Functional and metabolic protection of the neonatal myocardium from ischemia. Insufficient protection by cardioplegia

The effects of ischemia and cardiac arrest by cardioplegia on the mechanical function and energy metabolism of the ventricular myocardium of the neonatal guinea pig were investigated in the isolated perfused heart preparation and compared with these effects in the adult guinea pig. Whereas reperfusion after ischemia resulted in better recovery of mechanical function and a higher adenosine triphosphase content in the neonatal myocardium than in the adult, recovery from cardiac arrest induced by St. Thomas' Hospital cardioplegic solution was not as good in the neonatal myocardium as in the adult. Contracture developed in the neonatal myocardium on administration of the cardioplegic solution, but did not in the adult. This was considered to be the reason that the protective effect of the cardioplegic solution was inferior in the neonatal myocardium to that in the adult.     

Protection of the myocardium during global ischemia. Is crystalloid cardioplegia effective in the immature myocardium?

In pediatric cardiac operations, a high proportion of hospital deaths are believed to result from inadequate myocardial protection during the period of global ischemia. To investigate whether this may be due to an inherently lower resistance to myocardial ischemia or to the failure of conventional cardioplegia to afford adequate protection in the immature heart, we have conducted a series of studies with isolated hearts from neonatal (3 to 5 days old, body weight 6.3 to 13.4 gm) and adult (84 to 112 days old, 260 to 340 gm) rats. The efficacy of cardioplegia was assessed in neonatal hearts (n = 6 per group) subjected to various durations of normothermic ischemia, with and without a 2-minute preischemic infusion of the St. Thomas' Hospital cardioplegic solution. At all times studied, the use of cardioplegia resulted in a greater postischemic recovery of left ventricular developed pressure and first derivative of left ventricular pressure. After periods of ischemia lasting 30, 60, 90, 120, and 150 minutes in the absence of cardioplegia, left ventricular developed pressure recovered to 80% +/- 10%, 66% +/- 11%, 53% +/- 7%, 33% +/- 6%, and 21% +/- 4% of preischemic values, respectively; in the presence of cardioplegia, the values were 89% +/- 6%, 83% +/- 8%, 74% +/- 6% (p less than 0.05), 58% +/- 5% (p less than 0.05), and 41% +/- 7% (p less than 0.05), respectively. The corresponding values for first derivative of left ventricular pressure were 78% +/- 9%, 67% +/- 12%, 54% +/- 7%, 30% +/- 5%, and 19% +/- 3% in the absence of cardioplegia and 92% +/- 7%, 88% +/- 8%, 75% +/- 8%, 56% +/- 5% (p less than 0.05) and 39% +/- 6% (p less than 0.05) in the presence of cardioplegia. In the noncardioplegia groups, 90% of hearts exhibited ischemic contracture (mean time to onset = 24.7 +/- 1.1 minutes), whereas in the cardioplegia groups, only 63% exhibited contracture, and of a significantly delayed onset (37.0 +/- 1.5 min, p less than 0.05). Adult hearts (n = 5) subjected to 30 minutes of normothermic ischemic arrest, in the absence of cardioplegia, recovered 36% +/- 7% of the preischemic left ventricular developed pressure and 37% +/- 9% of the preischemic first derivative of left ventricular pressure on reperfusion; 100% of these hearts exhibited some degree of contracture (mean time to onset = 15.4 +/- 1.1 minutes) by the end of the ischemic period.(ABSTRACT TRUNCATED AT 400 WORDS)     

Studies of myocardial protection in the immature heart. I. Enhanced tolerance of immature versus adult myocardium to global ischemia with reference to metabolic differences

This study compares the metabolism and functional responses of adult and immature hearts to a standard ischemic insult. Ten adult dogs (25 to 27 kg) and 10 puppies (6 to 10 weeks old) underwent 45 minutes of aortic clamping on bypass. Preoperative and postoperative ventricular performance (Starling curves), biochemical factors, and water content were measured. Global ischemia in adults produced a 30% mortality rate (3/10) and low output syndrome in survivors (33% recovery of stroke work index). Conversely, all puppies survived and stroke work index returned to 85% of control, with less edema developing (0.4% versus 2% water gain, p less than 0.05). Puppies expended comparable glycogen stores but used more glutamate (15.4 versus 8.6 mumol/gm dry weight), produced more alanine (18.9 versus 6.4 mumol, p less than 0.05), succinate (19 versus 8.2 mumol, p less than 0.05), and malate (2.6 versus 0.15 mumol, p less than 0.05) during ischemia, and recovered better postischemic aerobic metabolism (410 versus 255 nmol tissue pyruvate, p less than 0.05). We conclude that tolerance of immature hearts to ischemia is related to amino acid utilization by transamination and increased substrate level phosphorylation, as occurring in diving mammals, suggesting retention of intrautero adaptive mechanisms.     

