Morphological characteristics of tissue components of different layers of the rat myocardium

The tissue components of the subendocardial, inner and outer intramural layers of the myocardium were examined by morphometry. There was no significant difference in the proportion of cardiomyocytes in the different layers of the myocardium (subendocardium 0.820 +/- 0.007; inner layer 0.713 +/- 0.100; outer intramural layers 0.727 +/- 0.008; subepicardium 0.699 +/- 0.009). The relative surface of cardiomyocytes was maximal in the subepicardium (58.62 +/- 1,18). The magnitudes of the volumetric density and surface of the capillaries decreased from the subepicardial toward the subendocardial layer. The diameter of myocytes in the test layers of the myocardium varied within a wide range.     

Formation of Cardioelectric Field on the Body Surface at a Period of Activation of Ventricular Myocardium in the Chicken Gallus domesticus

Spatial and temporal non-uniform and polyfocal depolarization of the subendocardial, intramural, and subepicardial layers of the ventricle myocardium in the chicken have been established experimentally. Different depth and time of formation of activation centers in the ventricular myocardium provide the appearance of groups of multiple depolarization foci on the epicardial surface of the ventricles. During the initial ventricular activity the cardioelectric field (CEF) on the chicken body surface is characterized by three periods of the dynamics of distribution of potentials: (1) the period of their gradual changes reflecting the electrical activity of excitation foci in the subendocardial, intramural, and subepicardial ventricular layers of myocardium on CEF; (2) the period of inversion consisting of an alteration of the mutual arrangement of the positive and negative CEF areas, this alteration corresponding in time to polyfocal depolarization of the epicardial surface of the ventricles; (3) the period of stability, during which the arrangement of the positive and negative CEF regions does not change, which is due to depolarization of multiple myocardium zones at the final phase of the heart ventricle activation.     

Stereological analysis of the parenchymal and stromal structures of a hypertrophied myocardium in acute arterial hypertension

Arterial hypertension in 35 male Wistar rats was produced by disturbance of the left renal artery circulation. Myocardial tissue reorganization was studied by using the methods of light microscopy and stereological analysis. By the 35th day of the experiment marked alterations of the intramural vessels were found which were manifested in the thickening of the vessel walls due to hyperplasia and hypertrophy of the smooth muscles cells, and in the developing of sclerotic processes in all layers of the arterial walls. At the tissue level a decrease of the volume and surface densities of capillaries and connective tissue cells were determined, that resulted in a decline of the ratio between the volume and surface densities of the structures to the volume density of cardiomyocytes. Informational analysis revealed an increase of entropy and relative entropy of the myocardium tissue during its hypertrophy.     

Ischemia-induced changes in sarcolemmal (Na+, K+)-ATPase, K+-pNPPase, sialic acid, and phospholipid in the dog and effects of the nisoldipine and chlorpromazine treatment

This work was undertaken to study functional and structural changes of the cardiac sarcolemmal membrane which was isolated from the ischemic lesion in the dog. The sarcolemmal fraction was prepared, by adopting the method devised by Reeves and Sutko , from the right ventricle and the subendocardial and subepicardial layers of the left ventricle. Ischemic lesion was produced by occlusion of a branch of the left anterior descending coronary artery for a period of 1.5 hr in the thoracotomized dog, followed by release of the occlusion for 3 hr. Nisoldipine, 5 micrograms/kg, was given twice intravenously, and chlorpromazine was infused at a rate of 10 micrograms/kg X min, in addition to the administration of twice bolus doses of 400 micrograms/kg each. Nisoldipine significantly decreased the incidence of premature ventricular contractions and microvascular hemorrhage. Sarcolemmal purity was monitored by using enzyme and chemical markers; the results indicated that the membrane preparation was tenfold purified over the homogenate. Although the activities of ouabain-sensitive (Na+, K+)-ATPase and ouabain-sensitive K+-p-nitrophenylphosphatase ( pNPPase ) of the sarcolemmal preparation isolated from the subendocardial layer were similar to those from the subepicardial layer in the nonischemic left ventricle, a significant decrease in these activities was observed only when the sarcolemmal fraction isolated from the subendocardial layer of ischemic area was compared with that from the subendocardial layer of nonischemic area. In contrast, the sialic acid content of the sarcolemma from the ischemic subendocardial layer was significantly increased compared to that of the nonischemic subendocardial layer. No such changes occurred in sarcolemma prepared from the ischemic subepicardial layer. The total phospholipid content as well as phosphatidylcholine and -ethanolamine contents of the sarcolemmal membrane prepared from the subendocardial layer of ischemic area were significantly decreased compared to nonischemic area. Nisoldipine prevented the ischemia-induced alterations in sarcolemmal (Na+, K+)-ATPase, pNPPase , sialic acid and phospholipids of the subendocardial layer. Chlorpromazine showed a less consistent effect than did Nisoldipine under our experimental conditions. Our study thus demonstrates that the lipid component and function of cardiac sarcolemmal membrane are altered in the early ischemic lesion and that these alterations are nonuniform in distribution and are alleviated by some pharmacological intervention.     

