培哚普利对大鼠心肌缺血—再灌注损伤早期保护机制研究
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摘要
研究背景：随着介入治疗的迅速发展,心肌缺血再灌注损伤成为人们研究的热点之一。目前,缺血再灌注损伤发生的机制尚未完全阐明,氧自由基生成、血管内皮损伤、钙超载和白细胞激活已较为公认,但并不能解释全部现象。最近的研究显示,心肌缺血再灌注损伤还与炎性因子产生、细胞凋亡、粘附分子表达及核转录因子调控等机制有关。有研究显示再灌注启动了一个典型的炎症反应,多形核中性粒细胞的浸润,已成为心肌缺血再灌注后的主要问题。Toll样受体4是新近发现的天然免疫中的细胞跨膜受体及病原模式识别受体,其除了参与病原微生物触发的炎症反应外,有研究提示TLR-4也参与了缺血组织损伤的炎症反应。在正常生理情况下,内皮细胞通过组成型一氧化氮合酶持续合成少量一氧化氮,对维持正常的血管内皮功能起重要作用。近年发现NO减少亦是PMN激活的一个重要因素,心肌缺血再灌注早期可导致内皮细胞功能障碍,内源性NO合成减少,抗PMN作用减弱,加重心肌缺血再灌注损伤。
目的：培哚普利在动物实验有抗心肌缺血再灌注损伤作用,减轻心肌缺血再灌注损伤引起的心肌细胞结构的改变、心肌出血、心肌顿抑以及心律失常等,但其心肌保护作用的具体机制尚待进一步研究。
本研究分两部分,第一部分观察培哚普利对缺血再灌注早期Toll样受体4蛋白的影响,探讨培哚普利对心肌缺血再灌注损伤保护作用与Toll样受体4蛋白介导的炎症反应的关系；第二部分腹腔注射NO非选择性阻断剂(L-NAME),观察培哚普利对心肌缺血再灌注损伤保护作用与NO的关系,初步探讨培哚普利对心肌缺血再灌注损伤的可能保护机制。
第一部分：
方法：30只Wistar大鼠随机分为3组：假手术组,缺血再灌注组及培哚普利组。培哚普利组灌服培哚普利4mg/kg,每日1次,共7天。假手术组和缺血再灌注组分别灌服相应体积的生理盐水。采用大鼠左冠状动脉前降支结扎法建立大鼠缺血再灌注模型,缺血30分钟,再灌注1小时。于实验结束时,取出心脏于4%多聚甲醛固定、石蜡包埋切片,HE染色观察心肌病理形态学变化,按TUNEL法检测凋亡细胞,以S-P免疫组化法检测TLR4蛋白表达。
结果：①与缺血再灌注组相比,培哚普利组炎症损伤减轻；②培哚普利组心肌细胞凋亡指数明显低于缺血再灌注组但高于假手术组(P<0.05);③培哚普利组心肌组织Toll样受体4蛋白平均积分光密度值低于缺血再灌注组但高于假手术组(P<0.05)。
第二部分：
方法：40只Wistar大鼠随机分为4组：假手术组,缺血再灌注组,培哚普利组及联合用药组。假手术组和缺血再灌注组灌服和腹腔注射等量的生理盐水,假手术组只在左冠状动脉前降支下穿线,不结扎；培哚普利组灌服培哚普利4mg/kg,腹腔注射等量的生理盐水；联合用药组灌服培哚普利4mg/kg,腹腔注射L-NAME 15mg/kg;每日1次,共7天。结扎大鼠左冠状动脉前降支,建立大鼠缺血再灌注模型。测定血流动力学参数,左室收缩压(LVSP)、左室舒张末压(LVEDP)、左室内压最大上升速率(+dP／dTmax)及左室内压最大下降速率(-dP／dTmax)。光镜观察心肌形态学改变,TUNEL法检测细胞凋亡指数。
结果：①缺血前四组动物LVSP、LVEDP、+dP/dTmax、-dP/dTmax,均无统计学差异(P>0.05)；假手术组在缺血30 min及再灌注30 min,60min时血流动力学平稳,无统计学差异(P>0.05)；缺血30 min后与缺血前相比三组动物LVSP、+dP/dTmax及-dP/dTmax均明显下降(P<0.05),但后三组间无统计学差异(P>0.05)；再灌注30 min,60min后缺血再灌注组及联合用药组LVSP+dP/dTmax及-dP／dTmax相对稳定,培哚普利组LVSP逐渐回升,与培哚普利组相比缺血再灌注组和联合用药组有统计学差异(P<0.05)。缺血30 min后三组动物LVEDP均明显上升,与缺血前相比有统计学差异(P<0.05),后三组间无统计学差异(P>0.05),再灌注30 min,60min后缺血再灌注组及联合用药组LVEDP无明显变化,培哚普利组略有下降与缺血再灌注组及联合用药组相比有统计学意义(P<0.05)(详见表2-2)。②假手术组散在心肌纤维肿胀、变性,间质轻度水肿。缺血再灌注组及联合用药组大鼠心肌纤维断裂坏死,大量炎性细胞浸润,而培哚普利组出血、坏死损伤轻微,有少量炎性细胞浸润。③与缺血再灌注组及联合用药组相比,培哚普利可减轻心肌细胞凋亡数(P<0.05),培哚普利组及联合用药组凋亡细胞数也有统计学差异(P<0.05),均明显高于假手术组(P<0.05)。
结论：
①缺血再灌注过程中存在细胞凋亡,细胞凋亡和Toll样受体4的表达是一个动态的过程。培哚普利可减少在体大鼠心肌缺血再灌注心肌细胞调亡指数,可能机制之一是通过减少Toll样受体4蛋白而减轻白细胞介素-6,TNF-α等表达,减轻炎症反应,保护缺血再灌注心肌。
②培哚普利的心肌保护作用可部分被腹腔注射NO非选择性抑制剂L-NAME所抑制,提示培哚普利对心急缺血再灌注损伤的保护作用可能与NO有关。
Background:Myocardial ischemia-reperfusion injury has been one of the study focus with the rapid development of percutaneous coronary intervention. At present, the mechanism of ischemia-reperfusion injury has not entirely beem clarified. That oxygen free radical generation, vascular endothelial injury, calcium overload and leukocyte activation may be the major mechanism, but it can't not explain all the phenomena. Recent studies have shown that myocardial ischemia-reperfusion injury is related with the inflammatory factor production, apoptosis, adhesion molecule expression and regulation of nuclear factors and other mechanisms etc. Studies have shown that reperfusion initiated a typical inflammatory response, polymorpho-nuclear neutrophil infiltration,this became the major problem in myocardial ischemia-reperfusion.Toll-like receptor 4 is a newly discovered transmembrane receptor and pathogen pattern recognition receptors in the innate immune cells, addition to participating in the inflammatory response triggered by pathogenic micro-organisms, there are research suggests that TLR-4 may also be involved in the inflammatory response of the ischemic tissue. The endothelial cell continuously synthasize a small amount of nitric oxide by constitutive nitric oxide synthase under normal physiological circumstances. This plays an important role in the maintenance of normal vascular endothelial function. There are studies showed that the reduction of NO is an important factor in PMN activation in recent years. It can lead to endothelial cell dysfunction in myocardial ischemia reperfusion, then endogenous NO synthesis reduced, weakening the role of anti-PMN and increasing the myocardial ischemia-reperfusion injury.
