Biomedical subjects

K Shigenobu

Publications and source records attributed to K Shigenobu.

At least 109 records · Page 6Linked to original sources

PubMed1995

Cardiovascular selectivity of 1,4-dihydropyridine derivatives, efonidipine (NZ-105), nicardipine and structure related compounds in isolated guinea-pig tissues.

1. The cardiovascular selectivities of 1,4-dihydropyridine derivatives, efonidipine (NZ-105), nicardipine, 3NZ5NIC (the drug with NZ-105-type side-chain at C3 position and nicardipine-type at C5) and 3NIC5NZ (the drug with nicardipine-type side chain at C3 and NZ-105-type at C5) were studied in vitro. 2. All four compounds caused relaxation of guinea-pig aortae precontracted with a high K+. The pEC50 values were 7.5, 8.3, 8.1 and 5.6, for NZ-105, nicardipine, 3NIC5NZ and 3NZ5NIC, respectively. The relaxation produced by NZ-105 was slower in onset than those produced by the other compounds. The rate constant K(hr-1) of the relaxations were 0.59, 1.31, 1.02 and 1.24, for NZ-105, nicardipine, 3NIC5NZ and 3NZ5NIC, respectively. 3. In the electrically paced guinea-pig papillary muscles, NZ-105, 3NIC5NZ and 3NZ5NIC, even at concentrations as high as 10(-6) M, slightly decreased the contractile force (by 44.9 +/- 7.1%, 58.6 +/- 5.4% and 52.2 +/- 3.9%, respectively), whereas 10(-6) M nicardipine decreased the force by 84.9 +/- 3.3%. The negative inotropic effect of NZ-105 and 3NIC5NZ, but not that of 3NZ5NIC or nicardipine, was over 10 times weaker than their vasorelaxant effect. 4. In the guinea-pig right atria, NZ-105 and nicardipine at 10(-8) M decreased the spontaneous contraction rate by 67.9 +/- 15.0% and 39.7 +/- 15.4%, respectively. 3NIC5NZ at 3 x 10(-9) M and 3NZ5NIC at 3 x 10(-8) M had little effect on the rate, whereas 10(-8) M 3NIC5NZ and 10(-7) M 3NZ5NIC arrested the beating within 3 hr after administration.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1995

K+ channel-opening properties of a novel compound, NIP-121, in guinea pig myocardium as compared with those of cromakalim.

Myocardial effects of NIP-121, a novel compound with potent vasorelaxant activity, were examined in comparison with those of cromakalim in isolated tissue and cells from guinea pig hearts. NIP-121 and cromakalim concentration-dependently reduced the action potential duration (APD) of isolated papillary muscle; the effect was antagonized by glibenclamide. In isolated ventricular tissue, NIP-121 and cromakalim decreased the contractile force concentration dependently. In these two experiments, the potency of NIP-121 was approximately 20 times higher than that of cromakalim. The effects of NIP-121 and cromakalim on membrane currents were examined in voltage-clamped ventricular myocytes. NIP-121 (10(-6) M) and cromakalim (3 x 10(-5) M) increased the steady-state outward currents. The normal inwardly rectifying current-voltage relationship changed to a linear relationship that reversed at the K+ current reversal potential. The current activated by NIP-121 and cromakalim was inhibited either by glibenclamide or by increased intracellular ATP concentration. NIP-121, at this concentration, had little effect on the calcium current. Thus, NIP-121 was demonstrated to produce AP shortening and decrease in contractile force through activation of ATP-sensitive K+ currents in cardiac muscle, with a potency approximately 20 times higher than that of cromakalim.

Action Potentials↗

PubMed1995

Sustained negative inotropism mediated by alpha-adrenoceptors in adult mouse myocardia: developmental conversion from positive response in the neonate.

1. Inotropic responses to alpha-adrenoceptor stimulation and the effects of antagonists were examined in isolated ventricular preparations from neonatal and adult mice. 2. Phenylephrine, in the presence of propranolol, produced positive inotropic responses in neonates up to 1 week after birth, while it produced negative inotropic responses in mice older than 3 weeks. 3. Both positive and negative responses to phenylephrine in neonates and adults, respectively, were antagonized by prazosin, WB4101 (2-([2,6-dimethoxyphenoxyethyl]aminomethyl)-1,4-benzodioxane) and 5-methylurapidil, but not by atropine, yohimbine or chlorethylclonidine. 4. Noradrenaline (NA) produced positive inotropic responses both in the neonate and adult; the responses were observed in a lower concentration-range in the neonate than in the adult. WB4101 produced a significant leftward shift of the concentration-response curve for noradrenaline in adult preparations while only a slight rightward shift was observed in the neonate. 5. Our results demonstrate the presence of alpha-adrenoceptor-mediated inotropic responses in the mouse ventricular myocardia. The response to phenylephrine changes from a positive to a negative effect during postnatal development. The responses are mediated by alpha 1-adrenoceptors, and modulate the overall inotropic response to NA in the adult.