心室不同部位起搏对正常犬和扩张型心肌病心力衰竭犬模型在体心肌跨室壁复极离散的影响

实验以正常犬和扩张型心肌病心力衰竭犬(dilated cardiomyopathy congestive heart failure,DCM-CHF)模型为对象、以心肌跨室壁复极离散的相关参数为指标,研究左心室心外膜起搏、双心室起搏(模拟临床上心室再同步治疗的方法)后的心肌电生理特性变化。实验以快速右心室起搏的方法制备DCM-CHF犬模型;正常犬和DCM-CHF犬均经射频消融希氏束制备三度房室传导阻滞模型;采用同步记录犬体表心电图和内膜下、中层、外膜下三层心肌单相动作电位(monophasic action potentials,MAP)的方法,测定不同部位起搏时的QT间期、Tpeak-Tend(Tp-Te)间期和三层心肌的单相动作电位时程(MAP duration,MAPD)、跨室壁复极离散度(transmural dispersion of repolaization,TDR)。结果显示:在正常犬,左室心外膜与双心室起搏后三层心肌的MAPD均延长,同时TDR增大(左室心外膜起搏47.16 ms、双心室起搏37.54 ms、右室心内膜起搏26.75 ms,P<0.001),体表心电图Tp-Te间期的变化与之平行;在DCM-CHF犬较正常犬已表现出中层心肌MAPD延长(276.30 ms vs 257.35 ms,P<0.0001)和TDR(33.8 ms vs 27.58 ms,P=0.002)增大的基础上,左室心外膜参与起搏后仍进一步使三层心肌的MAPD延长和TDR增大。研究结果提示,左室心外膜起搏和双心室起搏后使内膜下、中层     

Subendocardial and subepicardial pressure-flow relations in the rat heart in diastolic and systolic arrest

Ischemic heart disease is more apparent in the subendocardial than in subepicardial layers. We investigated coronary pressure-flow relations in layers of the isolated rat left ventricle, using 15 microm microspheres during diastolic and systolic arrest in the vasodilated coronary circulation. A special cannula allowed for selective determination of left main stem pressure-flow relations. Arterio-venous shunt flow was derived from microspheres in the venous effluent. We quantitatively investigated the pressure-flow relations in diastolic arrest (n=8), systolic arrest at normal contractility (n=8) and low contractility (n=6). In all three groups normal and large ventricular volume was studied. In diastolic arrest, at a perfusion pressure of 90 mmHg, subendocardial flow is larger than subepicardial flow, i.e., the endo/epi ratio is approximately 1.2. In systolic arrest the endo/epi ratio is approximately 0.3, and subendocardial flow and subepicardial flow are approximately 12% and approximately 55% of their values during diastolic arrest. The endo/epi ratio in diastolic arrest decreases with increasing perfusion pressure, while in systole the ratio increases. The slope of the pressure-flow relations, i.e., inverse of resistance, changes by a factor of approximately 5.3 in the subendocardium and by a factor approximately 2.2 in the subepicardium from diastole to systole. Lowering contractility affects subendocardial flow more than subepicardial flow, but both contractility and ventricular volume changes have only a limited effect on both subendocardial and subepicardial flow. The resistance (inverse of slope) of the total left main stem pressure-flow relation changes by a factor of approximately 3.4 from diastolic to systolic arrest. The zero-flow pressure increases from diastole to systole. Thus, coronary perfusion flow in diastolic arrest is larger than systolic arrest, with the largest difference in the subendocardium, as a result of layer dependent increases in vascular resistance and intercept pressure. Shunt flow is larger in diastolic than in systolic arrest, and increases with perfusion pressure. We conclude that changes in contractility and ventricular volume have a smaller effect on pressure-flow relations than diastolic-systolic differences. A synthesis of models accounting for the effect of cardiac contraction on perfusion is suggested.     