Objective:Perindopril has the effect of anti-myocardial ischemia-reperfusion injury in animal experiments, it can reduce myocardial ischemia-reperfusion injury caused by the changes in the structure of myocardial cells, myocardial hemorrhage, myocardial stunning, and arrhythmias, etc. The specific mechanism of myocardial protection remains to be further studied.
This study can be divided into two parts:The first study was undertaken to study the effect of Perindopril on the expression of toll like receptor-4 in rats with early myocardial ischemia-reperfusion injury and to explor the possible mechanisms of Perindopril on inhibition of myocardial ischemia-reperfusion injury. The second part was to determine the relationship between Perindopril on myocardial ischemia-reperfusion injury and the NO protective effect on myocardial ischemia-reperfusion injury by intraperitoneal injection of non-selective NO inhibitor (L-NAME), to explore the early protection mechanism of perindopril on myocardial ischemia reperfusion injury.
Part one:
Methods:Thirty Wistar rats were randomly divided into 3 group:sham operation group, myocardial IR group and perindopril group. In the perindopril group,The rats were treated with perindopril 4mg/kg,once a day,for 7 days and equal quantity of normal saline were treated in IR group and sham-operated group. The left anterior descending artery in Wister rats was ligated to establish IR model. Wistar rats were subjected to 30 minutes of coronary ligation, followed by 1 hours of reperfusion.Morphology of myocardial tissue was observed with optics microscope, the numbers of apoptotic cardiomyocyte was determined by Tunnel staining and TLR4 expressions was detected by immunohistochemistry.
Result:①Compared with IR group, Inflammatory injury in perindopril group was significantly lower than that in IR group;②Cardiomyocyte apoptosis index (AI) in perindopril group was significantly lower than that in IR group but higher than that in the sham-operated group(P< 0.05);③TLR-4 in perindopril group was significantly lower than that in the IR group but higher than that in the sham-operated group(P< 0.05).
Part Two:
Methods:40 Wistar rats were randomly divided into four groups:sham operation group, ischemic reperfusion group, perindopril group and combination group. Sham operation group and ischemic reperfusion group were drenched equal quantity of normal saline;perindopril group were drenched perindopril 4mg/kg, cobination group were drenched perindopril 4mg/kg, intraperitoneal injection of L-NAME 15mg/kg; once a day,7 days. The left anterior descending coronary artery in Wistar rats were ligated to establish ischemia-reperfusion model, the determination of hemodynamic parameters, left ventricular systolic pressure (LVSP), left ventricular end-diastolic pressure (LVEDP), left ventricular pressure maximum rate of rise and fall (+dP/dTmax and-dP/dTmax) were measured. Morphology of myocardial tissue was observed with optics microscope, the number of apoptotic cardiomyovyte were determined by Tunnel staining.
Result:(1) LVSP、LVEDP、+dP/dTmax、-dP/dTmax were no statistically significant differences among the four groups of animals before the LAD in Wistar rats were ligated (P> 0.05); There were no statistically significant differences when the LAD in Wistar rats were ligated 30 min and that were reperfused 30 min,60min(P< 0.05); There were significantly decreased among the late three groups of animals when the LAD in Wistar rats were ligated 30 min among the late three groups of animals(P< 0.05) and there were no statistically significant differences among the late three groups of animals(P>0.05); There were no statistically significant differences in ischemic reperfusion group and and combination group when the LAD in Wistar rats were reperfused 30 min,60min(P>0.05); there were increased significantly in perindopril group when the LAD in Wistar rats were reperfused 30 min,60min(P< 0.05), ischemic reperfusion group and combination group were no statistically significant differences (P>0.05)。LVEDP in the late three groups were significantly increased when the LAD in Wistar rats were ligated 30 min and there were no statistically significant differences among the three groups of animals(P> 0.05); LVEDP were no significant change in ischemic reperfusion group and combination group when the LAD in Wistar rats were reperfused 30 min,60min(P> 0.05); LVEDP decreased slightly in the perindopril group,there was statistically significant in perindopril group with ischemia-reperfusion group and the combination group and combination group (P<0.05); (2) Myocardial fibers swelling degeneration and interstitial edema were little found in sham operation group. Myocardial fibers fracture, necrosis and lots of inflammatory cells were found in the ischemia-reperfusion group and combined group, but hemorrhage and necrosis were attenuated and a little inflammatory cells infiltrating were found in perindopril group. (3) Compared with ischemic reperfusion group and combined group, perindopril can significantly reduce myocardial cell apoptosis(P< 0.05), there were statistically significant difference in perindopril group and combination group (P< 0.05), there were statistically significant difference in sham operation group and ischemic reperfusion group, perindopril group, combination group (P< 0.05).