Adrenergic alpha-Agonists↗

PubMed1995

Effect of sympathectomy on inotropic responsiveness to alpha-adrenoceptor stimulation in developing mouse myocardia.

Effects of postnatal sympathectomy on inotropic responsiveness to alpha-adrenoceptor stimulation were examined in mouse myocardia to determine whether the developmental conversion of alpha-adrenoceptor-mediated inotropic responses from positive to negative is triggered by sympathetic innervation. Sympathectomy was performed chemically by consecutively administering 6-hydroxydopamine for 14 days after birth and confirmed by the absence of inotropic responses to tyramine. In newborn myocardia, phenylephrine, in the presence of propranolol, produced concentration-dependent positive inotropic responses. Three weeks after birth, phenylephrine, in the presence of propranolol, produced concentration-dependent negative inotropic responses, both in control and in sympathectomized myocardia; no difference was observed between the two groups of mice in the maximum decrease in contractile force produced by phenylephrine. The sensitivity (pD2 value) to phenylephrine was significantly higher in sympathectomized myocardia. In conclusion, sympathetic innervation of the mouse ventricular myocardium is not required for the developmental conversion of the alpha-adrenoceptor-mediated inotropic response from positive to negative.

Adrenergic alpha-Agonists↗

PubMed1995

Effects of cyclopiazonic acid on rat myocardium: inhibition of calcium uptake into sarcoplasmic reticulum.

The effects of cyclopiazonic acid (CPA), which is reported to inhibit Ca++ uptake into intracellular Ca++ store sites in skeletal and smooth muscles, were examined in ventricular myocardia of the adult rat. In isolated papillary muscle, 0.3 to 30 microM CPA concentration-dependently decreased the contractile force and prolonged the relaxation phase of twitch contractions. In skinned papillary muscle with functional sarcoplasmic reticulum (SR) preserved, application of CPA during the Ca++ loading period reduced the peak tension of subsequent caffeine-induced contractions. When CPA was applied during exposure to caffeine, it had no significant effect on the peak tension, but prolonged the decay of the caffeine-induced contraction. CPA had no substantial influence on the pCa-tension relationship of skinned fibers without functional SR. CPA had no substantial influence on the peak inward and steady-state membrane currents of isolated myocardial cells. These results suggest that CPA inhibits Ca uptake into SR by acting as a specific inhibitor of SR Ca(++)-ATPase in cardiac muscle as in skeletal and smooth muscles and thus provides a valuable pharmacological tool for studying myocardial excitation-contraction mechanisms.

Animals↗

PubMed1994

Expression and function of multidrug resistance P-glycoprotein in a cultured natural killer cell-rich population revealed by MRK16 monoclonal antibody and AHC-52.

Natural killer (NK) cells have been reported recently to be the highest in expressing multidrug resistance (MDR) P-glycoprotein among normal mature lymphoid cells. Using a cultured NK cell-rich population, we have examined the expression and function of P-glycoprotein, in particular its role in NK cell-mediated cytotoxicity, by employing two MDR-reversing agents (nicardipine and AHC-52, a nicardipine analog almost devoid of calcium channel blocking activity) and monoclonal antibody against P-glycoprotein (MRK-16). The expression of P-glycoprotein was detected by flow cytometry and polymerase chain reaction of reverse transcribed mRNA. P-glycoprotein was functional in terms of rhodamine dye excretion and its susceptibility to the MDR-reversing agents. Since the concentration of nicardipine required for 50% inhibition (IC50) of rhodamine dye excretion (2 microM) was close to that of AHC-52 (5 microM), it was suggested that their inhibitory effects were not due to calcium channel blocking activity, and that ACH-52 is a selective inhibitor for P-glycoprotein. The IC50 of nicardipine for NK cell-mediated cytotoxicity (33 microM) was also close to that of AHC-52 (26 microM), indicating that P-glycoprotein is involved in NK cell-mediated cytotoxicity. In support of this, MRK16 inhibited NK cell-mediated cytotoxicity in a concentration-dependent manner. Both binding of target cells to NK cells and post-binding events were affected by AHC-52, suggesting that P-glycoprotein is involved in several steps in NK cell-mediated cytotoxicity.