Transmural non-homogeneity of calcium-induced heart injury

The Ca paradox resulted in marked inhibition, up to disappearance, of histochemically studied enzyme activities (SDH, LDH, beta-HBDH, phosphorylase and ATPase) in the subepicardial layer of the myocardium. In the subendocardial region there was only a small decrease. These transmural differences correlated well with ultrastructural changes. It is assumed that the heterogeneity in transmural distribution of injury is the result of transmural differences in coronary flow.     

Sequence of depolarization of atrial myocardium of the pig <Emphasis Type="Italic">Sus scrofa domesticus</Emphasis>

By the method of multichannel synchronous cardioelectrotopography, sequence of depolarization of intramural atrial layers was studied in representative of ungulate animals-pigs. In the pig atrial myocardium there is revealed a complex picture of spreading of activation fronts, connected with non-uniform anatomical structure and subendocardial (intramural) disposition of area of initial activation. The general character of sequence of depolarization of the pig atrial myocardium is comparable with carnivores (dog) and even-toed (ship) animals.     

Proliferative processes in the myocardium in different parts of the heart during the creation of experimental myocardial infarct of the left ventricle in 2- and 3-week-old rats

As a result of 30 times repeated injections of 3H-thymidine (3HTdr) to neonate rats, beginning from days 13 or 21 post partum, ca. 20 and 10% of myonuclei in the left and right atria were labeled, respectively, while in both ventricles cumulative labeling of myocytes was nearly ten times lower. In rats of the same age with experimental infarction of the left ventricular myocardium the number of myonuclei labeled after 30-fold 3HTdr injections increased in atria up to 40-50%, in perinecrotic myofibers of the left ventricles up to 8-11%, and in myofibers of the left and right ventricle located far from the necrotic foci up to 3-4 and 2-3%, respectively. In some of rats subendocardial and/or subepicardial layers of the surviving left ventricular myocardium contained up to 15-35% of labeled myonuclei. Thus, in neonatal rats the extent of DNA synthesis reactivation in the nuclei of cardiomyocytes, the majority of which have recently completed normal ontogenetic proliferation, is, on the whole, of the same order as found in similar experiments on adult rats (Rumiantsev, Kassem, 1976; Oberpriller et al., 1984). However, still immature ventricular myocytes of neonatal rats resume mitotic cycle easier than those of adult animals which is evidenced not only by higher numbers of 3HTdr labeled myonuclei in subepicardial and subendocardial ventricular myocardia of some rats, but even more by reactivation of DNA synthesis in a limited fraction (2-3%) of the whole population of non-perinecrotic myocytes in both ventricles. Besides, reactive proliferation of cardiomyocytes in the atria of neonate rats, unlike in adults, starts on day 3 rather than on day 5 after infarction is induced. In the atria of neonatal rats polyploidization of myonuclei at later postinfarction stages is less pronounced than in adult rats which may be accounted for by formation of individual daughter nuclei during acytokinetic mitoses or, more seldom, by completion of cytotomy.     

