亚低温对垂体后叶素所致实验性大鼠心肌缺血的影响
摘要
[实验目的及意义]
早在50年代，人们就将深低温（<27℃）应用于心内直视手术，以保存重要脏器功能。 1978年，Abendschein等报告了人工低温（26℃）能明显缩小夹闭狗冠脉后5或10小时的心肌缺血损伤范围，使前者的MIS缩小25%，后者的MIS缩小近20%。但深低温使P50变小，氧离曲线左移，血红蛋白向组织释放氧量减少。浅低温（33-35℃）不明显干扰机体内稳态，它可以使心脏保持一定的机械电活动，有利于心肌细胞对氧的摄取和利用，从而有效地避免了低温所导致的酶活性受抑、线粒体功能受影响的过程，也避免了低温容易引起心室颤动，延长心脏复跳时间等不良后果。国内钱远宇等人研究了亚低温疗法对家兔持续性急性心肌梗死范围的影响，亚低温疗法缩小家兔持续性急性心肌梗死范围，增加濒危区的存活心肌。以往研究主要用开胸手术制备心梗模型后得到亚低温可以减少心梗面积的结论。开胸手术虽有梗死部位确切的优点，但容易导致气胸等手术副损伤，且实验动物容易死亡，手术技术要求较高；同时，它属于局部的心肌坏死，区别于整体的心肌缺血（global ischemia）。目前亚低温被广泛的应用于临床的心肺复苏和神经系统疾病的治疗，一部分患者不但有神经系统疾病同时还存在循环系统疾病，比如说由于身体广泛的动脉粥样硬化，患者可以在冠心病的基础上出现蛛网膜下腔出血、脑出血、脑梗塞等，
对于此类患者是否可以应用亚低温治疗，即亚低温对于整体的心肌缺血（global ischemia）的作用如何，目前尚无报道，本文采用垂体后叶素制备动物实验性心肌缺血模型，观察亚低温对大鼠实验性心肌缺血的影响，探讨其机理，并首次观察了亚低温对心功能的影响，为亚低温方法用于心肌缺血的临床治疗提供了试验依据。同时指出降温对心梗的治疗的重要性。
[实验方法]
Wistar大鼠50只，分为5组，每组10只，A组为常温+生理盐水+垂体后叶素腹腔注射：B组为常温+硝酸甘油+垂体后叶素腹腔注射：C组为低温+生理盐水+垂体后叶素腹腔注射；D组为低温+硝酸甘油+垂体后叶素腹腔注射;E组为假手术组。A、C、E组给予腹腔注射生理盐水1ml，B、D组给硝酸甘油5mg/kg体重腹腔注射；记录给药前及给药后30分钟心电图及给药前A、C组±dp/dtmax、LVSP。腹腔注射30分钟后，A、B、C、D组经腹腔注射垂体后叶素30u/kg，E组注射等量生理盐水记录注射后即刻、30秒、1分、5分、10分、15分、30分心电图；记录注射垂体后叶素后30分钟A、C组±dp/dtmax、LVSP。在整个实验过程中每只大鼠持续监测肛温。大鼠正常体温在36-37℃之间。A、B、E组保持体温在37℃以上。C、D两低温组将4℃生理盐水袋置于仰卧的大鼠体下进行缓慢的体表降温，避免产生寒战等反应，温度下降明显时给予电热毯及白炽灯照射复温，使肛温在垂体后叶素注射后保持在32-35℃之间。C组动物实验结束后缓慢复温至正常体温。对于A、C、E组实验动物24小时后正中开胸，取下心脏，用生理盐水洗去心脏中的残血，投入10%甲醛溶液中固定。固定后的标本进行石蜡包埋,切片后进行HE染色,在光镜下观察。
[实验结果]
心电图阴性率的比较
A、 B、C、D四组中低温盐水组C组的阴性率最高为80%，低温硝酸甘油组D组其次为70%，常温硝酸甘油组B组高于常温盐水组A组为60%。低温盐水组C组与常温对照A组比较差异显著P<0.01，D组、B组分别与A组比较P<0.05。C与B组、D组与B组比较无统计学意义。
心功能测定
垂体后叶素注射后常温组与低温组的+dp/dtmax均有所下降，常温组低温组给药30分钟后降至（5455.3±904.6 mmHg/s），常温组由（5735.0±1040.3 mmHg/s）下降至（4009.5±970.9 mmHg/s），两者差异显著，P<0.001。
垂体后叶素注射后常温组与低温组的-dp/dtmax也均有所下降，低温组降至（4331.4±669.5 mmHg/s），常温组由（4558.2±955.6 mmHg/s）下降至（3500.3±628.4 mmHg/s），但低温组下降的程度小于常温组，P＜0.001。
垂体后叶素注射30分钟后的常温组的LVSP与低温组的LVSP均有所下降，低温组降至（115.2±8.2 mmHg），常温组由（120.7±12.9 mmHg）下降至（84.3±11.8 mmHg），但低温组下降的程度小于常温组，P＜0.001。
HE染色
常温组阳性病理切片主要表现为心肌细胞有疏松化，肌纤维呈颗粒状（早期缺血改变）；低温组阳性病理切片主要表现为心外膜血管扩张充血，心肌细胞间水肿，局灶性心肌细胞疏松化。
[讨论]
实验结果表明亚低温增加了垂体后叶素所致实验性大鼠心肌缺血的阴性率,增加了缺血后大鼠的心肌收缩力和左室功能，病理上也减轻了大鼠心肌缺血的程度。其对硝酸甘油扩冠作用无明显影响。这些结果表明亚低温对垂体后叶素所致大鼠心肌缺血
有确切的保护作用。
降温后由于组织细胞降低了对代谢物的需求，使得对这些物质供应的依赖性减少。当代谢率很低或停止时，组织的活性可以延长。这一效应说明，降温能够防止缺血的有害作用。组织的降温可以保存其能量贮备。低温至33-30℃以下时，可减轻心肌缺血时膜的通透性异常升高，使细胞内钙离子的积聚减轻，肌酸激酶和α-羟丁酸脱氢酶的逸出明显减少，细胞膜的正常通透性得到明显的保存。人工低温可降低细胞的代谢率，减少酸?