Conclusion:
(1) Apoptosis can be found in cardiocytes during ischemia-reperfusion. Myocardial apoptosis and the expression of Toll-like receptor 4 are dynamic changing in ischemia-reperfusion injury. Perindopril could reduce cardiomyocyte apoptosis following ischemia-reperfusion, The mechan-ism may be by inhibiting the expression of inflammatory response which was induced by Toll-like receptor 4.
(2)Perindopril could reduce cardiomyocyte apoptosis following ischemia reperfusion, the myocardial protective effect of perindopril can partially be inhibited by L-NAME, a non-selective NO inhibitor.its relationship between NO and the myocardial protective effect were obvious,but the exat mechanism remains to be further studied.
目的：培哚普利在动物实验有抗心肌缺血再灌注损伤作用,减轻心肌缺血再灌注损伤引起的心肌细胞结构的改变、心肌出血、心肌顿抑以及心律失常等,但其心肌保护作用的具体机制尚待进一步研究。
本研究分两部分,第一部分观察培哚普利对缺血再灌注早期Toll样受体4蛋白的影响,探讨培哚普利对心肌缺血再灌注损伤保护作用与Toll样受体4蛋白介导的炎症反应的关系；第二部分腹腔注射NO非选择性阻断剂(L-NAME),观察培哚普利对心肌缺血再灌注损伤保护作用与NO的关系,初步探讨培哚普利对心肌缺血再灌注损伤的可能保护机制。
第一部分：
方法：30只Wistar大鼠随机分为3组：假手术组,缺血再灌注组及培哚普利组。培哚普利组灌服培哚普利4mg/kg,每日1次,共7天。假手术组和缺血再灌注组分别灌服相应体积的生理盐水。采用大鼠左冠状动脉前降支结扎法建立大鼠缺血再灌注模型,缺血30分钟,再灌注1小时。于实验结束时,取出心脏于4%多聚甲醛固定、石蜡包埋切片,HE染色观察心肌病理形态学变化,按TUNEL法检测凋亡细胞,以S-P免疫组化法检测TLR4蛋白表达。
结果：①与缺血再灌注组相比,培哚普利组炎症损伤减轻；②培哚普利组心肌细胞凋亡指数明显低于缺血再灌注组但高于假手术组(P<0.05);③培哚普利组心肌组织Toll样受体4蛋白平均积分光密度值低于缺血再灌注组但高于假手术组(P<0.05)。
第二部分：
方法：40只Wistar大鼠随机分为4组：假手术组,缺血再灌注组,培哚普利组及联合用药组。假手术组和缺血再灌注组灌服和腹腔注射等量的生理盐水,假手术组只在左冠状动脉前降支下穿线,不结扎；培哚普利组灌服培哚普利4mg/kg,腹腔注射等量的生理盐水；联合用药组灌服培哚普利4mg/kg,腹腔注射L-NAME 15mg/kg;每日1次,共7天。结扎大鼠左冠状动脉前降支,建立大鼠缺血再灌注模型。测定血流动力学参数,左室收缩压(LVSP)、左室舒张末压(LVEDP)、左室内压最大上升速率(+dP／dTmax)及左室内压最大下降速率(-dP／dTmax)。光镜观察心肌形态学改变,TUNEL法检测细胞凋亡指数。
结果：①缺血前四组动物LVSP、LVEDP、+dP/dTmax、-dP/dTmax,均无统计学差异(P>0.05)；假手术组在缺血30 min及再灌注30 min,60min时血流动力学平稳,无统计学差异(P>0.05)；缺血30 min后与缺血前相比三组动物LVSP、+dP/dTmax及-dP/dTmax均明显下降(P<0.05),但后三组间无统计学差异(P>0.05)；再灌注30 min,60min后缺血再灌注组及联合用药组LVSP+dP/dTmax及-dP／dTmax相对稳定,培哚普利组LVSP逐渐回升,与培哚普利组相比缺血再灌注组和联合用药组有统计学差异(P<0.05)。缺血30 min后三组动物LVEDP均明显上升,与缺血前相比有统计学差异(P<0.05),后三组间无统计学差异(P>0.05),再灌注30 min,60min后缺血再灌注组及联合用药组LVEDP无明显变化,培哚普利组略有下降与缺血再灌注组及联合用药组相比有统计学意义(P<0.05)(详见表2-2)。②假手术组散在心肌纤维肿胀、变性,间质轻度水肿。缺血再灌注组及联合用药组大鼠心肌纤维断裂坏死,大量炎性细胞浸润,而培哚普利组出血、坏死损伤轻微,有少量炎性细胞浸润。③与缺血再灌注组及联合用药组相比,培哚普利可减轻心肌细胞凋亡数(P<0.05),培哚普利组及联合用药组凋亡细胞数也有统计学差异(P<0.05),均明显高于假手术组(P<0.05)。
结论：
①缺血再灌注过程中存在细胞凋亡,细胞凋亡和Toll样受体4的表达是一个动态的过程。培哚普利可减少在体大鼠心肌缺血再灌注心肌细胞调亡指数,可能机制之一是通过减少Toll样受体4蛋白而减轻白细胞介素-6,TNF-α等表达,减轻炎症反应,保护缺血再灌注心肌。
②培哚普利的心肌保护作用可部分被腹腔注射NO非选择性抑制剂L-NAME所抑制,提示培哚普利对心急缺血再灌注损伤的保护作用可能与NO有关。
Background:Myocardial ischemia-reperfusion injury has been one of the study focus with the rapid development of percutaneous coronary intervention. At present, the mechanism of ischemia-reperfusion injury has not entirely beem clarified. That oxygen free radical generation, vascular endothelial injury, calcium overload and leukocyte activation may be the major mechanism, but it can't not explain all the phenomena. Recent studies have shown that myocardial ischemia-reperfusion injury is related with the inflammatory factor production, apoptosis, adhesion molecule expression and regulation of nuclear factors and other mechanisms etc. Studies have shown that reperfusion initiated a typical inflammatory response, polymorpho-nuclear neutrophil infiltration,this became the major problem in myocardial ischemia-reperfusion.Toll-like receptor 4 is a newly discovered transmembrane receptor and pathogen pattern recognition receptors in the innate immune cells, addition to participating in the inflammatory response triggered by pathogenic micro-organisms, there are research suggests that TLR-4 may also be involved in the inflammatory response of the ischemic tissue. The endothelial cell continuously synthasize a small amount of nitric oxide by constitutive nitric oxide synthase under normal physiological circumstances. This plays an important role in the maintenance of normal vascular endothelial function. There are studies showed that the reduction of NO is an important factor in PMN activation in recent years. It can lead to endothelial cell dysfunction in myocardial ischemia reperfusion, then endogenous NO synthesis reduced, weakening the role of anti-PMN and increasing the myocardial ischemia-reperfusion injury.