ATP Binding Cassette Transporter, Subfamily B, Mem↗

PubMed1994

Inotropic effects of ryanodine and nicardipine on fetal, neonatal and adult guinea-pig myocardium.

The effects of extracellular Ca2+ and inotropic agents on contractile force were examined in left atrial and right ventricular myocardia isolated from fetal (35-55 days after conception), neonatal (0-5 days after birth) and adult (30-90 days after birth) guinea-pigs. In both atrial and ventricular muscles, the contractile force increased with increasing extracellular calcium concentration ([Ca2+]o) and the sensitivity to [Ca2+]o was higher in the fetus than in the neonate and adult. Nicardipine almost abolished the contractile force in all groups examined. The sensitivity to nicardipine was similar among the three age groups and also between the two tissues. Ryanodine almost abolished the contractile force in atrial muscles from all age groups. In ventricular muscles it partially reduced the contractile force to the same extent in the neonate and adult while it slightly increased the contractile force in the fetus. In both atrial and ventricular muscles, isoproterenol shortened the relaxation time to the same extent in the neonate and adult, but little or not in the fetus. Taken together, our present results suggest that the sarcoplasmic reticulum function in regulating myocardial contraction is less developed in the fetal guinea-pig when compared with the neonate and adult, and that the atrial contraction is more dependent on Ca2+ release from the sarcoplasmic reticulum than the ventricular contraction is. Thus, it appears that the development of contractile function of the guinea-pig myocardium is mostly completed by birth.

Animals↗

PubMed1994

Differential effects of hypoxia on electrical and mechanical activities of isolated ventricular muscles from fetal and adult guinea-pigs.

1. Effects of hypoxia (95% N2 + 5% CO2) and cromakalim (30 microM) on mechanical and electrical activities of isolated ventricular muscles were examined in fetal and adult guinea-pigs. 2. Hypoxia markedly reduced both the action potential duration (APD) and the contractile force (CF) in the adult, which was only partially restored by 10 microM glibenclamide, while it only slightly reduced CF with little affecting APD in the fetus. 3. Cromakalim markedly reduced both APD and CF in both age groups similarly. 4. Thus, we demonstrated that APD and CF of fetal ventricular muscles are resistant to hypoxia as compared with those of adult. This may be explained by difference in metabolic pathways rather than by lack of ATP-sensitive potassium channels.

Action Potentials↗

PubMed1994

Post-natal decrease in chronotropic sensitivity to acetylcholine in rat heart.

1. The negative chronotropic effects of acetylcholine and carbachol on isolated rat right atria were examined at 0, 4, 8, and 16 weeks after birth. 2. Acetylcholine produced negative chronotropic responses at all ages and completely abolished spontaneous beating at its maximum effective concentration. 3. The sensitivity to acetylcholine, expressed in terms of ED50 values, was higher at 0 and 4 weeks than at 8 and 16 weeks, ED50 values (microM) at 0, 4, 8 and 16 weeks being 9.5 +/- 1.8 (n = 12), 13.2 +/- 3.4 (n = 11), 59.3 +/- 10.9 (n = 14) and 51.5 +/- 17.5 (n = 5), respectively. 4. Neostigmine produced a leftward shift of the concentration-response curve for acetylcholine both at 4 and 8 weeks after birth. The shift was larger at 8 weeks and no difference in sensitivity to acetylcholine was observed between the two ages in the presence of neostigmine. 5. Further, no developmental changes were observed in the sensitivity to carbachol, which is not hydrolyzed by cholinesterase. 6. We concluded that the chronotropic sensitivity to acetylcholine of rat atria decreases post-natally during the period between 4 and 8 weeks after birth due to increase in cholinesterase activity.

Acetylcholine↗

PubMed1994

Effects of low sodium ouabain and amiloride on chick myocardial contraction.

