). Compared with group I/R, LVDP and ± dp/dtmax were significantly increased, LVEDP and apoptosis index were significantly decreased, myocardial p-Akt expression was up-regulated, and myocardial infarct size was significantly reduced in group SP (P ＜0.05) . Conclusion Activation of PI3K-Akt signal pathway is involved in the attenuation of I/R injury by sevoflurane reconditioning in isolated rat hearts.     

Cardioprotective effects of tetrahydrobiopterin in cold heart preservation after cardiac arrest.

BACKGROUND: It has recently been shown that tetrahydrobiopterin (BH4), an essential cofactor of nitric oxide synthase (NOS), reduces ischemia-reperfusion myocardial injury. The aim of this study was to determine if supplementation with BH4 after cardiac arrest followed by cold heart preservation would exert a cardioprotective effect against ischemia-reperfusion injury. MATERIALS AND METHODS: Isolated perfused rat hearts were subjected to 4 degrees C cold ischemia and reperfusion. Hearts were treated with cold cardioplegic solution with or without BH4 just before ischemia and during the first 5 min of reperfusion period. Effects of BH4 on left ventricular function, myocardial contents of high-energy phosphates, and nitrite plus nitrate were measured in the perfusate, before ischemia and after reperfusion. Moreover, the effect of BH4 on the cold-heart preservation followed by normothermic (37 degrees C) ischemia was determined. RESULTS: BH4 improved the contractile and metabolic abnormalities in reperfused cold preserved hearts that were subjected to normothermic ischemia. Furthermore, BH4 significantly alleviated ischemic contracture during ischemia, and restored the diminished perfusate levels of nitrite plus nitrate after reperfusion. CONCLUSION: These results demonstrated that BH4 reduces ischemia-reperfusion injury in cold heart preservation. The cardioprotective effect of BH4 implies that BH4 could be a novel and effective therapeutic option in the preservation treatment of donor heart after cardiac arrest.     

Cardioprotection with resveratrol pretreatment: improved beneficial effects over standard treatment in rat hearts after global ischemia

OBJECTIVE: The major objective of the present study is to evaluate the potential role of resveratrol (RVT), a natural antioxidant found in grapes and red wine, in protecting the myocardium from the deleterious effects of ischemia-reperfusion (I/R) injury using isolated rat hearts. METHODS: Langendorff perfused isolated rat hearts were subjected to 60 min of global ischemia following 60 min of reperfusion. RVT was given according to chronic pretreatment and/or acute treatment protocols. Animals received RVT at the dose of 20 mg/kg via an intragastric tube for 14 days before the experiment and/or at the infusion concentration of 10 microM for 30 min before the onset of ischemia. The myocardial postischemic recovery was compared using hemodynamic data (peak systolic pressure, end diastolic pressure, and +dP/dtmax), coronary flow, biochemical parameters (LDH, CK-MB, cTnI, myoglobin) from coronary effluent, and oxidative stress markers (MDA, GSH, carbonyl) from heart tissue homogenates in each group. RESULTS: RVT pretreatment and treatment protocols have provided increased preservation in myocardial recovery following global ischemia compared to a non-treated group. Furthermore, the ischemic damage of myocardium was significantly lower in chronic pretreated rats than in the acutely treated group. In contrast, no significant difference was observed in cardioprotective effects of RVT between the only pretreated group, and both the pretreated and treated group throughout reperfusion. CONCLUSION: The findings from this study indicate that RVT has potent cardioprotective properties against I/R injury in rat hearts. The study also highlighted that the administration of RVT, as pretreatment, has amplified the beneficial effects over the standard treatment.     

