Effects of Bay k 8644 and nifedipine on isolated dog cerebral, coronary and mesenteric arteries

Vasoconstrictor effects of Bay k 8644, a dihydropyridine Ca++ agonist, and vasorelaxant effects of nifedipine were investigated in helical strips of dog cerebral (basilar, posterior cerebral and middle cerebral) and peripheral (coronary and mesenteric) arteries. The addition of Bay k 8644 produced a dose-dependent contraction in the absence of any contractile agent in the basilar artery with a pD2 value of 8.53. Similar sensitivity to Bay k 8644 was observed in the posterior cerebral, middle cerebral or coronary artery. Bay k 8644 was much less effective in producing a contraction in the mesenteric artery. An elevation of the concentration of extracellular K+ eliminated the difference between the responses to Bay k 8644 in the basilar and mesenteric artery. Contractile responses of the basilar artery to Bay k 8644 were antagonized competitively by nifedipine (pA2 = 8.17), but non-competitively by diltiazem. The pA2 values for nifedipine antagonism of Bay k 8644 responses with the elevated K+ were the same between the basilar and mesenteric arteries. Increased sensitivity to exogenously added K+ also was observed in cerebral and coronary arteries when compared with the mesenteric artery. The addition of nifedipine to an unstimulated strip produced a dose-dependent relaxation in cerebral and coronary arteries, but not in the mesenteric artery. When the cerebral and peripheral arteries were contracted with K+ to the same magnitude, nifedipine produced similar relaxations among these arteries. Nifedipine was less efficacious in antagonizing the contractile response to Bay k 8644 compared with the contractile response to K+ in cerebral arteries. These results suggest that 1) the voltage-dependent Ca++ channels in the cerebral and coronary arteries are in different states of activation from those in the mesenteric artery, 2) Bay k 8644 contracts the cerebral and coronary arteries by acting primarily on the same site, presumably dihydropyridine receptors of the voltage-dependent Ca++ channels at which nifedipine acts, 3) the dihydropyridine receptors were the same between the basilar and mesenteric arteries and 4) there may be a difference in the state of the Ca++ channel in the arteries between the stimulation with Bay k 8644 and K+-depolarization.     

Spontaneously hypertensive rat resistance artery structure related to myogenic and mechanical properties

This investigation related arterial structure to myogenic (pressure-dependent) contractile responses in resistance arteries from spontaneously hypertensive rats (SHRs) and Wistar-Kyoto (WKY) normotensive control rats under pressurized conditions in vitro. Femoral and mesenteric resistance arteries from either strain were cannulated and pressurized in an arteriograph for the determination of pressure-diameter relationships under passive and active conditions in the range 5-200 mmHg transmural pressure. Arterial geometrical measurements were made under relaxed conditions at 100 mmHg. Media thickness/lumen diameter (M/L) ratios were significantly increased in SHR femoral (5.00+/-0.44% compared with 3.63+/-0.34%; P<0.05) and mesenteric (4.40+/-0.29% compared with 2.62+/-0.23%; P<0.001) arteries compared with those from WKY rats. Maximum myogenic contractions, assessed as minimum normalized diameters, were not significantly different in SHR and WKY rat femoral (0.41+/-0.03 and 0.40+/-0.02 respectively) or mesenteric (0.56+/-0.02 and 0.63+/-0.03 respectively) arteries. Arterial mechanical analyses demonstrated that incremental elastic modulus is reduced in SHR mesenteric arteries, but is not significantly different in SHR femoral arteries, compared with those from WKY rats. Additionally, wall stress at estimated in vivo pressures under passive and active conditions are similar in SHR and WKY rat arteries. These data demonstrate that increased M/L ratios in resistance arteries from SHRs are not associated with increased maximum pressure-dependent contractile responses. Increased M/L ratios in resistance arteries from SHRs are not accounted for by increased vessel wall stiffness, but the hypertension-associated arterial geometrical abnormalities act to normalize wall stress in the face of increased arterial pressure.     