1. Effects of Ca2+, low Na+, ouabain and amiloride on contractile force were examined in myocardial preparations from embryonic and hatched chicks. Measurement of contractile force was performed in an organ bath with whole hearts for the young embryo (5-6 days old) and with isolated strips from the right ventricles for the old embryos (16-18-day-old), hatched chicks (within 24 hr after hatching) and 1-week-old chicks. 2. The maximum increase in contractile force above the basal value at each age was determined by increasing extracellular Ca2+, which was smaller in young embryonic hearts than in older ones. 3. Decreasing the Na+ concentration to half of the normal value increased the contractile force in all ages examined: the increase in contractile force expressed as a percentage of the maximum increase at each age was significantly larger in young embryonic hearts than in older ones. 4. Ouabain produced increases in contractile forces at all ages examined; the effective concentration was lower in young embryonic hearts than in older hearts. 5. Amiloride prolonged the relaxation phase of contraction at all ages examined; the degree of prolongation was larger in young embryonic hearts than in older ones. 6. The present results suggest that, in the chick myocardium, the relative role of Na(+)-Ca2+ exchange in relaxation decreases during embryonic development, which may reflect the concurrent increase in sarcoplasmic reticulum function [Tanaka, Takagi and Shigenobu (1993) J. Dev. Physiol. 19, 235-240.

Amiloride↗

PubMed1994

Myocardial action potential prolongation by calcium channel activation under calcium-free EGTA condition in guinea pigs and rats.

1. Prolongation of action potentials upon the addition of isoproterenol or dbcAMP under Ca-free EGTA condition were observed in isolated myocardial preparations from both the guinea pig and the rat, whose action potential configuration greatly differ. The degree of prolongation was greater in the rat than in the guinea pig. 2. The prolongation of the action potential was rapidly reversed upon the addition of calcium ion and was dose-dependently suppressed by the addition of calcium antagonists. The sensitivity to nicardipine of this action potential was tenfold higher than of the so-called slow response action potentials. The duration of the prolonged action potential was dependent on the external sodium concentration, but was not affected by tetrodotoxin. 3. Thus, it was demonstrated in intact myocardia that sodium ion may persistently pass through the calcium channel to prolong the action potential when it is activated under the condition where the calcium-mediated inactivation of calcium channels is removed. 4. Contribution of calcium-mediated inactivation of calcium channels to the repolarization of normal myocardium may be larger in the rat than in the guinea pig.

Action Potentials↗

PubMed1994

Increased sensitivity of neonatal mouse myocardia to autonomic transmitters.

Chronotropic and inotropic responses to noradrenaline and acetylcholine were examined in isolated right atrial and ventricular preparations from neonatal and adult mice. Noradrenaline and acetylcholine produced positive and negative chronotropic responses, respectively, in the atria from both ages. Noradrenaline produced positive inotropic responses in ventricular preparations from both ages. In all cases, the sensitivity, expressed in terms of pD2 values, was higher in neonatal preparations. In the ventricle, desipramine produced a leftward shift of the concentration-response curve for noradrenaline in the adult, but no such shift was observed in the neonate. The sensitivity to isoprenaline of ventricular preparations was higher in the neonate than in the adult. Our results demonstrated developmental decreases in sensitivities to autonomic transmitters in mouse myocardia. As for the inotropic response to noradrenaline of ventricular muscle, both pre- and postjunctional mechanisms were responsible for the developmental decrease in sensitivity.

Acetylcholine↗

PubMed1994

Effects of tetrodotoxin and low-sodium on action potential plateau of ventricular myocardium from neonatal and adult guinea-pigs.

We examined the effects of tetrodotoxin and reduced extracellular sodium concentrations on the myocardial action potential plateau in neonatal and adult guinea-pigs. Control action potential duration at 50% repolarization (APD50) was significantly shorter in the neonate than in the adult. Tetrodotoxin and low sodium shortened APD50 more prominently in the adult than in the neonate. Since a late sodium current with slow inactivation kinetics is reported in the ventricular myocytes, the results suggest that the late sodium current might contribute to the development prolongation of the action potential duration observed in the guinea-pig myocardium.

Action Potentials↗

PubMed1993

BMPs induce direct bone formation in ectopic sites independent of the endochondral ossification in vivo.