Enhanced myocardial protection during global ischemia with 5'-nucleotidase inhibitors.

Depletion of adenosine triphosphate precursors, such as myocardial adenosine, during global ischemia results in poor postischemic adenosine triphosphate repletion and functional recovery. Neonatal hearts may be more resistant to this deleterious effect of ischemia, because they are characterized by low 5'-nucleotidase activity, which may result in higher sustained endogenous myocardial adenosine triphosphate precursor levels during ischemia. Adult hearts, however, have high levels of 5'-nucleotidase activity leading to depleted precursors during ischemia and poor postischemic functional recovery. Augmenting myocardial adenosine exogenously during ischemia in adult hearts has a beneficial effect on recovery. The present study tested if preservation of nucleotide precursors, better adenosine triphosphate repletion, and enhanced postischemic myocardial recovery in adult hearts could be achieved with a "neonatal" strategy. Therefore 5'-nucleotidase inhibitors were administered to isolated, perfused adult rabbit hearts subjected to 120 minutes of ischemia (at 34 degrees C) to determine if this improved functional recovery. Hearts received St. Thomas' Hospital cardioplegic solution (control hearts) or cardioplegic solution containing 5'-nucleotidase inhibitors: pentoxifylline, thioinosine, [s-(p-nitrophenyl)-4-thioinosine], or thioinosine's dimethyl sulfoxide vehicle alone. After ischemia and reperfusion, recovery of systolic function, diastolic function, and myocardial oxygen consumption was significantly better with 5'-nucleotidase inhibition. No changes in coronary flow were noted. We speculate and are pursuing the theory that the mechanism of 5'-nucleotidase inhibition's favorable action is due to preventing the catabolism, transport, and loss of nucleotide precursors during ischemia, maintaining adenosine triphosphate precursor availability.     

Improved myocardial protection by aprotinin pretreatment in prolonged global ischemia

Protective effect of aprotinin pretreatment was assessed by functional, biochemical and morphological preservation in four hour global ischemia followed by one hour reperfusion in dogs. Cardioplegia was induced by intermittent infusion of cold Mg-lidocaine solution. Aprotinin 10,000 KIU/kg was given in low dose group (8 dogs), and 20,000 KIU/kg in high dose group (6 dogs); one half was given before ischemia and another half during ischemia. Betamethasone, coenzyme Q and nifedipine were also given equally in both groups before ischemia. Results were as follows: 1. Four (50%) of low dose group and all of high dose group were successfully taken off CPB and survived for one hour reperfusion. 2. High dose group showed significantly higher blood pressure and LVSWI than low dose group after one hour reperfusion (p less than 0.05). 3. Serum N-acetyl-beta-D-glucosaminidase and mitochondrial aspartate aminotransferase showed the significantly lower activity in high dose group than in low dose group after one hour reperfusion (p less than 0.05). There was no significant difference in the activities of serum beta-glucuronidase and MB-creatine kinase. 4. Myocardial tissues, excised after one hour reperfusion, contained significantly higher creatine phosphate in high dose group than in low dose group (p less than 0.05). There was no significant difference in the contents of adenosine triphosphate, calcium and water. 5. Severely injured mitochondrion were significantly lesser in high dose group than in low dose group. All lysosomes showed mild swelling or enlargement, but those membranous structures were well-preserved in both groups. In conclusion, aprotinin pretreatment might be effective in myocardial protection against prolonged global ischemia, by inhibiting the "leak out" of lysosomal enzymes.     