Acute and chronic captopril, but not prazosin or nifedipine, normalize alterations in adrenergic intracellular Ca2+ handling observed in the mesenteric arterial tree of spontaneously hypertensive rats

The effect of hypertension and acute (36-h) or chronic (from age 6 to 16 weeks) antihypertensive treatment with prazosin (2 mg kg(-1) per day), nifedipine (50 mg kg(-1) per day), or captopril (50 mg kg(-1) per day) on Ca2+ mobilization due to alpha1-adrenoceptor activation was analyzed in functional studies using arterial rings [four conductance/distributing vessels: aorta, main mesenteric, iliac, and tail arteries and two resistance vessels; first and second small mesenteric artery branches obtained from spontaneously hypertensive rats (SHR, 6 and 16 weeks old) and age-matched Wistar Kyoto rats (WKY)]. Maximal response to noradrenaline in the presence of extracellular Ca2+ is not affected by hypertension or by the antihypertensive treatment. The extracellular Ca2+-independent contractile responses increased with age in iliac, tail, and small mesenteric arteries (SMA) and were further increased in SHR in SMA from both young and adult animals and in the main mesenteric artery of adult SHR. In main mesenteric artery, this increased contraction in SHR was associated with a higher increase in cytosolic [Ca2+] mobilized by noradrenaline without changes in the total stored Ca2+. Acute or chronic treatment with captopril abolished the differences observed between WKY and SHR in the noradrenaline-induced contraction in mesenteric arteries loaded in Ca2+-free medium. In contrast, animals acutely treated with prazosin or chronically treated with either prazosin or nifedipine exhibit the same differences in Ca2+ handling than untreated rats. In conclusion, these differences are not a consequence of increased blood pressure but precede it and can only be normalized by inhibition of the rennin-angiotensin system.     

Agonist actions of Bay K 8644, a dihydropyridine derivative, on the voltage-dependent calcium influx in smooth muscle cells of the rabbit mesenteric artery

Actions of methyl-1,4-dihydro-2,6-dimethyl-3-nitro-4-(2- trifluoromethylphenyl)-pyridine-5-carboxylate (Bay K 8644) on the mechanical response evoked in intact and skinned mesenteric artery of the rabbit were investigated. The data were compared to that of nisoldipine, another dihydropyridine derivative Bay K 8644 increased the amplitudes of both the phasic and tonic components of the K+-induced contraction which is due to an increase in the voltage-dependent influx of Ca ion. Bay K 8644 antagonized competitively the actions of nisoldipine (a Ca antagonist) on the tonic but not on the phasic component of the K+-induced contraction. The contractions caused by high concentrations of norepinephrine were enhanced to a greater extent by Bay K 8644 than that evoked by lower concentrations of norepinephrine. Bay K 8644 had no effect on Ca++ extrusion from cells, which was estimated from the change in amplitudes of the norepinephrine-induced contractions in Na+- and Ca++-free solutions. This agent had no effect on the contractile proteins and Ca storage sites, as estimated from the Ca++- or caffeine-induced contraction observed in skinned muscles. The results suggested that Bay K 8644 acts primarily on the voltage-dependent Ca++ channel, presumably the same site at which other dihydropyridine derivatives (Ca antagonists) act, and that the influx of Ca++ is accelerated.     

Influence of experimental reduction of media/lumen ratio on arterial myogenic properties of spontaneously hypertensive and Wistar-Kyoto rats

In the present study, myogenic properties of femoral arteries from control hindlimbs and those distal to external iliac artery partial ligation of spontaneously hypertensive rats (SHRs) and normotensive Wistar-Kyoto (WKY) rats were assessed. Arterial pressure was reduced distal to the ligature in both strains. Media thickness/lumen diameter (M/L) ratios of control (unligatured) SHRs were greater than in unligatured WKY rats and were reduced in arteries distal to the ligature (ligatured) within each strain. In none of the comparisons was a greater M/L ratio associated with greater maximal myogenic contractions, but increased M/L ratios were associated with a shift of myogenic activity to a higher pressure range in all comparisons. SHR ligatured arteries produced greater pressure-dependent contractile responses than WKY rat unligatured arteries, although arterial structures were not significantly different. Wall stress was similar in all arteries within the 60-120 mmHg pressure range with myogenic tone in spite of large differences in arterial structure. The utilization of arteries with experimentally altered structure provides further evidence that increased M/L ratios are not associated with greater peak pressure-dependent contractile responses and that arterial wall stress is maintained within a narrow range through an interaction between arterial wall geometry and smooth muscle contractile function.     