Objective:Perindopril has the effect of anti-myocardial ischemia-reperfusion injury in animal experiments, it can reduce myocardial ischemia-reperfusion injury caused by the changes in the structure of myocardial cells, myocardial hemorrhage, myocardial stunning, and arrhythmias, etc. The specific mechanism of myocardial protection remains to be further studied.
This study can be divided into two parts:The first study was undertaken to study the effect of Perindopril on the expression of toll like receptor-4 in rats with early myocardial ischemia-reperfusion injury and to explor the possible mechanisms of Perindopril on inhibition of myocardial ischemia-reperfusion injury. The second part was to determine the relationship between Perindopril on myocardial ischemia-reperfusion injury and the NO protective effect on myocardial ischemia-reperfusion injury by intraperitoneal injection of non-selective NO inhibitor (L-NAME), to explore the early protection mechanism of perindopril on myocardial ischemia reperfusion injury.
Part one:
Methods:Thirty Wistar rats were randomly divided into 3 group:sham operation group, myocardial IR group and perindopril group. In the perindopril group,The rats were treated with perindopril 4mg/kg,once a day,for 7 days and equal quantity of normal saline were treated in IR group and sham-operated group. The left anterior descending artery in Wister rats was ligated to establish IR model. Wistar rats were subjected to 30 minutes of coronary ligation, followed by 1 hours of reperfusion.Morphology of myocardial tissue was observed with optics microscope, the numbers of apoptotic cardiomyocyte was determined by Tunnel staining and TLR4 expressions was detected by immunohistochemistry.
Result:①Compared with IR group, Inflammatory injury in perindopril group was significantly lower than that in IR group;②Cardiomyocyte apoptosis index (AI) in perindopril group was significantly lower than that in IR group but higher than that in the sham-operated group(P< 0.05);③TLR-4 in perindopril group was significantly lower than that in the IR group but higher than that in the sham-operated group(P< 0.05).
Part Two:
Methods:40 Wistar rats were randomly divided into four groups:sham operation group, ischemic reperfusion group, perindopril group and combination group. Sham operation group and ischemic reperfusion group were drenched equal quantity of normal saline;perindopril group were drenched perindopril 4mg/kg, cobination group were drenched perindopril 4mg/kg, intraperitoneal injection of L-NAME 15mg/kg; once a day,7 days. The left anterior descending coronary artery in Wistar rats were ligated to establish ischemia-reperfusion model, the determination of hemodynamic parameters, left ventricular systolic pressure (LVSP), left ventricular end-diastolic pressure (LVEDP), left ventricular pressure maximum rate of rise and fall (+dP/dTmax and-dP/dTmax) were measured. Morphology of myocardial tissue was observed with optics microscope, the number of apoptotic cardiomyovyte were determined by Tunnel staining.
Result:(1) LVSP、LVEDP、+dP/dTmax、-dP/dTmax were no statistically significant differences among the four groups of animals before the LAD in Wistar rats were ligated (P> 0.05); There were no statistically significant differences when the LAD in Wistar rats were ligated 30 min and that were reperfused 30 min,60min(P< 0.05); There were significantly decreased among the late three groups of animals when the LAD in Wistar rats were ligated 30 min among the late three groups of animals(P< 0.05) and there were no statistically significant differences among the late three groups of animals(P>0.05); There were no statistically significant differences in ischemic reperfusion group and and combination group when the LAD in Wistar rats were reperfused 30 min,60min(P>0.05); there were increased significantly in perindopril group when the LAD in Wistar rats were reperfused 30 min,60min(P< 0.05), ischemic reperfusion group and combination group were no statistically significant differences (P>0.05)。LVEDP in the late three groups were significantly increased when the LAD in Wistar rats were ligated 30 min and there were no statistically significant differences among the three groups of animals(P> 0.05); LVEDP were no significant change in ischemic reperfusion group and combination group when the LAD in Wistar rats were reperfused 30 min,60min(P> 0.05); LVEDP decreased slightly in the perindopril group,there was statistically significant in perindopril group with ischemia-reperfusion group and the combination group and combination group (P<0.05); (2) Myocardial fibers swelling degeneration and interstitial edema were little found in sham operation group. Myocardial fibers fracture, necrosis and lots of inflammatory cells were found in the ischemia-reperfusion group and combined group, but hemorrhage and necrosis were attenuated and a little inflammatory cells infiltrating were found in perindopril group. (3) Compared with ischemic reperfusion group and combined group, perindopril can significantly reduce myocardial cell apoptosis(P< 0.05), there were statistically significant difference in perindopril group and combination group (P< 0.05), there were statistically significant difference in sham operation group and ischemic reperfusion group, perindopril group, combination group (P< 0.05).
Conclusion:
(1) Apoptosis can be found in cardiocytes during ischemia-reperfusion. Myocardial apoptosis and the expression of Toll-like receptor 4 are dynamic changing in ischemia-reperfusion injury. Perindopril could reduce cardiomyocyte apoptosis following ischemia-reperfusion, The mechan-ism may be by inhibiting the expression of inflammatory response which was induced by Toll-like receptor 4.
(2)Perindopril could reduce cardiomyocyte apoptosis following ischemia reperfusion, the myocardial protective effect of perindopril can partially be inhibited by L-NAME, a non-selective NO inhibitor.its relationship between NO and the myocardial protective effect were obvious,but the exat mechanism remains to be further studied.
引文
[1]Moen SAL,Claeys MJ,Timmermans JP. Myocardial ischemia/reperfusion-injury, a clinical view on a complex pathophysiological Process.Int J Cardiol 2005,100 (2):179-190.