Bone formation in vivo occurs via two major processes, one of which depends on pre-existing cartilage, and the other does not. Bone morphogenetic proteins (BMPs) have been suggested to induce cartilage formation from non-skeletogenic mesenchymal cell population, which results in osteogenesis through the endochondral sequence. In the present study we examined if BMPs could cause direct bone formation independent of pre-existing cartilage using bovine fibrous collagen membrane (FCM) as a carrier for BMPs. Bovine metatarsal bone was extracted in 4 M guanidine HC1 and BMPs were partially purified through the hydroxyapatite chromatography and the Heparin-Sepharose CL6B chromatography. The carrier was loaded with BMPs and then implanted in Wistar rats subcutaneously. The implants were fixed together with surrounding tissue every week after implantation and processed for von Kossa stain, immunohistochemistry, and electron microscopy. The phenotypes of bone and cartilage were identified histologically and immunohistochemically using antibodies against type I and type II collagen. Cartilage and bone were independently induced by 2 weeks. The bone formed directly on the collagen substrate of FCM without pre-existing cartilage. Calcification occurred in the carrier as well as the cartilage and bone matrix. The present study suggests that the BMPs induce osteogenesis in vivo independent of the endochondral sequence.

Animals↗

PubMed1993

Positive chronotropic and inotropic responses to lysophosphatidylcholine are mediated by norepinephrine released from myocardial sympathetic nerve terminals.

1. The effects of beta-adrenoceptor blockade and reserpinization on the positive chronotropic and inotropic responses to lysophosphatidylcholine (LPC) were examined in isolated atrial and ventricular preparations from rat hearts. 2. The positive responses to 10(-4) M LPC were about 40-55% of those to 10(-5) M norepinephrine (NE) in each preparation. 3. The responses to LPC in the presence of propranolol or in the reserpinized preparations were significantly smaller than the corresponding values obtained in control preparations, and were about 10-25% of those to NE. 4. It was concluded that positive chronotropic and inotropic responses to LPC are at least partially mediated by release of NE from myocardial sympathetic nerve terminals.

Animals↗

PubMed1993

Hypotensive effect of bopindolol in pithed rats.

1. Effects of bopindolol, a beta adrenoceptor antagonist with partial agonist activity, on the diastolic blood pressure and heart rate of pithed rats were compared with those of pindolol. 2. Both bopindolol and pindolol reduced the diastolic blood pressure (DBP) in pithed rats. Bopindolol (3.0 mg/kg) and pindolol (1.0 mg/kg) produced similar decreases in DBP of about 8 mmHg; thus pindolol had a 3-fold higher hypotensive potency when compared with bopindolol. 3. Both drugs also produced a dose-dependent decrease in heart rate. 4. Propranolol (5 mg/kg) had no antagonistic effect on the decrease in DBP produced by either bopindolol or pindolol. 5. These results suggest that both bopindolol and pindolol reduce DBP and heart rate in pithed rats through some mechanism independent of beta-adrenoceptors.

Adrenergic beta-Antagonists↗

PubMed1993

Studies on the hypotensive mechanisms of NZ-105, a new dihydropyridine derivative, in rats and rabbits.

1. Intravenous administration of NZ-105 caused a slow-onset and long-lasting hypotension in anesthetized SHR. 2. Centrally administered NZ-105 (0.04 mg/kg) slightly decreased blood pressure. 3. The hypotension of NZ-105 (0.1 mg/kg, i.v.) was not affected by atropine, propranolol, diphenhydramine plus cimetidine, aminophylline or indomethacin. 4. In ganglion-blocked rats, NZ-105 (0.003-0.3 mg/kg, i.v.) inhibited the pressor response to several hypertensive agents to a similar degree. 5. In pithed SHR, NZ-105 (0.03 mg/kg, i.v.) showed the same degree of hypotensive action as in non-treated SHR. 6. NZ-105 did not inhibit reflex responses to tilting in conscious rabbits. 7. Thus NZ-105 exerts its hypotensive action through the mechanisms of peripheral origin.

Animals↗

PubMed1993

Possible action of cyclopiazonic acid on myocardial sarcoplasmic reticulum: inotropic effects on neonatal and adult rat heart.

Cyclopiazonic acid (CPA), a mycotoxin from Aspergillus and Penicillium, has been described as a highly selective inhibitor of Ca(2+)-ATPase in the sarcoplasmic reticulum (SR) in skeletal and smooth muscles but no reports at present deal with the effect of CPA in cardiac muscle. In the present study, we examined the inotropic effect of CPA on adult and neonatal rat myocardia, the contractions of which are known to be highly dependent on Ca(2+)-release from the sarcoplasmic reticulum and transsarcolemmal Ca(2+)-influx, respectively. CPA (30 microM) produced a negative inotropic effect in adult preparations, accompanied by marked prolongation of the contraction duration. In contrast, CPA had minimum effects on neonatal myocardium. Thus we have demonstrated that CPA exerts negative inotropic effects on adult myocardium probably through inhibition of SR function.

Aging↗