Responses of arterial smooth muscle from normotensive and pre-eclamptic subjects to the calcium channel agonist,Bay K 8644

The effect of Bay K 8644, a dihydropyridine Ca2+ agonist, on in vitro contractile responses of inferior epigastric arteries from normotensive (N) and pre-eclamptic (P) subjects has been investigated, with a view to further defining the mechanism of the increased vascular sensitivity associated with pregnancy-induced hypertension. Bay K 8644 (10(-10)-10(-7) M) caused dose-dependent contractions of N as well as P arteries under resting conditions in the order: P greater than N and caused development of rhythmic contractions in both N and P arteries. Bay K 8644 effects were prevented by 3 X 10(-8) M Nifedipine (a Ca+2 antagonist). Bay K 8644 also significantly (P less than 0.05) enhanced the sensitivity as well as maximal contractile responses to CaCl2 in 40 mM K+-depolarized Ca-depleted N and P arteries in the order: P greater than N. The results suggest that the increased peripheral vascular sensitivity associated with pregnancy-induced hypertension may be due, at least in part, to enhanced activity of the potential-sensitive Ca2+ channels in arterial smooth muscle plasmalemma.     

Is antagonism by Bay k 8644 of the negative dromotropic effect of nifedipine pharmacological?

The dromotropic effect of Bay k 8644 and its interaction with the negative dromotropic effects of nifedipine, verapamil, MnCl2 and tetrodotoxin were investigated by use of isolated, blood-perfused atrioventricular (AV) node preparations of dogs. These agents were injected into the AV node artery. Single injections of Bay k 8644 (0.1-10 micrograms) shortened AV conduction time, but the decrease remained only about 9 msec at 10 micrograms. This effect was not modified by nadolol. Dose-response curves for the negative dromotropic effect of nifedipine were shifted to the right by about 0.5 log units with infusions of Bay k 8644 into the AV node artery at rates of 3 and 10 micrograms/min. A similar shift of dose-response curves to verapamil occurred at 10 micrograms/min of Bay k 8644. However, Bay k 8644 (3 and 10 micrograms/min) failed to modify dose-response curves to MnCl2 and tetrodotoxin. Bay k 8644 (10 micrograms) produced a greater decrease in AV conduction time during the negative dromotropic effect of verapamil (10 or 30 micrograms) and a far greater decrease during the negative dromotropic effect of nifedipine (3 or 10 micrograms) than under control conditions. In contrast, Bay k 8644 (10 micrograms) produced a decrease in AV conduction time nearly to the same extent during the negative dromotropic effect of MnCl2 (10 or 30 mumol) or tetrodotoxin (10 or 30 micrograms) as under control conditions. From these results Bay k 8644 can be described as a rather specific pharmacological antagonist for dihydropyridine slow channel blockers.     

Role of endothelium on the effects of neuropeptide Y in mesenteric resistance arteries of spontaneously hypertensive and Wistar-Kyoto normotensive rats

The role of the endothelium in the effects of neuropeptide Y (NPY) and norepinephrine was investigated in mesenteric resistance arteries of the spontaneously hypertensive rat (SHR) and of the normotensive Wistar-Kyoto rat (WKY). Endothelium-dependent relaxation to acetylcholine (1 microM) was reduced in arteries of SHR compared with WKY. In the presence of the endothelium, the vessels of the two strains responded similarly to norepinephrine and NPY (100 nM) produced only a slight contraction. After removal of the endothelium, the response to norepinephrine was greater in WKY than in SHR. Furthermore, endothelium denudation enhanced markedly contraction elicited by NPY in WKY (up to 40% of the maximal effect of norepinephrine), but not in SHR. NPY potentiated the contractile response to low concentrations of norepinephrine (less than 300 nM) in both strains regardless whether the endothelium was intact or not. These results indicate that the contractile responses to NPY and to norepinephrine are inhibited by the endothelium in vessels of WKY, but not in those of the SHR. Furthermore, the potentiating effect of NPY occurs via an endothelium-independent mechanism in mesenteric arteries of both SHR and WKY. It is proposed that the differential responses between the two strains are related to abnormal function of the endothelium and to decreased responsiveness of smooth muscle cells in mesenteric resistance arteries of SHR compared to WKY.     