[2]Zhang M, Xu YJ, Saini HK, et al. Pentoxifylline attenuates cardiac dysfunction and reduces TNF-alpha level in ischemic-reperfused heart. Am J Physiol Heart Circ Physiol,2005 Aug,289(2):H832-9.
[3]Frangogiannis NQ Smith CW,Entman ML. The inflammatory response in myocardial infarction. Cardiovasc Res,2002,53:31-47.
[4]赵明中,高承梅,张宇洋,等.通心络胶囊对缺血再灌注心肌细胞凋亡及相关基因蛋白表达的影响.中华心血管病杂志,2000,28(3)：206.
[5]齐丽彤,张多均,李大云.血管肾张素型受体阻断剂抑制大鼠缺血-再灌注心肌细胞凋亡.中华心血管病杂志,2001,29(2)：118.
[6]Ray PS, Martin JL, Swanson EA, et al.Transgene overexpression of alphaB crystal-llin conefrs simultaneous portection against cardiomyoeyte apoptosis and necrosis during myocardial ischemia and reperfusion.FASEB [J],2001,15:393-402.
[7]Elsasser A, Suzuki K, Lorenz-Meyer S, et al. The The role of apoptosis in myocardial ischemia:a critical appraisal.Basic Res Cardiol2001;96(3):219-226.
[8]Palace V, Kumar D, Hill MF. et al.R egional differences in non-enzymatic antioxidants in the heart under control and oxidative stress conditions.J. Mol Cell Cardiol,1999;31(1):193-202.
[9]Scarabelli T, Stephanou A, Rayment N, et al. Apoptosis of endothelial cells Precedes myocyte cell apoptosis in ischemia/reperfusion injury. Circulation,2001;104(4):253-256.
[10]Louis WJ et al Single-dose and steady-state pharmacokinetic and pharmacodynamics of perindopril in hypertensive subjects. J Cardiovasc Pharmacol 1992.20:505
[11]Louis WJ et al Comparison of the pharmacokinetics and pharmacodynamics of perindopril,cilaxapril and enalapril. Clin exp Pharmacol,1992,19(suppl 19):55
[12]Campbell DJ et al. Bradykinin peptides in kindey,blood and other tissues of the rat. Hypertension,1993,21 (2):155
[13]马克娟,马克威,赵利华,等.Toll样受体4和肿瘤坏死因子α mRNA在动脉粥样硬化中表达的相关性研究.中华心血管病杂志,2004,32：899-902.
[14]Oyama J, Blais C Jr, Liu X, et al. Reduced myocardial ischemia-reperfusion injury intol-like receptor 4-deficient mice. Circulation.2004.109:784-789.
[15]Chong AJ, Shimamoto A, Hampton CR, et al. Tol-like receptor 4 mediates ischemia-reperfusion injury of the heart. J.Thorac Cardiovasc Surg.2004.128:170-179.
[16]杨简,杨俊,丁家望等.Toll样受体4在大鼠心肌缺血再灌注损伤中表达的实验研究.中华心血管病 杂志.2008.1(36)：57-61.
[17]Murry CE, Jeanings RB, Reimer KA. Preconditioniny with ischemia:A delay of lethal cell injury in ischemie myocardium[J].Circulation,1986,74(5):1124.
[18]Han J, Kim N, Joo H, et al. ATP-sensitive K+channel activation by nitric oxide and protein kinase G in rabbit ventricular myocytes[J]. Am J Physiol Heart CircPhysiol,2002,283(4):H1545-H1554.
[19]Matsugnchi T, Masuda A, Sugimoto K, et al. JNK-interacting protein 3 associates with Toll-like receptor 4 and is involved in LPS-mediated JNK activation. EMBOJ.2o03,22:4455-4464.
[20]Hollestelle SC, De Vries MR, Van Keulen JK,et al. Toll-like receptor 4 is involve in outward arterial remodeling. Circulation.2004 Jan 27;109(3):393-8.
[21]de Kleijn D, pasterkamp G. Toll-like recptors in cardiovascular diseases. Cardiovasc Res.2003 Oct 15;60(1):58-67.
[22]Cai H, Harrison DGEndothelial Dysfunction in Cardiovascular Diseases:The Role of Oxidant Stress. Circ Res,2000,87:840-844
[23]余跃,陈加新主编.一氧化氮与疾病.第1版.北京：科学技术出版社.1998.2-8.
[24]Ferdinandy P, Csonka C, Csont T, et al. Rapid pacing-induced preconditioning is recaptured by famesol treatment in hearts of cholestero-fed rats:role of polyprenyl derivatives and nitric oxide. Mol Cell Biochem, 1998,186:27-34.
[25]Cuongd V, Kim N, Youm JB, et al. Nitric oxide-cGMP-protein kinase G-signaling pathway induces anoxie preconditioning through activation of ATP sensitive K channels in rat hearts[J].Am J Physil Heart Circ Physiol,2006,290(5):H1808-1917.
[26]Lefer AM. Attenuation of myocardial isehemia-reperfusion injury with nitric oxide replacement therapy. AnnThorac Surg,1995,60:847-851
[27]Lefer AM, Ma XL, Weyrieh A, et al. Endothelial dysfunetion and neutrophil adherence as a critical events in the development of reperfusion injury. Agents Actions SuPPl,1993,41:127-135.
[28]Lefer DJ. Myocardial proteetive actions of nitric oxide donors after myocardial isehernia and reperfusion.New Horiz,1995,3:105-112.
[29]Jones S P.Greer J J,Kakkar A K,et al.Endothelial nitric oxide synthase overexpression attenuates myocardial reperfusion injury [J].Am JPhysiol Heart Circ Physiol,2004,286(1):H276-282.
[30]Stuart-Smith K, Demystified. Nitric oxide[J]. MolPathol,2002,55(6):360-366.
[31]Cidad P,Almeida A,Bolanos J p.Inhibition of mitochondrial respiration by nitric oxide rapidly stimulates cytoprotective GLUT3-mediated glucose uptake through 5-AMP-activated protein kinase [J]. Biochem J, 2004,384(Pt3):629-636.