Objectives As early as 50's, people utilized the profound hypothermia(<27 ℃ ) in the open heart operation, to preserve the function of important organs . In 1978, Abendschein etc. reported that the artificial cooling(26 ℃ ) could obviously contract the myocardial infarction size after 5 or 10 hours coronary artery occlusion of dog ,contracting the former MIS 25%, the MIS of the latter contracted near 20%.But the profound hypothermia make P50 diminished, the oxygen -leave- curve shift left, the oxygen capacity that hemoglobin releases to tissues decrease. The mild hypothermia(33-35 ℃ ) do not interfere the hemostasis obviously, it can make the heart to keep the certain machine electricity activity, benefitting to the myocardial cell to take in the oxygen and make use of it . So it avoided availably the activity of enzyme from restraining by the low temperature, and the influence of the function in the mitochondrion. It also avoided the ventricular fibrillation that caused by the low temperature, prolonging to recover jumping, and the bad results in etc. In our country Qian etc. studied the effect of mild hypothermia on the myocardial infarction size in continuous coronary artery occluded rabbit. Mild hypothermia might reduce infarction size in the rabbits with continuous coronary artery occluded acute myocardial infarction, and increase survied myocardium in the risk zone. The former research got the conclusion that mild hypothermia could reduce the myocardial infarction size using open-chest-surgical operation. Although it has the merit that the scope of myocardial infarction is sure, open- chest-surgical-operation causes aerothorax easily as vice- harm. Animal would die easily. The surgical operation technique request is higher. At the same time, it belongs to the partial myocardial necrosis, and is different from the global ischemia.
At present mild hypothermia is used in the treatment of the nervous system disease and
cardiopulmonary resuscitation. Parts of the sufferers not only have the nervous system disease, but also have something wrong with the circulatory system. For example, because of the extensive atherosclerosis in the body, based on the coronary heart disease people might be sufferered by the hemorrhage of subarachniod space, apoplexy, and cerebral inarction etc. For this kind of sufferer whether can apply the mild hypothermia cure, namely the effect of mild hypothermia on the global ischemia is uncertain. The object of our study is to observe the effect of mild hypothermia on the myocardial ischemia caused by Pituitrin, investigate the underlying mechanism, and provide experiment basis to the method of mild hypothermia as a clinical treatment for myocardial ischemia.