Cardiac hypertrophy in rats after supravalvular aortic constriction. I. Size and number of cardiomyocytes, endothelial and interstitial cells

The ascending aorta of 22 adult male Sprague-Dawley rats was constricted with a silver ring, and 25 animals were subjected to a sham-operation. The hearts, including the main arteries, were fixed by retrograde perfusion 3, 7, 14, 21 and 35 days after the operation. The cross-sectional area of the aorta was reduced by the constriction to an average of 20% of the values found after sham-operation. Twenty-one days after the constriction the weight of the left ventricular myocardium including the septum was increased 1.7-fold compared with controls. No further increase in weight was observed 35 days after the operation. The relative volumes of the tissue components remained largely constant in the subepicardial myocardium. In the subendocardial myocardium, however, the volume fraction of interstitial and, to a lesser extent, of endothelial tissue was significantly increased. Twenty-one days after constriction the estimated total volumes of the different myocardial components per left ventricle were increased 1.7-fold for heart muscle parenchyma, 1.8-fold for endothelial tissue, 2.9-fold for interstitial tissue, and 1.3-fold for capillary lumina compared with controls. At 35 days, only the interstitial tissue showed a further increase to 4.8-fold of control values. The mean cardiomyocyte volume was increased after aortic constriction in proportion to the increase in left ventricular weight, i.e. 1.7-fold over controls at 21 days. After 35 days its value was 29,500 +/- 790 micron 3 in rats subjected to aortic constriction compared with 16,800 +/- 640 micron 3 in controls. At this time the estimated number of cardiomyocytes per left ventricle showed no significant differences between experimental animals (2.9 X 10(7)) and controls (3.1 X 10(7)). Endothelial and interstitial cells were not only increased in average single cell volume (1.3-fold and 2.0-fold, respectively), but also in number per left ventricle (1.4-fold and 2.7-fold, respectively). Two-dimensional parameters indicated that during hypertrophy the capillary supply lagged behind the overall mass increase but achieved control levels on termination of hypertrophic growth at 35 days. These results show that even in pronounced hypertrophy the increase in mass of the myocardial parenchyma in the rat is due exclusively to an enlargement of cardiomyocytes (hypertrophy), whereas in endothelial and interstitial tissues enlargement of cells as well as increase in cell number (hyperplasia) also plays a role.     

Intramyocardial pressure and coronary extravascular resistance

Intramyocardial pressure is an indicator of coronary extravascular resistance. During systole, pressure in the subendocardium exceeds left ventricular intracavitary pressure; whereas pressure in the subepicardium is lower than left ventricular intracavitary pressure. Conversely, during diastole, subepicardial pressure exceeds both subendocardial pressure and left ventricular pressure. These observations suggest that coronary flow during systole is possible only in the subepicardial layers. During diastolic, however, a greater driving pressure is available for perfusion of the subendocardial layers relative to the subepicardial layers. On this basis, measurements of intramyocardial pressure contribute to an understanding of the mechanisms of regulation of the phasic and transmural distribution of coronary blow flow.     

Effect of localization of the ectopic excitation site on the sequence and duration of depolarization in the ventricular epicardium in dogs

The depth of the myocardial wall ectopic focus was found to affect spatial and temporal characteristics of the depolarization process in the heart ventricular surface. Duration of the ventricular epicardial depolarization under the ectopic foci located in subendocardial and intramural layers of the myocardium was shorter than in epicardial stimulation of the ventricles. A dependence of the ectopic excitation duration on the pacing site localization in the epicardium, was revealed. The shortest duration of the depolarization occurred under electrical stimulation of the apex and ventral part of the interventricular septum, whereas the longer one--under pacing the left ventricular base.     

Identification and mapping of brain natriuretic peptide in the normal ventricular myocardium of a desert-dwelling mammalian model,the camel (Camelus dromedarius): Immunohistochemical and ultrastructural study

Brain natriuretic peptide (BNP) is mainly produced in the ventricular myocardium, where it is released into the circulation, producing rapid volume decrease by diuresis, natriuresis, and water shift into the extracellular space, and vasodilation. The dromedary camel, a mammalian model of the desert nomads, lives under unfavorable physiological stresses during thirst, starvation, desiccation, and hot climate, thus has a special demand for water homeostasis. The present studies characterized BNP in the ventricular myocardium of healthy camels, immunohistochemically with a specific antibody, and ultrastructurally identified the endocrine property of the cardiomyocytes and Purkinje fibers. The paranuclear, granular, immunoreactive material was not restricted to the cardiomyocytes, as it was also visible in the Purkinje fibers and their associated nerve varicosities. The intensity of immunoreactive BNP showed a transmural gradient from the subepicardium to the myocardium. Intense immunoreactivity was also noted among the perivascular cardiomyocytes. At the electron microscopic level, specific granules were demonstrated in the paranuclear cytosol of cardiomyocytes and Purkinje fibers. The current study provides the first immunohistochemical localization pattern of BNP in the camel myocardium and suggests a relationship between the intense subepicardial BNP-immunoexpression and a possible translocation of the active hormone to the pericardial fluid for further paracrine actions on the heart and its coronaries.     