[1]Yellon DM, Baxter GF. Protecting the ischemic and reperfused myocardium in acute myocardial infarction:distant dream or near reality?[J]. Heart,2000,83:1651.
[2]kark M,Ziemer G, Zoeller M et al. Protection ofthe chronic by-Poxic immature rat heart during globalisehemia. Ann Thorac Surg,1995,59:699.
[3]Frangogiannis NQSmith CW,Entman ML. The inflammatory response in myocardial infarction. Cardiovasc Res,2002,53:31-47.
[4]戴玉柱.ACEI在心血管疾病中的应用.临床内科杂志,1993,10：6
[5]叶刚,张肖明,杨方,等.心肌缺血时大鼠离体心脏局部肾素血管紧张素系统表达的变化.基础医学与临床,2001,21：43-47
[6]Kinma GT, Chun YS, Park JW, et al. Role of apoptosis-inducing factor in myocardial cell death by ischemia-reperfusion[J]. Biochem Biophys Res Commun,2003,309:619.
[7]Zhu BQ, Sievers RE, Browne AE, et al. Comparative efects of aspirin with ACE inhibitor or angiotensin receptor blocker on myocardial infarction and vascular function[J]. J Renin Angiotensin Aldosterone Syst, 2003,4(1):31.
[8]杨彦玲,师养荣,李建龙,等.心肌缺血再灌注损伤研究进展[J].心血管病学进展,2003,24(2)：116-121.
[9]Scarabelli T, Stephanou A, Rayment N,et al.Apoptosis of endothelial cell apoptosis in ischemia/rerfusion injury.Circulation,2001,104:253-256.
[10]金惠铭.病理生理学[M].第5版.北京：人民卫生出版社,2003.178.
[11]Regitz-Zagrosek V, Fielitz J, Fleck E. Myocardial angiotensln receptors in human hearts[J]. Basic Res Cardiol,1998,93(Suppl2):37-42.
[12]Wu L,lwai M, Nakagami H, et al. Efect of angiotensin Ⅱ type 1 receptor blockade on cardiac remodeling in angiotensin Ⅱ type 2 receptor null mice [J]. Arterioscler Thromb VascBiol,2002,22(1):49-54.
[13]Carey RM, WangZQ, Siragy HM. Role of the angiotensin type2 receptor in the regulation of blood pressure and renalfunction[J].-Hypertension,2000,35(1 Pt 2):155-163.
[14]Chakrabarti S,Hoque AN,Karrnazyn M.A rapid ischemia-induced apoptosis in isolated rat hearts and its attenuation by the sodium-hydrogen exchange inhibitor HOE642(cariporide) [J] J Mol Cell Cardiol,29(11):3169-3174.
[15]梁绪国潘其兴.氯沙坦与卡托普利合用对凋亡调控基因的影响[J].国外医学·心血管疾病分册,2003,30(3)：112-115.
[16]Crawford K W,Freg E A,Cote T E.Angiotensin II receptor recognized by dup regulates two distinct guanine nucleotidebinding protein signaling pathways.Mol Phamacol,1992;41:154162
[17]Ruiz JA, Lopez S, Gonzalez M, et al Reactive oxygen species induce proliferation of bovine aortic endothelial cells. J Cardiovasc Pharmacol,2000.35:109-113.
[18]An J.Varadarajan SQCamara A.et al Blocking Na+/H+exchange Reduces[Na+](i)and [Ca2+](i)load after ischemia and improves function in intact hearts.Am J Physioi Heartcirc Physiol,2001,281:2398-2409
[19]王一明 李兆志 秦志端.卡托普利对心肌缺血再灌注损伤保护作用的观察[J].中国危重病急救医学,1999.11(1)：52-53.
[20]Linger T.Gohlke P.Converting enzyme inhibitors in cardiovascular therapy:current status and future potential.Cardiovasc Res,1994;28:146-158.
[21]Jin M,Wilhelm M J,Lang R E.Endogenous tissue reninangiotensin systems from molecula biology to therapy.Am J Med,1988;84(Suppl 3A):2834.
[22]Hsuch WA,Quinones MJ.Role of Endothelial Dysfunction in insulin Resistance.Am J Cardiol,2003, 92:10J-17J
[23]Zhuo JL,Mendelsohn FA.Ohishi M.Perindol alter vascular angiotensin-converting enzyme,AT(1) receptor,and nitric oxide synthase expression in patients with coronary heart disease. Hyperten-sion,2002,39(2,pt2):634-638.
[24]Miki T, Miura T, Ura N, et al. Captopril Potentiates the Myocardial Infarct Size-limiting Effect of Ischemic Preconditioning Through Bradykinin B2 Receptor Ativation[J]. J Am Coll Cardiol,1996,28: 1616-1622.
[25]Morris S D, Yellon D M. Angiotensin-converting Enzyme Inhibitors Potentiate Preconditioning Througy Bradyikiik B2 Receptor Activation in Human Heart [J] J Am Coll Cardiol,1997,29:1599-1606.
[26]Witherow FN, Helmy A,Webb DJ,et al.Bradykinin contributes to the vasodilator effects of chronic angiotensin-converting enzyme inhibiton in patients with heart failer. Circulation,2001,104(18):2177-2181.
[27]M.C.Chappell,S.N.iyer.D.L.Diz et al. Antihypertensive effects of Angiotensin-(1-7).Braz J Med.Biol.Res,1998,31:1205-1212.
[28]Kucharewicz I,Pawlak R,Matys T.et al.Antithrombotic effect of captopril and losartan is mediated by angiotensin-(1-7).Hypertension,2002,40:774.
[29]FerreIR AJ,Santos RA,Almeida AP,Angiotension-(1-7):Cardioprotective effect in myocardial ischemia/reperfusion.Hypertension,2001,38:665:668.
[30]刘风芝,李延平黄明莉,等.冬虫夏草醇提取物对大鼠缺血再灌注过程心肌保护作用研究.中国病理生理杂志,1999;15(3)：240.
[31]郭向阳,徐守春,罗爱伦,等.卡托普利对心肌组织高能磷酸化合物代谢的影响中华胸心血管外科杂志,1998；14：115.