Methods 50 Wistar rats were randomized to five groups: group E of sham operation, myocardial ischemia group A with normothermia, myocardial ischemia group C with hypothermia, myocardial ischemia group B treated with Nitroglycerin under normal temperature, and myocardial ischemia group D treated with Nitroglycerin under low temperature, each group has 10 rats. The A, C, E groups were treated with Saline 1ml belly injecting; group B, D treated with Nitroglycerin (5mg/kg). 30 minutes later Pit (30U/kg) was given to each group except for the group E. Record the ECG ,and LVSP ±dp/dtmax of group A and C. Core temperature was measured with thermistor. The mild hypothermia group received cooling under the body with the 4 ℃ Saline bag. The core temperature was dropped to 32-35℃, while the control group’s body temperature was kept above 37℃. Afterwards, hearts were excised and cut to several sections stained with HE for observing the degree of myocardial ischemia.
Results 1.Comparison of the electrocardiogram negative rate The negative rate of hypothermia group C was the most high for 80% which was compared with A, B, D groups. The negative rate of group D treated with Nitroglycerin under low temperature was the second. Group B treated with Nitroglycerin under normal temperature was higher than group A without
早在50年代，人们就将深低温（<27℃）应用于心内直视手术，以保存重要脏器功能。 1978年，Abendschein等报告了人工低温（26℃）能明显缩小夹闭狗冠脉后5或10小时的心肌缺血损伤范围，使前者的MIS缩小25%，后者的MIS缩小近20%。但深低温使P50变小，氧离曲线左移，血红蛋白向组织释放氧量减少。浅低温（33-35℃）不明显干扰机体内稳态，它可以使心脏保持一定的机械电活动，有利于心肌细胞对氧的摄取和利用，从而有效地避免了低温所导致的酶活性受抑、线粒体功能受影响的过程，也避免了低温容易引起心室颤动，延长心脏复跳时间等不良后果。国内钱远宇等人研究了亚低温疗法对家兔持续性急性心肌梗死范围的影响，亚低温疗法缩小家兔持续性急性心肌梗死范围，增加濒危区的存活心肌。以往研究主要用开胸手术制备心梗模型后得到亚低温可以减少心梗面积的结论。开胸手术虽有梗死部位确切的优点，但容易导致气胸等手术副损伤，且实验动物容易死亡，手术技术要求较高；同时，它属于局部的心肌坏死，区别于整体的心肌缺血（global ischemia）。目前亚低温被广泛的应用于临床的心肺复苏和神经系统疾病的治疗，一部分患者不但有神经系统疾病同时还存在循环系统疾病，比如说由于身体广泛的动脉粥样硬化，患者可以在冠心病的基础上出现蛛网膜下腔出血、脑出血、脑梗塞等，
对于此类患者是否可以应用亚低温治疗，即亚低温对于整体的心肌缺血（global ischemia）的作用如何，目前尚无报道，本文采用垂体后叶素制备动物实验性心肌缺血模型，观察亚低温对大鼠实验性心肌缺血的影响，探讨其机理，并首次观察了亚低温对心功能的影响，为亚低温方法用于心肌缺血的临床治疗提供了试验依据。同时指出降温对心梗的治疗的重要性。
[实验方法]
Wistar大鼠50只，分为5组，每组10只，A组为常温+生理盐水+垂体后叶素腹腔注射：B组为常温+硝酸甘油+垂体后叶素腹腔注射：C组为低温+生理盐水+垂体后叶素腹腔注射；D组为低温+硝酸甘油+垂体后叶素腹腔注射;E组为假手术组。A、C、E组给予腹腔注射生理盐水1ml，B、D组给硝酸甘油5mg/kg体重腹腔注射；记录给药前及给药后30分钟心电图及给药前A、C组±dp/dtmax、LVSP。腹腔注射30分钟后，A、B、C、D组经腹腔注射垂体后叶素30u/kg，E组注射等量生理盐水记录注射后即刻、30秒、1分、5分、10分、15分、30分心电图；记录注射垂体后叶素后30分钟A、C组±dp/dtmax、LVSP。在整个实验过程中每只大鼠持续监测肛温。大鼠正常体温在36-37℃之间。A、B、E组保持体温在37℃以上。C、D两低温组将4℃生理盐水袋置于仰卧的大鼠体下进行缓慢的体表降温，避免产生寒战等反应，温度下降明显时给予电热毯及白炽灯照射复温，使肛温在垂体后叶素注射后保持在32-35℃之间。C组动物实验结束后缓慢复温至正常体温。对于A、C、E组实验动物24小时后正中开胸，取下心脏，用生理盐水洗去心脏中的残血，投入10%甲醛溶液中固定。固定后的标本进行石蜡包埋,切片后进行HE染色,在光镜下观察。
[实验结果]
心电图阴性率的比较
A、 B、C、D四组中低温盐水组C组的阴性率最高为80%，低温硝酸甘油组D组其次为70%，常温硝酸甘油组B组高于常温盐水组A组为60%。低温盐水组C组与常温对照A组比较差异显著P<0.01，D组、B组分别与A组比较P<0.05。C与B组、D组与B组比较无统计学意义。