Histostereologic analysis of the myocardium of homoiothermic animals during cooling

A stereologic study of the myocardium exposure to cold for 4 hours, 8 and 16 days was carried out on Wistar rats. It has been shown that during the first 8 days acute hemodynamic disorders and cardiomyocyte contractures developed. These changes were followed by a decrease in the volume and surface capillary density resulting in the impairment of transcapillary exchange and reduction of the intracellular regeneration processes in cardiomyocytes. By the 16th day the compensatory-adaptive reactions developed, i.e., the volume and surface density of endothelial cells and capillaries was increased. At the same time the lysis processes caused by the general decrease in the inflow of plastic substances to the myocardium due to the increase in thermogenesis were revealed in part of cardiomyocytes. These changes in parenchymatous cells were accompanied by the intensification of desmoplastic reactions in the myocardial stroma.     

Effects of red blood cell aggregation on myocardial hematocrit gradient using two approaches to increase aggregation

The normal transmyocardial tissue hematocrit distribution (i.e., subepicardial greater than subendocardial) is known to be affected by red blood cell (RBC) aggregation. Prior studies employing the use of infused large macromolecules to increase erythrocyte aggregation are complicated by both increased plasma viscosity and dilution of plasma. Using a new technique to specifically alter the aggregation behavior by covalent attachment of Pluronic F-98 to the surface of the RBC, we have determined the effects of only enhanced aggregation (i.e., Pluronic F-98-coated RBCs) versus enhanced aggregation with increased plasma viscosity (i.e., an addition of 500 kDa dextran) on myocardial tissue hematocrit in rapidly frozen guinea pig hearts. Although both approaches equally increased aggregation, tissue hematocrit profiles differed markedly: 1) when Pluronic F-98-coated cells were used, the normal transmyocardial gradient was abolished, and 2) when dextran was added, the hematocrit remained at subepicardial levels for about one-half the thickness of the myocardium and then rapidly decreased to the control level in the subendocardial layer. Our results indicate that myocardial hematocrit profiles are sensitive to both RBC aggregation and to changes of plasma viscosity associated with increased RBC aggregation. Furthermore, they suggest the need for additional studies to explore the mechanisms affecting RBC distribution in three-dimensional vascular beds.     

Regional metabolic rate of exogenous glucose in the isoprenaline and dobutamine stimulated canine myocardium as estimated by the 2-deoxy-d[1-14C]glucose method

The effect of β-adrenoceptor stimulation by isoprenaline and dobutamine on the transmural distribution pattern of regional myocardial metabolic rate of exogenous glucose (RMMRGlc) was studied in the anesthetized closed chest dog using the 2-deoxy-d[1-¹⁴C]glucose method. In a previous series a lumped constant (LC) value of 0.93 ± 0.47 (1 SD) was measured for [¹⁴C]2-deoxyglucose in the canine myocardium. In the control group (N = 12) RMMRGlc was significantly higher in the subendocardial layer of the left ventricular free wall than in both the middle and subepicardial layer, where it was quite evenly distributed (P ⩽ 0.05). With i.v. dobutamine (N = 8) RMMRGlc was significantly lower in the midportion of left ventricular free wall than in the subepicardial layer (P ⩽ 0.05), but it was not different from the inner wall section. Significant differences between the subepicardial and subendocardial portions of the left ventricular free wall could not be found, either. In the isoprenaline group (N = 9) no transmural gradients of RMMRGlc were observed in the left ventricular myocardium. In all groups, both the interventricular septum and the right ventricular free wall exhibited homogeneous distribution patterns of RMMRGlc.It is concluded that transmural distribution patterns of exogenous glucose utilization probably reflect corresponding gradients in energy demands of the left ventricular wall. Redistribution of RMMRGlc in the isoprenaline and dobutamine groups may result from altered working conditions, a change in local inotropic state of the left ventricular myocardium, or from regional differences in the proportions of substrate utilization, and from regional differences in adrenoceptor density.     