[2]Zhang M, Xu YJ, Saini HK, et al. Pentoxifylline attenuates cardiac dysfunction and reduces TNF-alpha level in ischemic-reperfused heart. Am J Physiol Heart Circ Physiol,2005 Aug,289(2):H832-9.
[3]Frangogiannis NQ Smith CW,Entman ML. The inflammatory response in myocardial infarction. Cardiovasc Res,2002,53:31-47.
[4]赵明中,高承梅,张宇洋,等.通心络胶囊对缺血再灌注心肌细胞凋亡及相关基因蛋白表达的影响.中华心血管病杂志,2000,28(3)：206.
[5]齐丽彤,张多均,李大云.血管肾张素型受体阻断剂抑制大鼠缺血-再灌注心肌细胞凋亡.中华心血管病杂志,2001,29(2)：118.
[6]Ray PS, Martin JL, Swanson EA, et al.Transgene overexpression of alphaB crystal-llin conefrs simultaneous portection against cardiomyoeyte apoptosis and necrosis during myocardial ischemia and reperfusion.FASEB [J],2001,15:393-402.
[7]Elsasser A, Suzuki K, Lorenz-Meyer S, et al. The The role of apoptosis in myocardial ischemia:a critical appraisal.Basic Res Cardiol2001;96(3):219-226.
[8]Palace V, Kumar D, Hill MF. et al.R egional differences in non-enzymatic antioxidants in the heart under control and oxidative stress conditions.J. Mol Cell Cardiol,1999;31(1):193-202.
[9]Scarabelli T, Stephanou A, Rayment N, et al. Apoptosis of endothelial cells Precedes myocyte cell apoptosis in ischemia/reperfusion injury. Circulation,2001;104(4):253-256.
[10]Louis WJ et al Single-dose and steady-state pharmacokinetic and pharmacodynamics of perindopril in hypertensive subjects. J Cardiovasc Pharmacol 1992.20:505
[11]Louis WJ et al Comparison of the pharmacokinetics and pharmacodynamics of perindopril,cilaxapril and enalapril. Clin exp Pharmacol,1992,19(suppl 19):55
[12]Campbell DJ et al. Bradykinin peptides in kindey,blood and other tissues of the rat. Hypertension,1993,21 (2):155
[13]马克娟,马克威,赵利华,等.Toll样受体4和肿瘤坏死因子α mRNA在动脉粥样硬化中表达的相关性研究.中华心血管病杂志,2004,32：899-902.
[14]Oyama J, Blais C Jr, Liu X, et al. Reduced myocardial ischemia-reperfusion injury intol-like receptor 4-deficient mice. Circulation.2004.109:784-789.
[15]Chong AJ, Shimamoto A, Hampton CR, et al. Tol-like receptor 4 mediates ischemia-reperfusion injury of the heart. J.Thorac Cardiovasc Surg.2004.128:170-179.
[16]杨简,杨俊,丁家望等.Toll样受体4在大鼠心肌缺血再灌注损伤中表达的实验研究.中华心血管病 杂志.2008.1(36)：57-61.
[17]Murry CE, Jeanings RB, Reimer KA. Preconditioniny with ischemia:A delay of lethal cell injury in ischemie myocardium[J].Circulation,1986,74(5):1124.
[18]Han J, Kim N, Joo H, et al. ATP-sensitive K+channel activation by nitric oxide and protein kinase G in rabbit ventricular myocytes[J]. Am J Physiol Heart CircPhysiol,2002,283(4):H1545-H1554.
[19]Matsugnchi T, Masuda A, Sugimoto K, et al. JNK-interacting protein 3 associates with Toll-like receptor 4 and is involved in LPS-mediated JNK activation. EMBOJ.2o03,22:4455-4464.
[20]Hollestelle SC, De Vries MR, Van Keulen JK,et al. Toll-like receptor 4 is involve in outward arterial remodeling. Circulation.2004 Jan 27;109(3):393-8.
[21]de Kleijn D, pasterkamp G. Toll-like recptors in cardiovascular diseases. Cardiovasc Res.2003 Oct 15;60(1):58-67.
[22]Cai H, Harrison DGEndothelial Dysfunction in Cardiovascular Diseases:The Role of Oxidant Stress. Circ Res,2000,87:840-844
[23]余跃,陈加新主编.一氧化氮与疾病.第1版.北京：科学技术出版社.1998.2-8.
[24]Ferdinandy P, Csonka C, Csont T, et al. Rapid pacing-induced preconditioning is recaptured by famesol treatment in hearts of cholestero-fed rats:role of polyprenyl derivatives and nitric oxide. Mol Cell Biochem, 1998,186:27-34.
[25]Cuongd V, Kim N, Youm JB, et al. Nitric oxide-cGMP-protein kinase G-signaling pathway induces anoxie preconditioning through activation of ATP sensitive K channels in rat hearts[J].Am J Physil Heart Circ Physiol,2006,290(5):H1808-1917.
[26]Lefer AM. Attenuation of myocardial isehemia-reperfusion injury with nitric oxide replacement therapy. AnnThorac Surg,1995,60:847-851
[27]Lefer AM, Ma XL, Weyrieh A, et al. Endothelial dysfunetion and neutrophil adherence as a critical events in the development of reperfusion injury. Agents Actions SuPPl,1993,41:127-135.
[28]Lefer DJ. Myocardial proteetive actions of nitric oxide donors after myocardial isehernia and reperfusion.New Horiz,1995,3:105-112.
[29]Jones S P.Greer J J,Kakkar A K,et al.Endothelial nitric oxide synthase overexpression attenuates myocardial reperfusion injury [J].Am JPhysiol Heart Circ Physiol,2004,286(1):H276-282.
[30]Stuart-Smith K, Demystified. Nitric oxide[J]. MolPathol,2002,55(6):360-366.
[31]Cidad P,Almeida A,Bolanos J p.Inhibition of mitochondrial respiration by nitric oxide rapidly stimulates cytoprotective GLUT3-mediated glucose uptake through 5-AMP-activated protein kinase [J]. Biochem J, 2004,384(Pt3):629-636.