心功能测定
垂体后叶素注射后常温组与低温组的+dp/dtmax均有所下降，常温组低温组给药30分钟后降至（5455.3±904.6 mmHg/s），常温组由（5735.0±1040.3 mmHg/s）下降至（4009.5±970.9 mmHg/s），两者差异显著，P<0.001。
垂体后叶素注射后常温组与低温组的-dp/dtmax也均有所下降，低温组降至（4331.4±669.5 mmHg/s），常温组由（4558.2±955.6 mmHg/s）下降至（3500.3±628.4 mmHg/s），但低温组下降的程度小于常温组，P＜0.001。
垂体后叶素注射30分钟后的常温组的LVSP与低温组的LVSP均有所下降，低温组降至（115.2±8.2 mmHg），常温组由（120.7±12.9 mmHg）下降至（84.3±11.8 mmHg），但低温组下降的程度小于常温组，P＜0.001。
HE染色
常温组阳性病理切片主要表现为心肌细胞有疏松化，肌纤维呈颗粒状（早期缺血改变）；低温组阳性病理切片主要表现为心外膜血管扩张充血，心肌细胞间水肿，局灶性心肌细胞疏松化。
[讨论]
实验结果表明亚低温增加了垂体后叶素所致实验性大鼠心肌缺血的阴性率,增加了缺血后大鼠的心肌收缩力和左室功能，病理上也减轻了大鼠心肌缺血的程度。其对硝酸甘油扩冠作用无明显影响。这些结果表明亚低温对垂体后叶素所致大鼠心肌缺血
有确切的保护作用。
降温后由于组织细胞降低了对代谢物的需求，使得对这些物质供应的依赖性减少。当代谢率很低或停止时，组织的活性可以延长。这一效应说明，降温能够防止缺血的有害作用。组织的降温可以保存其能量贮备。低温至33-30℃以下时，可减轻心肌缺血时膜的通透性异常升高，使细胞内钙离子的积聚减轻，肌酸激酶和α-羟丁酸脱氢酶的逸出明显减少，细胞膜的正常通透性得到明显的保存。人工低温可降低细胞的代谢率，减少酸?
Objectives As early as 50's, people utilized the profound hypothermia(<27 ℃ ) in the open heart operation, to preserve the function of important organs . In 1978, Abendschein etc. reported that the artificial cooling(26 ℃ ) could obviously contract the myocardial infarction size after 5 or 10 hours coronary artery occlusion of dog ,contracting the former MIS 25%, the MIS of the latter contracted near 20%.But the profound hypothermia make P50 diminished, the oxygen -leave- curve shift left, the oxygen capacity that hemoglobin releases to tissues decrease. The mild hypothermia(33-35 ℃ ) do not interfere the hemostasis obviously, it can make the heart to keep the certain machine electricity activity, benefitting to the myocardial cell to take in the oxygen and make use of it . So it avoided availably the activity of enzyme from restraining by the low temperature, and the influence of the function in the mitochondrion. It also avoided the ventricular fibrillation that caused by the low temperature, prolonging to recover jumping, and the bad results in etc. In our country Qian etc. studied the effect of mild hypothermia on the myocardial infarction size in continuous coronary artery occluded rabbit. Mild hypothermia might reduce infarction size in the rabbits with continuous coronary artery occluded acute myocardial infarction, and increase survied myocardium in the risk zone. The former research got the conclusion that mild hypothermia could reduce the myocardial infarction size using open-chest-surgical operation. Although it has the merit that the scope of myocardial infarction is sure, open- chest-surgical-operation causes aerothorax easily as vice- harm. Animal would die easily. The surgical operation technique request is higher. At the same time, it belongs to the partial myocardial necrosis, and is different from the global ischemia.