The protective effect of vitamin E on cerebral ischemia

Using the "canine model of the completely ischemic brain regulated with a perfusion method," the effects on cerebral ischemia of vitamin E, which is known to act as an antioxidant, were investigated. After pretreatment with vitamin E by oral or intravenous administration, cerebral blood flow was reduced to 1/10th the normal state and, 1 hour later, allowed to return to normal. Subsequent changes in electrical activity were observed, and the effects of vitamin E were evaluated. In the control group, no recovery of electrical activity was seen. In the groups given vitamin E, the recovery time was significantly shortened in the dogs given 30 mg/kg of vitamin E intravenously. Furthermore, in the groups treated with vitamin E, distinct recovery of electroencephalographic potentials at 3 hours after recirculation was apparent. These effects were more favorable in the case of intravenous administration than in the case of oral administration. These experimental results indicate that the administration of vitamin E is effective in protecting the brain from cerebral ischemia.     

Protection of the immature myocardium during global ischemia. A comparison of four clinical cardioplegic solutions in the rabbit heart

Controversy surrounds the reported beneficial effects of crystalloid cardioplegic solutions in the immature myocardium. In the present study we have investigated the efficacy of four clinical cardioplegic solutions in the immature myocardium to determine (1) whether cardioplegic protection could be demonstrated and, if so, (2) the relative efficacy of the four solutions. Isolated, working hearts (n = 6 per group) from neonatal rabbits (aged 5 to 8 days) were perfused aerobically (37 degrees C) for 20 minutes before a 2-minute infusion of one of four cardioplegic solutions: The St. Thomas' Hospital No. 2, Tyers, Bretschneider, and Roe solutions. Hearts were then rendered globally ischemic for 50 minutes at 37 degrees C before reperfusion for 15 minutes in the Langendorff mode and 20 minutes in the working mode. The postischemic recovery of cardiac function and leakage of creatine kinase were compared with results in noncardioplegic control hearts. Good protection was observed with the St. Thomas' Hospital and Tyers solutions: The postischemic recovery of cardiac output was increased from 21.2% +/- 12.7% in the cardioplegia-free group to 79.4% +/- 6.2% and 72.9% +/- 4.4%, respectively, in the St. Thomas' Hospital and Tyers groups (p less than 0.01). In contrast, no protection was observed with either the Bretschneider or Rose solutions: Cardiac output recovered to 31.7% +/- 10.3% and 5.1% +/- 3.2%, respectively, in these groups. Postischemic creatine kinase leakage was 72.4 +/- 12.3 and 92.1 +/- 18.6 IU/15 min/gm dry weight in the St. Thomas' Hospital and Tyers groups compared with 125.6 +/- 28.6 IU/15 min/gm dry weight in control hearts (p = no significant difference). In the Bretschneider group, creatine kinase leakage increased to 836.9 +/- 176.8 IU/15 min/gm dry weight (p less than 0.01 versus noncardioplegic control hearts), and with the Roe solution the value was 269.0 +/- 93.0 IU/15 min/gm dry weight (p = no significant difference). In conclusion, cardioplegic protection can be achieved in the immature rabbit myocardium with both St. Thomas' Hospital and Tyers solutions, but acalcemic solutions such as Bretschneider and Roe solutions (which may be effective in the adult heart) increased damage in this preparation. The reported lack of cardioplegic efficacy in the immature myocardium may therefore reflect the choice of cardioplegic solution rather than a greater vulnerability to injury in the neonatal heart.     

维生素E对膀胱出口部分梗阻后膀胱功能的保护作用

目的探讨维生素E对兔膀胱出口部分梗阻引起膀胱功能改变的保护作用.方法新西兰雄兔28只随机分为A组6只、B组6只、C组8只、D组8只,A、B、C组正常饮食,D组每日给予维生素E 600 mg,4周后B组建立假手术模型,C、D组建立膀胱出口部分梗阻模型.术后4周各组进行尿动力学检查、膀胱称重、RT-PCR检测膀胱组织肌质网钙泵蛋白(SERCA2)mRNA水平、Western blot检测SERCA2和肌动蛋白表达水平.结果正常A组和假手术B组各项参数比较差异均无统计学意义,合并为对照组(A+B组).膀胱重量C组为(13.07±1.71)g、D组为(11.80±2.01)g,约为对照组(2.81±0.30)g的4倍(P＜0.01).尿动力学检查最大逼尿肌压力D组为(37.38±4.04)cm H2O,大于C组的(24.13±4.54)cm H2O和对照组的(22.70±1.89)cm H2O(P＜0.05);膀胱容量D组为(83.00±13.05)ml、对照组为(67.00±7.22)ml,均大于C组的(45.13±6.63)ml(P＜0.05);膀胱顺应性D组为(8.18±1.95)ml/cm H2O、对照组为(6.67±0.90)ml/cmH2O,均好于C组(3.35±0.68)ml/cm H2O(P＜0.05);SERCA2 mRNA表达D组为1.45±0.16、对照组为1.41±0.05,高于C组的0.97±0.11(P＜0.05);SERCA2蛋白表达D组为1.90±0.19、对照组为2.18±0.23,高于C组的1.35±0.16(P＜0.05);而三组肌动蛋白表达差异无统计学意义.结论预先服用维生素E可以提高梗阻后SERCA2基因转录和表达水平,可能是保护膀胱功能的机制之一.     