Plasma membrane calcium flux, protein kinase C activation and smooth muscle contraction

Isolated perfused rabbit ear arteries contract when treated with 12-O-tetradecanoylphorbol-13-acetate (TPA), an activator of the calcium-activated, phospholipid-dependent protein kinase or C-kinase. Under conditions where the calcium concentration in the perfusate is 1.5 mM and the potassium concentration is 4.8 mM, there is a latent period of 70 +/- 19 min (mean +/- S.E.M., n = 10) between TPA addition and the onset of the contractile response. Once initiated, the contractile response is progressive and sustained. When perfusion conditions are altered in such a way as to modify calcium flux across the plasma membrane (i.e., raising the extracellular calcium concentration to 2.5 mM Ca++, raising the extracellular potassium concentration to 10 mM, and/or preincubating the tissues in media containing 100 nM Bay K 8644, a potent calcium channel agonist), the latency period between TPA addition and initiation of the contractile response is significantly reduced (2.5 mM Ca++, 37 +/- 7 min; 10 mM K+ and 2.5 mM Ca++, 11 +/- 3 min; 100 nM Bay K 8644 and 1.5 mM Ca++, 20 +/- 7 min; 100 nM Bay K 8644 and 2.5 mM Ca2+, 8.5 +/- 1.7 min; 10 mM K+ and 100 nM Bay K 8644, 11 +/- 5 min). Likewise, the combination of 2.5 mM calcium, 100 nM Bay K 8644, and 3.3 microM ouabain results in a contractile response 4.5 +/- 2.0 min after TPA addition (means +/- S.E.M., n = 4). Control tissues (absence of TPA addition) run simultaneously show no contractile responses to the various Ca++ flux regulators even after 90 min of incubation.(ABSTRACT TRUNCATED AT 250 WORDS)     

Reduced function of the stimulatory GTP-binding protein in beta adrenoceptor-adenylate cyclase system of femoral arteries isolated from spontaneously hypertensive rats

Arterial relaxant responses to beta adrenoceptor agonists are decreased in spontaneously hypertensive rats (SHR) when compared with normotensive Wistar-Kyoto rats (WKY). To determine which component of the beta adrenoceptor-adenylate cyclase (AC) system is involved in the decreased beta adrenoceptor responses, effects of two activators of AC-forskolin and cholera toxin and of dibutyryl cyclic AMP (DBcAMP) were compared between the strips of femoral arteries isolated from 13-week-old SHR and age-matched WKY. Arterial relaxant responses to either forskolin, an activator of AC or DBcAMP were not significantly different between the SHR and WKY, whereas the relaxant responses to norepinephrine (NE) via beta adrenoceptors were significantly weaker in the SHR than in the WKY. In the absence of timolol, a beta adrenoceptor antagonist, contractile responses to NE were significantly greater in the SHR than in the WKY. Timolol augmented the contractile responses to NE to a greater extent in the WKY than in the SHR. After the blockade by timolol of beta adrenoceptors, contractile responses to alpha adrenoceptor stimulation with NE were not significantly different between the two strains. The pretreatment of the strips with cholera toxin, an activator of stimulatory GTP-binding protein (Gs), antagonized the alpha adrenoceptor-mediated contractions much greater in the WKY than in the SHR. The alpha adrenoceptor-mediated contractions after the pretreatment with cholera toxin were comparable to the contractile responses to NE determined in the absence of timolol in either the SHR or the WKY. Forskolin and DBcAMP also antagonized the alpha adrenoceptor-mediated contractions. However, these antagonisms were not significantly different between the two strains. The cellular cAMP content in arterial strips after the stimulation with NE was significantly less in the SHR than in the WKY, whereas the cAMP contents were similar in arterial strips from both strains which were stimulated with forskolin. These results suggest that the reduced function of Gs is involved in the abnormality of beta adrenoceptor-AC system in the SHR femoral artery.     