At present mild hypothermia is used in the treatment of the nervous system disease and
cardiopulmonary resuscitation. Parts of the sufferers not only have the nervous system disease, but also have something wrong with the circulatory system. For example, because of the extensive atherosclerosis in the body, based on the coronary heart disease people might be sufferered by the hemorrhage of subarachniod space, apoplexy, and cerebral inarction etc. For this kind of sufferer whether can apply the mild hypothermia cure, namely the effect of mild hypothermia on the global ischemia is uncertain. The object of our study is to observe the effect of mild hypothermia on the myocardial ischemia caused by Pituitrin, investigate the underlying mechanism, and provide experiment basis to the method of mild hypothermia as a clinical treatment for myocardial ischemia.
Methods 50 Wistar rats were randomized to five groups: group E of sham operation, myocardial ischemia group A with normothermia, myocardial ischemia group C with hypothermia, myocardial ischemia group B treated with Nitroglycerin under normal temperature, and myocardial ischemia group D treated with Nitroglycerin under low temperature, each group has 10 rats. The A, C, E groups were treated with Saline 1ml belly injecting; group B, D treated with Nitroglycerin (5mg/kg). 30 minutes later Pit (30U/kg) was given to each group except for the group E. Record the ECG ,and LVSP ±dp/dtmax of group A and C. Core temperature was measured with thermistor. The mild hypothermia group received cooling under the body with the 4 ℃ Saline bag. The core temperature was dropped to 32-35℃, while the control group’s body temperature was kept above 37℃. Afterwards, hearts were excised and cut to several sections stained with HE for observing the degree of myocardial ischemia.
Results 1.Comparison of the electrocardiogram negative rate The negative rate of hypothermia group C was the most high for 80% which was compared with A, B, D groups. The negative rate of group D treated with Nitroglycerin under low temperature was the second. Group B treated with Nitroglycerin under normal temperature was higher than group A without
引文
高绪孟，低温医学，第一版 北京 人民卫生出版社 1991，15-16。
王祥瑞,陈长志，电针刺激辅助低温对猪缺血的影响[J], 中国中西医结合急救杂志2001年3月第8卷第2期,75-76
王宪等，人工低温对大鼠实验性心肌梗塞的影响，I对心肌梗塞范围和血液动力学的影响[J]，北京医科大学学报 1987；19（5）：319
李广斌，郑从义， 唐兵，低温生物学 ，第一版 长沙 湖南科学技术出版社，1998， 121-123。
钱远宇,孟庆义，王志忠，马勇，亚低温疗法缩小家兔持续性急性心肌梗塞范围的研究[J]，中国急救医学2003，7，23（7）：449-451。
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Chien GL ,wolff RA . Davis RF .Van Winkle DM. Normothermicrage temperature affects myocardial infact size Cardiovase Res 1994; 28:1014-1017.
Mike T , Cohen MV , Downey JM . Hypothermia’s cardioprotective effect is additive to that of ischemic preconditioning Circulation 1997;96(1):687.
Miki T; Liu GS; Cohen MV; Downey JM. Mild hypothermia reduces infarct size in the beating rabbit heart: a practical intervention for acute myocardial infarction?[J] Basic research in cardiology [Basic Res Cardiol] 1998 Oct; 93 (5), pp. 372-83.
王志忠，孟庆义，亚低温对兔心肌血预处理后急性心肌梗死面积的影响[J]，中华老年心脑血管病杂志， 2003， 5（5），353。
Sheldrick A; Gray KM; Drew GM; Louttit JB. The effect of body temperature on myocardial protection conferred by ischaemic preconditioning or the selective
adenosine A1 receptor agonist GR79236, in an anaesthetized rabbit model of myocardial ischaemia and reperfusion.[J] British journal of pharmacology [Br J Pharmacol] 1999 Sep; 128 (2), pp. 385-95.
王德生等，亚低温脑保护作用，黑龙江，科技出版社，2000，7
Hale SL; Kloner RA. Myocardial temperature reduction attenuates necrosis after prolonged ischemia in rabbits.[J] Cardiovascular research [Cardiovasc Res] 1998 Dec; 40 (3), pp. 502-7.