兔心室肌细胞电生理异质性研究--缺血对动作电位和钾流的影响

实验用全细胞膜片箝技术,观察正常及缺血条件下,兔心内膜下心室肌细胞与心外膜下心室肌细胞的动作电位和稳态外向钾流及其变化。结果显示：（１）正常条件下,心外膜下心室肌细胞与心内膜下心室肌细胞动作电位形态有差异,心外膜下心室肌细胞动作电位时程（ＡＰＤ）较短,复极１期后有明显的初迹,动作电位形态是“锋和圆顶”,而心内膜下心室肌细胞ＡＰＤ较长,并且没有上述动作电位形态特征。这两类细胞静息电位无差异。（２）在     

压力负荷性心衰小鼠左心室跨壁L-型钙电流的变化

为了观察正常和心衰时心内膜下和心外膜下心肌细胞L-型钙电流(ICa-L)的差别,我们采用主动脉弓狭窄的方法建立小鼠压力超负荷性心衰模型,采用全细胞膜片钳技术记录了正常、主动脉狭窄(band)及假手术对照(sham)组动物左心室游离壁内、外膜下心肌细胞的动作电位时程(action potential duration,APD)和ICa-L。结果显示:(1)与sham组同龄的正常小鼠左心室心内膜下细胞动作电位复极达90％的时程(APD90)为(38．2±6．44)ms,较心外膜下细胞的APD90(15.67±5.31)ms明显延长,二者的比值约为2．5:1;内膜下细胞和外膜下细胞ICa-L密度没有差异,峰电流密度分别为(-2.7±0.49)pA/pF和(-2.54±0．53)pA/pF;(2)Band组内、外膜下细胞的动作电位复极达50％的时程(APD50)、APD90均较sham组显著延长,尤以内膜下细胞延长突出,分别较sham组延长了400％和360％,内、外膜下细胞APD90的比值约为4.2:1;(3)与sham组相比, band组内膜下细胞ICa-L密度显著减小,在+10 mV～+40 mV的4个电压下分别降低了20．2％、21．4％、21.6％和25.7％(P< 0．01),但其激活电位、峰电位和翻转电位没有改变;band组外膜下细胞的ICa-L密度与同期sham组相比无明显变化;band组钙通道激活、失活及复活的动力学特征与sham组相比没有改变。以上结果提示,生理状态下小鼠左心室内、外膜下细胞ICa-L密度不存在明显差别,提示ICa-L与APD跨壁异质性的产生无关;心衰时左心室内、外膜下细胞APD明显延长,以内膜下细胞延长尤为突出,内膜下细胞ICa-L密度明显减少,而外膜下细胞ICa-L密度无明显改变,这种ICa-L的非同步变化在心衰时可能起到对抗APD延长、减少复极离散度的有益作用。     

Myocardial blood flow regulation relative to left ventricle pressure and volume in anesthetized dogs

The influence of left ventricle pressure and volume changes on coronary blood flow was investigated in eight anesthetized dogs. Coronary artery pressure-flow relationships were determined at two levels of left ventricular pressure and volume. The distribution of blood flow within the myocardium was also determined when these relationships varied. Reducing left ventricle pressures and volumes increased heart rate. Rate-pressure product, diastolic coronary pressure, myocardial O2 consumption, total, subendocardial and subepicardial flow decreased. Hematocrit and blood gas data were unchanged. The pressure-flow relationships were shifted leftward (p = 0.001) but the range of autoregulation was not altered. At low left ventricle pressures and volumes, the lower coronary artery pressure limit was shifted leftward (from 75 to 45 mm Hg (1 mm Hg = 133.3 Pa)), while total, subendocardial, and subepicardial blood flow did not change compared with the control. Below the lower coronary artery pressure limit, subendocardial but not subepicardial flow decreased, resulting in maldistribution of flow across the left ventricular wall. When coronary pressure was reset between control and the lower coronary artery pressure limit, subendocardial flow was restored. These results show that the lower coronary artery pressure limit can be shifted leftward while the distribution of blood flow across the left ventricular wall is preserved.