Izuru Nakamura

Mitochondrial DNA deletion in human myocardium

Mutation of myocardial mitochondrial DNA was investigated in human left ventricles obtained at autopsy using the polymerase chain reaction (PCR). Seventeen autopsy cases were examined, including patients with diabetes mellitus, myocardial infarction, cardiomyopathy, cancer, and other diseases. Two cases of diabetes mellitus, 2 of myocardial infarction, and 1 of pulmonary fibrosis showed a 7.4 kb deletion of myocardial mitochondrial DNA. Primer shift PCR confirmed that an amplified DNA fragment had not been obtained by misannealing of the primers. It is unclear how much these findings are related to the severity or prognosis of the various diseases, but they indicate that mutation of myocardial mitochondrial DNA can occur in other diseases besides cardiomyopathy, although the influence of aging could not be excluded.

Mechanical catecholamine responsiveness and myosin isoenzyme pattern of pressure-overloaded rat ventricular myocardium

SummaryPressure-overloaded cardiac hypertrophy was induced by abdominal aortic constriction in 10-week-old male Wistar rats. 24–26 weeks after aortic constriction, the hearts were excised and a myocardial mechanical study was performed using isolated left ventricular papillary muscles. There was no significant difference in isometric developed tension (T) between sham-operated control and aortic constriction (AC) rats (control vs AC rats=2.9±0.6 vs 2.7±0.7 g/mm2). dT/dtmax of AC rats, on the other hand, was significantly lower than that of controls (controls vs AC rats=32.8±7.5 vs 26.3±6.1 g/mm2sp<0.05). Myocardial mechanical responses to isoproterenol (10−7 mol/l) were depressed in the group with aortic constriction compared with the control group (ΔT:18.5±6.7 vs 12.1±4.9%,p<0.05, ΔdT/dt: 25.2±6.2 vs 17.5±5.8%,p<0.02). Responses of the parameters to dibutyryl cyclic AMP (10−5 mol/l) were also smaller in the AC group than in the control group (ΔT: 18.0±5.6 vs 13.3±4.0%,p<0.05, ΔdT/dt: 20.4±6.9 vs 14.7±4.1%,p<0.05). Left ventricular myosin isoenzyme pattern, revealed by pyrophosphate gel electrophoresis, shifted towards VM-3 under pressure overload. The present study demonstrates that post-membrane processes may be mainly responsible for the decreased myocardial mechanical catecholamine responsiveness in pressure-overloaded cardiac hypertrophy.

Myocardial Adaptive Changes and Damage in Ischemic Heart Disease

Changes in two of the elements of myocardial subcellular organelles relating to cardiac energetics, ventricular myosin isozymes and mitochondrial DNA mutations, were examined using left ventricular tissue samples obtained at autopsy from patients with ischemic heart disease. Myosin isozymes were examined in tissues from nine patients with ischemic heart disease and 12 control patients with cancer but no heart disease. Extracted myosin was separated by pyrophosphate gel electrophoresis. The relative concentration of each component was determined by densitometry. Mitochondrial DNA mutations were evaluated in tissues from ten patients with myocardial infarction and 11 control patients with cancer but no heart disease. DNA was extracted and mitochondrial DNA mutations were detected by the polymerase chain reaction. Two bands were revealed by pyrophosphate gel electrophoresis. These contained VM-A, which exhibited faster electrophoretic mobility and was present in lower concentrations, and VM-B, which had a lower mobility and a higher concentration, respectively. SDS polyacrylamide gel electrophoresis showed that these two components contained the heavy chain and light chains 1 and 2 of myosin. VM-A concentrations tended to be higher in patients with ischemic heart disease than in controls. A 7.4-kb deletion was detected between the D-loop and the ATPase 6 genes of mitochondrial DNA from the myocardium of 6 out of 10 patients with myocardial infarction. The relative amounts of the two myosin isozymes could be altered by ischemic heart disease, although the functional significance of these components is unclear. The changes in the two myosin isozymes might be an adaptive change to disordered energy metabolism, but this change was small. The myocardial mitochondrial DNA deletions in patients with myocardial infarction were thought to result from ischemic damage.

Effects of physical training on the myocardium of streptozotocin-induced diabetic rats