Hale SL; Kloner RA. Myocardial temperature in acute myocardial infarction: protection with mild regional hypothermia.[J] American journal of physiology [Am J Physiol] 1997 Jul; 273 (1 Pt 2), pp. H220-7.
郭鹞编，人类疾病的动物模型，北京，人民卫生出版社 1982 ：367。
谢人明、李胜涛，益气通脉口服液抗试验性大鼠心肌缺血的实验研究[J]，首都中医药大学学报2001年12月第24卷第4期，43-44。
Black JW .Electrocardiographic changes produced in rabbits by vasopresin(pitressin) and their alterations by prolonged treament with a conamercial heat extract .J Pharma Pharmacol 1960; 12:87.
吴伟康，杨仕云，心肌缺血时NO代谢的动态变化，中国病理杂志2002,18(10):1258-1261。
Klein K , Snan D . Effect of cold on muscled potential and etetrolytes . Metabllism , 1968, 27: 1094-1098。
Ning XH; Xu CS; Song YC; Xiao Y; Hu YJ; Lupinetti FM; Portman MA. Temperature threshold and preservation of signaling for mitochondrial membrane proteins during ischemia in rabbit heart. [J] Cryobiology [Cryobiology] 1998 Jun; 36 (4), pp. 321-9.
Ning XH; Xu CS; Song YC; Childs KF; Xiao Y; Bolling SF; Lupinetti FM; Portman MA.Temperature threshold and modulation of energy metabolism in the cardioplegic arrested rabbit heart.[J] Cryobiology [Cryobiology] 1998 Feb; 36 (1), pp. 2-11.
Antman EM, Braunwald E. Acute myocardial infarction. In: Braunwald E, editor.
Heart disease. A textbook of cardiovascular medicine. Philadelphia: Saunders; 1997. p. 1200.
Lazenby WE,KOW ,Zelano JA.Effect of temperature and flow rate on regional blood and metabolism during cardiopulmonary by pass.Am Thorac Surg,1992,53(6):957-962.
Hale SL, Dave RH, Kloner RA. Regional hypothermia reduces myocardial necrosis even when instituted after the onset of ischemia. Basic Res Cardiol 1997; 92:351–357.
YellonDM,PasiniE,CargnoniA,etal.Theprotectroleofheartstressintheischemicandreperfusedrabbitmyocardium.JMolCellCardiol,1992,24:895-902
Yamauchi T; Ichikawa H; Sawa Y; Fukushima N; Kagisaki K; Maeda K; Matsuda H; Shirakura RThe contribution of Na+/H+ exchange to ischemia-reperfusion injury after hypothermic cardioplegic arrest.[J] Annals of thoracic surgery [Ann Thorac Surg] 1997 Apr; 63 (4), pp. 1107-12.
Hearse DJ , et al The oxygen paradox and the calcium paradox . Two facts of same Problem ? J Mol cell Cardiol 1978; 10: 641.
Strother SV, Bull JM, Branham SA. Activation of coagulation during therapeutic whole body hyperthermia. Thromb Res 1986; 43:353–360.
Gray CC; Amrani M; Yacoub MH.Heat stress proteins and myocardial protection: experimental model or potential clinical tool?[J] international journal of biochemistry & cell biology [Int J Biochem Cell Biol] 1999 May; 31 (5), pp. 559-73.
Chayen J,et al.Practical histochemistry Londen, New York，John Wiley and Sons, 1973:197 .127.36.
BalerR,DahlG,VoellmyR. Activation of human heart shock genesis accompanied byoligomerization ,modification,and rapid translocation of heart shock transcription factor HSF1.MolCellBiol,1993,13:2486-2493.
Currie RW, Tanguay RM, Kingma JG, Jr. Heat-shock response and limitation of tissue necrosis during occlusion/reperfusion in rabbit hearts. Circulation 1993; 87:963–971.
Inoue K; Ando S; Gyuan F; Takaba T A study of the myocardial protective effect of rapid cooling based on intracellular Ca, intracellular pH, and HSP70. [J] Annals of thoracic and cardiovascular surgery : official journal of the Association of Thoracic and Cardiovascular Surgeons of Asia [Ann Thorac Cardiovasc Surg] 2003 Oct; 9 (5), pp. 301-6.