Pharmacological protection of rabbit kidneys from normothermic ischemia.

This study investigated the influence of the administration of pharmacological agents on the recovery of rabbit kidneys from the effects of 1 hr of situ normothermic ischemia, utilizing acute and chronic models. The agents tested included the diuretics mannitol and furosemide, the vasoactive agents phenoxybenzamine, propranolol, and dopamine, and the membrane stabilizers chlorpromazine and methylprednisolone. A beneficial effect was detected only with the diuretic agents and propranolol when given prior to the ischemic insult.     

Resistance and tolerance of the myocardium to ischemia.

Since 1967, the authors have abandoned coronary perfusion in valve, particularly aortic, surgery. Some of the difficulties encountered during defibrillation at the period where coronary perfusion was always used have dramatically decreased. Extra-corporeal-circulation is now performed under moderate hypothermia--28 to 30 degrees C--which gives excellent myocardial protection for aortic cross clamping time of 30 to 60 minutes. In coronary artery surgery, the same technique is now used, after having wrongly believed previously that coronary perfusion was indicated. In conclusion, we feel that coronary perfusion is not indicated any more but in very exceptional cases.     

Efficacy and vasodilatory benefit of magnesium prophylaxis for protection against spinal cord ischemia

Prevention of paraplegia remains an imperative issue in thoracoabdominal aortic surgery. The aim of this study was to assess the efficacy of a prophylactic magnesium infusion in a rat spinal cord ischemia model and to demonstrate spinal blood flow increase caused by the infusion. The study was conducted in two parts. Firstly, the neuroprotective effect of magnesium was assessed using a rat model with two different ischemic times: 10 min and 14 min. Spinal cord ischemia was induced by occlusion of the descending aorta. Rats in the treatment group were given a 100 mg/kg magnesium sulfate infusion before ischemia. Secondly, relative changes in spinal cord blood flow before and during ischemia were recorded using the laser Doppler flowmetry technique. Changes in blood flow were compared between the magnesium and control groups. Rats pretreated with magnesium showed good overall recovery after both 10 min (incidence of paraplegia 62.5% control vs. 37.5% Mg, n = 8 each) and 14 min (85.7% control vs. 57.1% Mg, n = 7 each) of ischemia, although the differences compared with controls were statistically insignificant. However, the magnesium group showed significantly better neurological performance during the early postischemic period. Comparison of changes in spinal circulation revealed less reduction in blood flow during ischemia in the magnesium-treated group. In conclusion, magnesium may have potential prophylactic benefits during ischemia by exerting a neuroprotective effect through vasodilation of the spinal cord vasculature. To our knowledge, this vasodilatory effect on the spinal cord has not previously been investigated. Optimization of the treatment regimen, however, is required.     

梯度灌注对沙土鼠全脑缺血神经元凋亡的保护作用

目的　观察梯度灌注时间和灌注量对全脑组织缺血损害的恢复程度和凋亡基因的表达 ,探讨梯度灌注的脑保护作用。方法　将沙土鼠随机分 4组 ,实验组夹闭两侧颈总动脉 ,造成沙土鼠全脑缺血模型 ,夹闭 10min后 ,开放 10min ,开放不同脑血流量 (1/4、1/2、1/1)和单纯血液稀释后分别观察缺血海马CA区凋亡蛋白Bcl x/l和Bax的表达 ,海马区神经元的凋亡形态学改变和缺血区大脑半球的改善状况。结果　开放 10min ,1/2脑血流量时Bcl x/l蛋白表达较一次性再灌注明显上调 (P <0 .0 5 ) ,而Bax表达下调。海马CA区缺血损害改善最明显 ,开放 10min、全脑血流量一次性开放时海马CA区损害最严重。结论　梯度灌注流量有明显的抗脑缺血海马区神经元凋亡作用 ;低灌注流量有较好的改善脑缺血作用 ;夹闭 10min后 ,开放 1/2脑血流量 ,开放 10min的效果比一次性再灌注开放效果要好。     