SummaryEffects of endurance swimming training on myocardial contractility and left ventricular myosin isoenzymes were examined in diabetic rats. A diabetic condition was induced in 15-weck-old male Wistar rats, by intravenous injection of streptozotocin (50 mg/kg). Swimming training was carried out for five to six weeks (90 min/day, 6 days/week). In order to estimate myocardial contractility, the isometric developed tension of the isolated left ventricular papillary muscle was measured. Myosin isoenzymes were obtained by pyrophosphate gel electrophoresis. Fasting blood glucose of the trained group was significantly lower than that of the sedentary group (sedentary vs. trained=409.6±25.9 vs. 266.3±20.5 mg/dl, p<0.001). There was no significant difference in isometric developed tension (T) between the two groups, and the dT/dtmax of the trained group showed a tendency to increase (sedentary vs. trained, T: 2.8±0.8 vs. 2.9±0.8 g/mm2, dT/dtmax: 23.1±3.6 vs. 26.2±3.5 g/mm2 · 2, p<0.1). Myocardial mechanical responses to isoproterenol and dibutyryl cAMP were increased in the trained group. Left ventricular myosin isoenzyme pattern was shifted towards VM-1 by endurance swimming (sedentary vs. trained, VM-1: 5.6±4.5 vs. 19.6±8.8%, p<0.001, VM-3: 75.1±10.0 vs. 54.9±14.7%, p<0.001). These results indicate that endurance swimming can improve disordered glucose metabolism and also influence myocardial contractility, myocardial catecholamine responsiveness, and energetics in myocardial contraction.

Myocardial contractility and left ventricular myosin isoenzyme pattern in cardiac hypertrophy due to chronic volume overload

Chronic volume overloaded cardiac hypertrophy was induced by abdominal arteriovenous shunt in 10-week-old male Wistar rats. Ten weeks after the operation, myocardial mechanical examination was performed with isolated left ventricular papillary muscles. Left ventricular myosin isoenzyme pattern was also examined by pyrophosphate gel electrophoresis. In addition, morphological study by electron microscope was performed. In arteriovenous shunt rats, isometric developed tension (T) and its first derivative (dT/dt) were decreased, resting tension (RT) was increased and time to peak tension (TPT) was prolonged as compared to sham-operated control rats (T: 2.4 +/- 0.5 vs 2.8 +/- 0.8 g/mm2, NS. dT/dt: 23.9 +/- 4.8 vs 31.9 +/- 7.0 g/mm2.s, p less than 0.05. RT: 1.2 +/- 0.2 vs 0.9 +/- 0.1 g/mm2, p less than 0.05. TPT: 151.7 +/- 20.2 vs 128.3 +/- 10.3 msec, p less than 0.05). Myocardial mechanical responses to isoproterenol (10(-7) mol/l) and dibutyryl cyclic AMP (10(-5) mol/l) were both depressed in hypertrophied myocardium, although not significant statistically. Left ventricular myosin isoenzyme pattern was shifted towards VM-3 by chronic volume overload. Electron microscope study revealed increased collagen content in hypertrophied myocardium.

Effects of long-term medication for essential hypertension on cardiac hypertrophy and function

The effects of long-term treatment of hypertensive patients with alacepril (angiotensin-converting enzyme inhibitor) on cardiac mass and function were investigated. A total of 12 patients was examined. Both systolic and diastolic blood pressure were significantly reduced by treatment with alacepril for 1 year. Left ventricular mass, as estimated by echocardiography, was significantly decreased by alacepril treatment, although electrocardiographic and chest x-ray findings were not significantly altered. Cardiac pump function, which was also assessed by echocardiography, was not changed. These results indicate that long-term treatment of hypertension with alacepril induces regression of cardiac hypertrophy without any change in cardiac contractile function.

Myocardial contractility and energetics in cardiac hypertrophy and its regression

The changes in myocardial contractility and ventricular myosin isoenzymes were investigated in rats with pressure-overload cardiac hypertrophy as well as during its regression. Hypertrophic myocardium was obtained from rats with renovascular hypertension (Goldblatt rats), rats with abdominal aortic constriction (AC), and spontaneously hypertensive rats (SHR). Regression of cardiac hypertrophy was induced by lowering the blood pressure through nephrectomy on the affected side in Goldblatt rats, by opening the clip which constricted the abdominal aorta in AC rats, and by the administration of antihypertensive agents to SHR. The isometric developed tension of isolated left ventricular papillary muscles and the maximum rate of increase in the tension (dT/dtmax) were measured. Left ventricular myosin isoenzymes were separated by pyrophosphate gel electrophoresis. Isometric developed tension remained unchanged, but dT/dtmax was decreased in hypertrophic myocardium, although it recovered along with the regression of cardiac hypertrophy. The left ventricular myosin isoenzyme pattern was shifted towards V3 in hypertrophic myocardium, and shifted back again towards V1 with the regression of cardiac hypertrophy. These results indicate that relief of hemodynamic overload is one of the most important elements in the regression of cardiac hypertrophy and the associated physiological or biochemical alterations. However, other factors such as neurohumoral influences must also be taken into consideration.

The Influence of Diabetes on Myocardial Contractility and Energetics in Spontaneously Hypertensive Rats

Diabetic patients with hypertension show a high incidence of the development of heart failure. The mechanisms involved in heart failure at the myocardial cellular level remain unclear. To elucidate the myocardial alterations occurring in hypertension associated with diabetes, we examined myocardial contractility and ventricular myosin isoenzymes in diabetic spontaneously hypertensive rats (SHR). Myocardial contractility was assessed by measuring the isometric tension developed in isolated left ventricular papillary muscles, and ventricular myosin isoenzymes were separated by pyrophosphate gel electrophoresis.