Yamashita N, Hoshida S, Nishida M, Igarashi J, Aoki K, Hori M, et al. Time course of tolerance to ischemia-reperfusion injury and induction of heat shock protein 72 by heat stress in the rat heart. J Mol Cell Cardiol 1997; 29:1815–1821.
Yamashita N, Hoshida S, Taniguchi N, Kuzuya T, Hori M. Whole-body hyperthermia provides biphasic cardioprotection against ischemia/reperfusion injury in rats. Circulation 1998; 98:1414–1421.
杨彦玲，师养荣，心肌缺血再灌注损伤研究进展[J]，心血管病学进展2003年第24卷第2期，116-120
Corliss Rj ,et al. systemic and coronary hemodynamic effects of vasopressin . Am J Med sci 1968; 256:293.
解启莲，胡浩天，安会波，亚低温对全身炎症反应小鼠细胞因子与粘附分子表达的影响及意义[J]，中国危重症急救医学， 2000 年12（10），715-716。
王祥瑞,陈长志，电针刺激辅助低温对猪缺血的影响[J], 中国中西医结合急救杂志2001年3月第8卷第2期,75-76
王宪等，人工低温对大鼠实验性心肌梗塞的影响，I对心肌梗塞范围和血液动力学的影响[J]，北京医科大学学报 1987；19（5）：319
李广斌，郑从义， 唐兵，低温生物学 ，第一版 长沙 湖南科学技术出版社，1998， 121-123。
钱远宇,孟庆义，王志忠，马勇，亚低温疗法缩小家兔持续性急性心肌梗塞范围的研究[J]，中国急救医学2003，7，23（7）：449-451。
Hale SL; Kloner RA. Elevated body temperature during myocardial ischemia/reperfusion exacerbates necrosis and worsens no-reflow.[J] Coronary artery disease [Coron Artery Dis] 2002 May; 13 (3), pp. 177-81.
Duncker DJ , Klassen CL , Ishibashi Y ,et al, Effect of temperature on myocardial in swine . Am J Physiol 1996; 270:H1189-h1199.
Chien GL ,wolff RA . Davis RF .Van Winkle DM. Normothermicrage temperature affects myocardial infact size Cardiovase Res 1994; 28:1014-1017.
Mike T , Cohen MV , Downey JM . Hypothermia’s cardioprotective effect is additive to that of ischemic preconditioning Circulation 1997;96(1):687.
Miki T; Liu GS; Cohen MV; Downey JM. Mild hypothermia reduces infarct size in the beating rabbit heart: a practical intervention for acute myocardial infarction?[J] Basic research in cardiology [Basic Res Cardiol] 1998 Oct; 93 (5), pp. 372-83.
王志忠，孟庆义，亚低温对兔心肌血预处理后急性心肌梗死面积的影响[J]，中华老年心脑血管病杂志， 2003， 5（5），353。
Sheldrick A; Gray KM; Drew GM; Louttit JB. The effect of body temperature on myocardial protection conferred by ischaemic preconditioning or the selective
adenosine A1 receptor agonist GR79236, in an anaesthetized rabbit model of myocardial ischaemia and reperfusion.[J] British journal of pharmacology [Br J Pharmacol] 1999 Sep; 128 (2), pp. 385-95.
王德生等，亚低温脑保护作用，黑龙江，科技出版社，2000，7
Hale SL; Kloner RA. Myocardial temperature reduction attenuates necrosis after prolonged ischemia in rabbits.[J] Cardiovascular research [Cardiovasc Res] 1998 Dec; 40 (3), pp. 502-7.
Hale SL; Kloner RA. Myocardial temperature in acute myocardial infarction: protection with mild regional hypothermia.[J] American journal of physiology [Am J Physiol] 1997 Jul; 273 (1 Pt 2), pp. H220-7.
郭鹞编，人类疾病的动物模型，北京，人民卫生出版社 1982 ：367。
谢人明、李胜涛，益气通脉口服液抗试验性大鼠心肌缺血的实验研究[J]，首都中医药大学学报2001年12月第24卷第4期，43-44。
Black JW .Electrocardiographic changes produced in rabbits by vasopresin(pitressin) and their alterations by prolonged treament with a conamercial heat extract .J Pharma Pharmacol 1960; 12:87.