Studies of myocardial protection in the immature heart. IV. Improved tolerance of immature myocardium to hypoxia and ischemia by intravenous metabolic support

Thirteen immature puppies (2 to 4 kg) underwent 1 hour of acute hypoxia (oxygen tension 25 to 30 mm Hg), followed by 45 minutes of normothermic global ischemia on total vented bypass with normal blood reperfusion. Ventricular function was assessed by inscribing Starling function curves and measuring stroke work indices before hypoxia and after reperfusion. Seven puppies (control) received normal saline infusion at 4 ml/kg/hr. Six other puppies received a 4 ml/kg/hr intravenous infusion of glutamate/aspartate, glucose-insulin-potassium, mercaptopropionyl glycine, carnitine, and catalase during hypoxia and reperfusion. In control hearts, acute hypoxia depleted myocardial glutamate and aspartate by 52% (p less than 0.05 versus prehypoxia) and 48% (p less than 0.05 versus prehypoxia) and caused severe hemodynamic deterioration (55% decrease of stroke work index) (p less than 0.05 versus prehypoxia); three of seven (43%) required premature institution of bypass. Postischemic left ventricular function recovered to only 40% of control levels (p less than 0.05 versus prehypoxia). In contrast, intravenous metabolic infusions maintained tissue glutamate (p less than 0.05 versus control group) and aspartate (p less than 0.05 versus control group) in treated hearts during hypoxia and allowed cardiac index to rise 20% (p less than 0.05 versus prehypoxia); all treated hearts tolerated 1 hour of hypoxia, and stroke work recovered 70% (p less than 0.05 versus control group) of stroke work index after subsequent ischemia. Impaired tolerance of immature hearts to acute hypoxia and subsequent ischemia is due to substrate depletion. This impairment can be reduced by intravenous metabolic support during hypoxia and reperfusion and leads to improved recovery of postischemic function.     

不同晶体停搏液对幼兔心肌作用的比较

目的比较四种停搏液[改良托马氏液1号-MST,台氏液（Tyers）,布氏液（Bretshneider—HTK）,罗氏液（Roe）]对未成熟心肌的作用。方法观察不同停搏液模型幼兔心在Langendorff离体灌注装置14℃缺血2h后血流动力学、冠脉流出液心肌酶和心肌生化的变化。结果与Krebs—Henseleit重碳酸盐缓冲（KH）液比较,MST和Tyers停搏液可明显提高幼兔心功能的恢复（P〈0．05）,心肌酶的漏出较少（P〈0．05）,ATP储存较好（P〈0．05）。与KH液比较,HTK和Roe停搏液模型幼兔心功能恢复差（P〈0．05）,心肌酶漏出明显增多（P〈0．05）。结论MST和Tyers停搏液对未成熟心肌可提供较满意的心肌保护,HTK和Roe停搏液不能对未成熟心肌提供良好的心肌保护。     

Metabolic and structural recovery of left ventricular canine myocardium from regional complete ischemia.

The capacity for recovery of the normothermic left ventricular myocardium from a regional complete ischemia (RCI) was investigated using changes in the myocardial metabolic status (ATP, ADP, AMP, creatine phosphate (CrP), free creatine, glycogen, glucose, lactate) and alterations of the morphology as parameters. In dogs, an area of the anterior wall of the left ventricular myocardium was temporarily deprived completely of its blood supply by 5--7 overlapping ligatures extending into the heart cavity. The metabolites of the adenylic acid-CrP system returned to normal tissue levels after 30 and 60 min of RCI within 14 and 35 days of recovery, respectively; restoration averaged 82% after 100 min, 74% after 140 min, and 38% after 180 min of RCI after 5 weeks of recovery. At the same time glycogen amounted to 163% after 100 min, 114% min, and 65% after 180 min of RCI. The biochemical data correlated well with the structural changes in the affected myocardium, especially with the amount of de- and regenerating heart muscle cells. These obviously were functionally defect and were not comparable with normal structured and functioning heart muscle cells.