Deletion of endothelial nitric oxide synthase exacerbates myocardial stunning in an isolated mouse heart model

BACKGROUND: While endothelial nitric oxide synthase (eNOS) is an important regulator of vascular tone, it is also constitutively expressed in cardiac myocytes and contributes to the regulation of myocardial function. The role of eNOS in ischemia-reperfusion is uncertain, however, with some studies showing beneficial effects while other studies demonstrate increased cardiac injury. We hypothesized that the beneficial effects of eNOS would predominate, and thus that targeted deletion of eNOS would exacerbate myocardial dysfunction following ischemia-reperfusion. MATERIALS AND METHODS: ENOS knockout and wild-type mouse hearts were Langendorff-perfused using Krebs bicarbonate buffer and subjected to 20 min of global normothermic ischemia followed by 30 min of reperfusion. Myocardial function was measured using a ventricular balloon to determine time to onset of contracture, left ventricular developed pressure (LVDP), left ventricular end-diastolic pressure (LVEDP), and rate-pressure product (RPP). RESUKTS: Heart rate and coronary resistance were similar in both groups during baseline and reperfusion periods. Diastolic function as determined by peak LVEDP during ischemia and final LVEDP after reperfusion were worse in the eNOS knockout group vs wild-type (114 and 31 mmHg vs 92 and 18 mmHg, P <.05). Although RPP (heart rate x LVDP), measured as an index of systolic function, was initially better in eNOS knockouts (24216 vs 16353), wild-type hearts recovered more function than did eNOS knockout hearts by the end of 30 min of reperfusion (30892 vs 20522, P <.05). CONCLUSIONS: These data suggest that the deletion of eNOS results in increased myocardial dysfunction following ischemia-reperfusion in an isolated heart model.     

Oral hypoglycemic sulfonylurea glimepiride preserves the myoprotective effects of ischemic preconditioning

BACKGROUND: To investigate whether the sulfonylurea glimepiride affects the myoprotective effects of ischemic preconditioning (IPC), isolated rabbit hearts were perfused with Krebs-Henseleit solution. METHODS: Eight hearts underwent IPC consisting of two cycles of 5 min global ischemia and reperfusion. Six hearts received a 5-min infusion of 10 microM glimepiride, six hearts received a 5-min infusion of 50 microM glimepiride, and seven hearts received a 5-min infusion of 10 microM glibenclamide before IPC. Seven hearts received a 5-min infusion of the selective mitochondrial K(ATP) channel opener diazoxide (50 microM). Other hearts received a 5-min infusion of 10 microM glimepiride (n = 6), 50 microM glimepiride (n = 6), or 10 microM glibenclamide (n = 7) before diazoxide. Seven hearts served as a control. All groups then were subjected to 1 h of regional ischemia, followed by 1 h of reperfusion. LV pressures, monophasic action potential duration (APD(50)), and infarct size were measured. RESULTS: Both IPC and diazoxide significantly prolonged APD(50) and preserved diastolic function at 60 min of reperfusion compared to control. In addition, both groups reduced infarct size compared to control. Glibenclamide, but not glimepiride reversed these effects. CONCLUSION: Glimepiride offers less cardiovascular effects than glibenclamide, possibly due to its lower affinity for the mitochondrial K(ATP) channels.     

Role of tetrahydrobiopterin on ischemia-reperfusion injury in isolated perfused rat hearts

AIM: It has recently been shown that nitric oxide synthase in the presence of suboptimal levels of tetrahydrobiopterin (BH(4)), an essential cofactor of nitric oxide synthase, may favor increased production of oxygen free radicals. This study was designed to define the role of BH(4) in myocardial ischemia-reperfusion injury. METHODS: Isolated perfused rat hearts were subjected to 37 degrees C ischemia and reperfusion. Hearts were received with BH(4) or vehicle for 5 min just before ischemia and during the first 5 min of the reperfusion period. The effects of BH(4) on left ventricular function, myocardial contents of lipid peroxidation and high energy phosphates, and levels of lactate dehydrogenase and nitrite plus nitrate in perfusate before ischemia and after reperfusion were estimated. Moreover, the effect of BH(4) given with 2,4-diamino-6-hydroxypyrimidine (DAHP), a selective inhibitor of BH(4) production, intraperitoneally 24 h before the experiments were estimated. RESULTS: BH(4) improved contractile and metabolic abnormalities in reperfused hearts. Furthermore, BH(4) significantly alleviated ischemic contracture during ischemia, and restored diminished perfusate levels of nitrite plus nitrate after reperfusion. On the other hand, DAHP-treatment aggravated ischemia-reperfusion induced functional and metabolic abnormalities. Administration of BH(4) improved DAHP-induced functional and metabolic abnormalities. CONCLUSION: Results demonstrated that BH(4) lessens ischemia-reperfusion injury in isolated perfused rat hearts. Conversely, deficiency of BH(4) seems to accelerate endothelial dysfunction and myocardial ischemia-reperfusion injury. Present data may be compatible with the hypothesis that nitric oxide synthase in the presence of insufficiency of BH(4) serve as the cause of oxidative injury.     