Altered inotropic responses in diabetic cardiomyopathy and hypertensive-diabetic cardiomyopathy.

To understand the mechanisms of diabetic cardiomyopathy and the consequences of combined hypertension and diabetes, cardiac tissue responses to various inotropic agents were measured in experimental diabetes. Streptozotocin was injected into Wistar rats, spontaneously hypertensive rats (SHRs) and Wistar-Kyoto rats (WKYs). Six weeks after the injection diabetic rats showed a subsensitivity to beta adrenergic stimulation in ventricular tissue and a supersensitivity and hyper-responsiveness to Ca++ and alpha adrenergic stimulation (except in WKYs) in ventricular tissues and left atria. A supersensitivity to BAY K 8644 in SHR left atria and a hyper-responsiveness to verapamil in ventricular strips were also noted. These alterations may be due to a change in receptor number or to postreceptor alterations. Diabetic SHRs exhibited greater changes in several of the drug responses (responses to isoproterenol, phenylephrine and BAK 8644) were more hyperlipidemic and had a high mortality as compared with Wistar rats and WKY diabetics. These findings confirm that the combination of hypertension and diabetes results in greater cardiac pathology than is seen with either disease alone.     

Endothelial and smooth muscle properties of coronary and mesenteric resistance arteries in spontaneously hypertensive rats compared to WKY rats

Summary— To investigate if the functional alterations observed in resistance arteries of spontaneously hypertensive rats (SHRs) were also present at the coronary level, in vitro experiments were performed in mesenteric resistance arteries (MRA) and in right (RIC) and left interventricular coronary (LIC) arteries taken from 15–25-week-old SHR and age-matched Wistar Kyoto rats WKYs. Using a passive extension protocol, internal diameters corresponding to 100 mmHg intraluminal pressure (D₁₀₀) were determined and vessels were set up to a normalized internal diameter (0.9 D₁₀₀). SHR mesenteric resistance arteries had a significantly smaller diameter compared to WKY arteries, whereas both types of SHR coronary arteries had a greater diameter compared to those of WKY rats. In arteries in the absence of contracting agonist, nitro-L-arginine (NOLA, 100 μM) induced a progressive rise in basal tone, which could be reversed by subsequent addition of L-arginine (100 μM) but not D-arginine (100 μM). When expressed as percent of maximal contractions induced by agonists (noradrenaline, NA [10 μM] in MRA; serotonin, 5-HT [10 μM], in RIC and LIC), these contractions were significantly stronger in WKY compared to SHR coronary and mesenteric resistance arteries. In NA-precontracted MRA and 5HT-precontracted coronary arteries in the presence of indomethacin (10 μM), the magnitude of acetylcholine-induced maximal relaxations (expressed as percent of maximal contractions induced by agonists) was greater in WKY compared to SHR arteries. After a 30-min incubation period, NOLA (100 μM) completely inhibited relaxations induced by acetylcholine (0.01–10 μM) in all types of precontracted arteries. Subsequent additions of sodium nitroprusside, (SNP, 10 μM) induced complete relaxations in all preparations. These results show that a basal release of NO or NO-like compound by endothelial cells is present in isolated mesenteric resistance and coronary arteries of WKY rats and SHRs. The contribution of endothelium-derived relaxing factor-nitric oxide (EDRF-NO) to arterial tone was lower in MRA compared to coronary arteries in both strains and in SHR compared to WKY arteries. In the SHR preparations, the impaired relaxation induced by acetylcholine appeared to be due to a functional alteration of the endothelium in the presence of normal reactivity of the smooth muscle cells.     