[1]Yellon DM, Baxter GF. Protecting the ischemic and reperfused myocardium in acute myocardial infarction:distant dream or near reality?[J]. Heart,2000,83:1651.
[2]kark M,Ziemer G, Zoeller M et al. Protection ofthe chronic by-Poxic immature rat heart during globalisehemia. Ann Thorac Surg,1995,59:699.
[3]Frangogiannis NQSmith CW,Entman ML. The inflammatory response in myocardial infarction. Cardiovasc Res,2002,53:31-47.
[4]戴玉柱.ACEI在心血管疾病中的应用.临床内科杂志,1993,10：6
[5]叶刚,张肖明,杨方,等.心肌缺血时大鼠离体心脏局部肾素血管紧张素系统表达的变化.基础医学与临床,2001,21：43-47
[6]Kinma GT, Chun YS, Park JW, et al. Role of apoptosis-inducing factor in myocardial cell death by ischemia-reperfusion[J]. Biochem Biophys Res Commun,2003,309:619.
[7]Zhu BQ, Sievers RE, Browne AE, et al. Comparative efects of aspirin with ACE inhibitor or angiotensin receptor blocker on myocardial infarction and vascular function[J]. J Renin Angiotensin Aldosterone Syst, 2003,4(1):31.
[8]杨彦玲,师养荣,李建龙,等.心肌缺血再灌注损伤研究进展[J].心血管病学进展,2003,24(2)：116-121.
[9]Scarabelli T, Stephanou A, Rayment N,et al.Apoptosis of endothelial cell apoptosis in ischemia/rerfusion injury.Circulation,2001,104:253-256.
[10]金惠铭.病理生理学[M].第5版.北京：人民卫生出版社,2003.178.
[11]Regitz-Zagrosek V, Fielitz J, Fleck E. Myocardial angiotensln receptors in human hearts[J]. Basic Res Cardiol,1998,93(Suppl2):37-42.
[12]Wu L,lwai M, Nakagami H, et al. Efect of angiotensin Ⅱ type 1 receptor blockade on cardiac remodeling in angiotensin Ⅱ type 2 receptor null mice [J]. Arterioscler Thromb VascBiol,2002,22(1):49-54.
[13]Carey RM, WangZQ, Siragy HM. Role of the angiotensin type2 receptor in the regulation of blood pressure and renalfunction[J].-Hypertension,2000,35(1 Pt 2):155-163.
[14]Chakrabarti S,Hoque AN,Karrnazyn M.A rapid ischemia-induced apoptosis in isolated rat hearts and its attenuation by the sodium-hydrogen exchange inhibitor HOE642(cariporide) [J] J Mol Cell Cardiol,29(11):3169-3174.
[15]梁绪国潘其兴.氯沙坦与卡托普利合用对凋亡调控基因的影响[J].国外医学·心血管疾病分册,2003,30(3)：112-115.
[16]Crawford K W,Freg E A,Cote T E.Angiotensin II receptor recognized by dup regulates two distinct guanine nucleotidebinding protein signaling pathways.Mol Phamacol,1992;41:154162
[17]Ruiz JA, Lopez S, Gonzalez M, et al Reactive oxygen species induce proliferation of bovine aortic endothelial cells. J Cardiovasc Pharmacol,2000.35:109-113.
[18]An J.Varadarajan SQCamara A.et al Blocking Na+/H+exchange Reduces[Na+](i)and [Ca2+](i)load after ischemia and improves function in intact hearts.Am J Physioi Heartcirc Physiol,2001,281:2398-2409
[19]王一明 李兆志 秦志端.卡托普利对心肌缺血再灌注损伤保护作用的观察[J].中国危重病急救医学,1999.11(1)：52-53.
[20]Linger T.Gohlke P.Converting enzyme inhibitors in cardiovascular therapy:current status and future potential.Cardiovasc Res,1994;28:146-158.
[21]Jin M,Wilhelm M J,Lang R E.Endogenous tissue reninangiotensin systems from molecula biology to therapy.Am J Med,1988;84(Suppl 3A):2834.
[22]Hsuch WA,Quinones MJ.Role of Endothelial Dysfunction in insulin Resistance.Am J Cardiol,2003, 92:10J-17J
[23]Zhuo JL,Mendelsohn FA.Ohishi M.Perindol alter vascular angiotensin-converting enzyme,AT(1) receptor,and nitric oxide synthase expression in patients with coronary heart disease. Hyperten-sion,2002,39(2,pt2):634-638.
[24]Miki T, Miura T, Ura N, et al. Captopril Potentiates the Myocardial Infarct Size-limiting Effect of Ischemic Preconditioning Through Bradykinin B2 Receptor Ativation[J]. J Am Coll Cardiol,1996,28: 1616-1622.
[25]Morris S D, Yellon D M. Angiotensin-converting Enzyme Inhibitors Potentiate Preconditioning Througy Bradyikiik B2 Receptor Activation in Human Heart [J] J Am Coll Cardiol,1997,29:1599-1606.
[26]Witherow FN, Helmy A,Webb DJ,et al.Bradykinin contributes to the vasodilator effects of chronic angiotensin-converting enzyme inhibiton in patients with heart failer. Circulation,2001,104(18):2177-2181.
[27]M.C.Chappell,S.N.iyer.D.L.Diz et al. Antihypertensive effects of Angiotensin-(1-7).Braz J Med.Biol.Res,1998,31:1205-1212.
[28]Kucharewicz I,Pawlak R,Matys T.et al.Antithrombotic effect of captopril and losartan is mediated by angiotensin-(1-7).Hypertension,2002,40:774.
[29]FerreIR AJ,Santos RA,Almeida AP,Angiotension-(1-7):Cardioprotective effect in myocardial ischemia/reperfusion.Hypertension,2001,38:665:668.
[30]刘风芝,李延平黄明莉,等.冬虫夏草醇提取物对大鼠缺血再灌注过程心肌保护作用研究.中国病理生理杂志,1999;15(3)：240.
[31]郭向阳,徐守春,罗爱伦,等.卡托普利对心肌组织高能磷酸化合物代谢的影响中华胸心血管外科杂志,1998；14：115.