吴伟康，杨仕云，心肌缺血时NO代谢的动态变化，中国病理杂志2002,18(10):1258-1261。
Klein K , Snan D . Effect of cold on muscled potential and etetrolytes . Metabllism , 1968, 27: 1094-1098。
Ning XH; Xu CS; Song YC; Xiao Y; Hu YJ; Lupinetti FM; Portman MA. Temperature threshold and preservation of signaling for mitochondrial membrane proteins during ischemia in rabbit heart. [J] Cryobiology [Cryobiology] 1998 Jun; 36 (4), pp. 321-9.
Ning XH; Xu CS; Song YC; Childs KF; Xiao Y; Bolling SF; Lupinetti FM; Portman MA.Temperature threshold and modulation of energy metabolism in the cardioplegic arrested rabbit heart.[J] Cryobiology [Cryobiology] 1998 Feb; 36 (1), pp. 2-11.
Antman EM, Braunwald E. Acute myocardial infarction. In: Braunwald E, editor.
Heart disease. A textbook of cardiovascular medicine. Philadelphia: Saunders; 1997. p. 1200.
Lazenby WE,KOW ,Zelano JA.Effect of temperature and flow rate on regional blood and metabolism during cardiopulmonary by pass.Am Thorac Surg,1992,53(6):957-962.
Hale SL, Dave RH, Kloner RA. Regional hypothermia reduces myocardial necrosis even when instituted after the onset of ischemia. Basic Res Cardiol 1997; 92:351–357.
YellonDM,PasiniE,CargnoniA,etal.Theprotectroleofheartstressintheischemicandreperfusedrabbitmyocardium.JMolCellCardiol,1992,24:895-902
Yamauchi T; Ichikawa H; Sawa Y; Fukushima N; Kagisaki K; Maeda K; Matsuda H; Shirakura RThe contribution of Na+/H+ exchange to ischemia-reperfusion injury after hypothermic cardioplegic arrest.[J] Annals of thoracic surgery [Ann Thorac Surg] 1997 Apr; 63 (4), pp. 1107-12.
Hearse DJ , et al The oxygen paradox and the calcium paradox . Two facts of same Problem ? J Mol cell Cardiol 1978; 10: 641.
Strother SV, Bull JM, Branham SA. Activation of coagulation during therapeutic whole body hyperthermia. Thromb Res 1986; 43:353–360.
Gray CC; Amrani M; Yacoub MH.Heat stress proteins and myocardial protection: experimental model or potential clinical tool?[J] international journal of biochemistry & cell biology [Int J Biochem Cell Biol] 1999 May; 31 (5), pp. 559-73.
Chayen J,et al.Practical histochemistry Londen, New York，John Wiley and Sons, 1973:197 .127.36.
BalerR,DahlG,VoellmyR. Activation of human heart shock genesis accompanied byoligomerization ,modification,and rapid translocation of heart shock transcription factor HSF1.MolCellBiol,1993,13:2486-2493.
Currie RW, Tanguay RM, Kingma JG, Jr. Heat-shock response and limitation of tissue necrosis during occlusion/reperfusion in rabbit hearts. Circulation 1993; 87:963–971.
Inoue K; Ando S; Gyuan F; Takaba T A study of the myocardial protective effect of rapid cooling based on intracellular Ca, intracellular pH, and HSP70. [J] Annals of thoracic and cardiovascular surgery : official journal of the Association of Thoracic and Cardiovascular Surgeons of Asia [Ann Thorac Cardiovasc Surg] 2003 Oct; 9 (5), pp. 301-6.
Yamashita N, Hoshida S, Nishida M, Igarashi J, Aoki K, Hori M, et al. Time course of tolerance to ischemia-reperfusion injury and induction of heat shock protein 72 by heat stress in the rat heart. J Mol Cell Cardiol 1997; 29:1815–1821.
Yamashita N, Hoshida S, Taniguchi N, Kuzuya T, Hori M. Whole-body hyperthermia provides biphasic cardioprotection against ischemia/reperfusion injury in rats. Circulation 1998; 98:1414–1421.
杨彦玲，师养荣，心肌缺血再灌注损伤研究进展[J]，心血管病学进展2003年第24卷第2期，116-120
Corliss Rj ,et al. systemic and coronary hemodynamic effects of vasopressin . Am J Med sci 1968; 256:293.
解启莲，胡浩天，安会波，亚低温对全身炎症反应小鼠细胞因子与粘附分子表达的影响及意义[J]，中国危重症急救医学， 2000 年12（10），715-716。