吡那地尔预处理对兔心脏缺血再灌注时心肌炎性反应的影响

目的 评价吡那地尔预处理对兔心脏缺血再灌注时心肌炎性反应的影响.方法 日本大耳白兔56只,随机分为4组:正常对照组(C组,n=8)、缺血再灌注组(I/R组,n=16)、吡那地尔预处理组(P组,n=16)和格列本脲组(G组,n=16).建立离体心脏Langendorff再灌注模型,灌注充氧K-H液,待心率平稳10 min时,C组取心肌组织,测定丙二醛(MDA)、补体C3a、细胞间粘附分子-1(ICAM-1)和细胞核NF-κBp65表达;I/R组继续灌注充氧K-H液40 min;P组灌注充氧K-H液10 min后,灌注10 μmol/L充氧吡那地尔30 min;G组灌注10 μmol/L充氧格列本脲10 min后,再灌注10 μmol/L充氧吡那地尔30min.随后I/R组、P组和G组行全心缺血40 min,再灌注充氧K-H液.I/R组、P组和G组分别于心率平稳10 min、再灌注30、60 min时取8个心脏,测定冠状静脉流出液中肌酸激酶(CK)活性和肿瘤坏死因子-α(TNF-α)浓度;再灌注60或120 min时测定心肌组织MDA、补体C3a、ICAM-1及细胞核NF-κBp65表达.结果 P组再灌注期间CK、TNF-α、MDA、补体C3a、ICAM-1和NF-κBp65水平均明显低于其他各组(P＜0.05).结论 吡那地尔预处理可减轻兔心脏缺血再灌注时心肌炎性反应,可能与ATP敏感性钾通道开放有关,从而对心肌产生保护作用.     

Na(+)/H(+) exchanger inhibitor HOE642 offers myoprotection in senescent myocardium independent of ischemic preconditioning mechanisms

BACKGROUND: Inhibition of Na(+)/H(+) exchange has been shown to provide functional protection during ischemia and reperfusion in mature heart. This study was undertaken to elucidate the effect of Na(+)/H(+) exchange inhibitor HOE642 in the aged rabbit heart. METHODS: Isolated rabbit hearts were subjected to 1 h of left descending coronary artery (LAD) ischemia and 1 h of reperfusion. To determine the effects of HOE642 on ischemia/reperfusion injury, seven aged or mature hearts received the Na(+)/H(+) exchange inhibitor HOE642 (1 microM) for 15 min before the ischemia and for 30 min after reperfusion. Seven aged (more than 135 weeks) or mature (15-20 weeks) rabbit hearts served as a control (untreated) with no interventions. Left ventricular pressures, monophasic action potentials and coronary flows were measured throughout the experiment and infarct size was detected at the end of experiment. RESULTS: (1) In the mature hearts, HOE642 improved postischemic functional recovery (63.1 +/- 5.0% vs. 84.4 +/- 5.4%, mature untreated vs. mature HOE, p < 0.05) and reduced infarct size as compared to untreated hearts (42.0 +/- 2.5% vs. 24.8 +/- 2.3%, mature untreated vs. mature HOE, p < 0.05). (2) Although infarct size in aged untreated hearts was significantly decreased as compared to mature untreated hearts (42.0 +/- 2.5% vs. 19.3 +/- 1.6%, mature untreated vs. aged untreated, p < 0.05), there are no significant differences regarding postischemic functional recovery between mature and aged untreated hearts (63.1 +/- 5.0% vs. 59.5 +/- 5.9%, mature untreated vs. aged untreated, p = n.s.). (3) In the aged hearts, HOE642 improved postischemic functional recovery as compared to untreated hearts (59.5 +/- 5.9% vs. 85.9 +/- 8.1%, aged untreated vs. aged HOE, p < 0.05). CONCLUSION: Na(+)/H(+) exchange inhibitor HOE642 is effective against ischemia-reperfusion injury in senescent as well as mature hearts.