Differential inhibitory effect of neomycin on contractile responses of various canine arteries.

The influence of neomycin on vascular smooth muscle contractile responses was examined in different isolated arterial preparations of the dog. Prior exposure (5 minutes) to 7 mM neomycin decreased contractile responses elicited with norepinephrine (NE, 0.6 muM) or KC1 (K-+, 80 nM) in helical strips of canine aortae (Ao) and femoral (F), carotid (Cd), renal (R), superior mesenteric (Sm), terminal mesenteric (Tm) and coronary (Cr) arteries. Addition of neomycin subsequent to NE-induced contractile responses depressed tension responses of the F, Cd, R, Sm and Tm arteries but had little or no effect on the Ao. However, after contractions had been elicited with K-+, neomycin had no effect on tension responses of the Ao, F, Cd, R or Sm arteries but depressed contractions of the Cr and Tm arterial strips. Preincubation with neomycin (0.7-3.5 mM) produced a concentration-related inhibition of contractile responses elicited in Tm arterial strips by addition of calcium ions (Ca-++; 1.6 mM) to a Ca-++-free depolarizing solution; conversely, subsequent addition of neomycin had no effect on maintained Ca-++ contractures. In constant flow-perfused terminal mesenteric arterial branches, neomycin (0.5-4.0 mM) produced a concentration-related antagonism of pressor responses elicited with NE (1-8 mug) or K-+ (40 mM). The inhibitory action of neomycin on K-+-induced pressor responses was inversely related to the Ca-++ concentration of the perfusion fluid. However, inhibition of NE pressor responses by neomycin did not appear to be related to the Ca-++ concentration. The differential inhibitory action of neomycin on contractions induced by NE and K-+ in various canine arteries suggests that different vascular beds vary in the manner in which Ca-++ is bound and subsequently utilized by stimulatory agents to elicit tension changes.     

An endothelium-dependent contraction induced by A-23187, a Ca++ ionophore in canine basilar artery

In most isolated canine basilar arteries tested, Ca++ ionophore A-23187 induced a small relaxation followed by a transient contraction. Both contraction and relaxation were abolished by removal of endothelium. The endothelium-dependent contraction induced by A-23187 was attenuated by a phospholipase A2 inhibitor (quinacrine), cyclooxygenase inhibitors (aspirin and indomethacin), a thromboxane A2 (TXA2) synthetase inhibitor (OKY-046) and a TXA2 antagonist (ONO-3708). The A-23187-induced contraction was abolished by lowering the Ca++ concentration of medium to 10%, whereas the contraction induced by 9,11-epithio-11,12-methano-TXA2 (STA2) was attenuated slightly by lowering [Ca++]. The A-23187-induced contraction was reduced markedly by nifedipine (10(-9) to 10(-7) M), but the STA2-induced contraction was only attenuated slightly by nifedipine. Bay K 8644 did not affect the A-23187- and STA2-induced contractions. The present experiments demonstrate that A-23187 induced an endothelium-dependent contraction in canine basilar artery, and suggest that Ca++ might play a key role in production of an endothelium-derived contracting factor (probably TXA2).     

Defective modulation of noradrenergic neurotransmission by exogenous prostaglandins in aging spontaneously hypertensive rats

Inhibition of cyclooxygenase enhances mesenteric vascular responses to periarterial (sympathetic) nerve stimulation (PNS) in 16-week-old spontaneously hypertensive rats (SHR), but not in 25-week-old SHR. In contrast, cyclooxygenase inhibition enhances mesenteric vascular responses to PNS similarly in 16- and 25-week-old Wistar-Kyoto normotensive rats (WKY). Thus, the modulation of noradrenergic neurotransmission by endogenous PGs becomes defective as SHR age, whereas in WKY this does not occur. The purpose of this study was to determine to what extent alterations in the concentrations of PGs and/or biological response to PGs contribute to this age/hypertension-related abnormality in SHR. All studies were conducted in the in situ autoperfused rat mesentery, and plasma levels of PGE2 and 6-keto-PGF1 alpha were determined by negative-ion, chemical-ionization, gas chromatography-mass spectrometry after derivatization and clean-up of samples by two thin-layer chromatographic steps. Base-line mesenteric venous plasma levels of PGs were similar in 16-week-old SHR vs. 16-week-old WKY; however, base-line levels of PGE2 were approximately 6-fold greater than base-line levels of 6-keto-PGF1 alpha in both strains. PNS at 7 Hz approximately doubled mesenteric venous plasma levels of PGE2 in both 16-week-old SHR and WKY, but PNS did not increase levels of 6-keto-PGF1 alpha in either strain. Inasmuch as mesenteric venous plasma levels of PGE2 were responsive to PNS, the effect of aging on PGE2 levels was studied. In both strains, the base-line mesenteric venous plasma levels of PGE2 and the PNS-induced increase in PGE2 levels were similar in 16-week vs. 25-week-old animals. In 16-week-old SHR, infusions of PGE2, arachidonic acid and PGI2 directly into the mesenteric artery inhibited vascular responses to PNS. However, in 25-week-old SHR, even high doses of PGE2 or arachidonic acid failed to inhibit vascular responses to PNS, and the inhibitory potency of PGI2 was shifted 10-fold to the right compared to 16-week-old SHR. In contrast, PGE2 and arachidonic acid had similar effects on neurotransmission in 25-week-old WKY compared to 16-week-old WKY, and aging had a lesser effect on the inhibitory potency of PGI2 (i.e., 3-fold rightward shift of the dose-response curve). Adenosine also inhibited vascular responses to PNS; however, the inhibitory potency of adenosine was only slightly and similarly affected by aging in SHR and WKY.(ABSTRACT TRUNCATED AT 400 WORDS)     

Vascular and cardiac effects of a new dihydropyridine derivative, YC-170: a comparison with Bay K 8644

The pharmacological effects of YC-170, a new dihydropyridine derivative, were studied in the rabbit aortic strips and guinea pig cardiac preparations and compared with those of Bay K 8644. In the rabbit aortic strips, YC-170 produced contraction in normal physiological saline solution ([K+]0 = 5.9 mM) in a concentration-dependent manner. Increasing the [K+]0 of the medium to 15 mM enhanced the contractile response. The maximum contraction produced by YC-170 at [K+]0 of 15 mM was comparable to that by Bay K 8644. However, YC-170 induced relaxation when the strip was contracted by 60 mM K+. In guinea pig left atrium, YC-170 produced a positive inotropic effect in a concentration-dependent manner, but its extent was far less than that of Bay K 8644. Like Bay K 8644, however, YC-170 increased the time to peak tension and relaxation time of the isometric tension, and prolonged the action potential duration. YC-170 failed to produce a positive inotropic action in the papillary muscle in which Bay K 8644 was a potent positive inotropic agent. In spontaneously beating right atria, YC-170 caused a negative chronotropic effect, whereas Bay K 8644 a positive one. The positive inotropic and vasoconstrictor effects of YC-170 were antagonized competitively by a Ca++ antagonist nicardipine. When the left atria were depolarized with high-K+ medium, the positive inotropic effect of YC-170 was attenuated progressively with increasing [K+]0 and at 13.2 mM K+ a negative inotropic effect was induced by YC-170.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effects of pimobendan, a cardiotonic and vasodilating agent with phosphodiesterase inhibiting properties, on isolated arteries and veins of rats.

Effects of pimobendan (PBD) were investigated on isolated rat blood vessels. PBD dose-dependently relaxed aortic, femoral arterial and mesenteric venous preparations precontracted with KCl and reduced the amplitude of spontaneous contractions of portal venous preparations; the sensitivity to PBD was femoral greater than portal greater than mesenteric greater than aorta. Relaxation response of the femoral artery to PBD was not changed by propranolol and aminophylline. Glyceryl trinitrate (GTN), isoproterenol (ISO), forskolin and adenosine also elicited dose-dependent relaxations of femoral arteries; the rank order of potency (mean negative log EC50 value) was GTN greater than ISO greater than PBD = forskolin greater than adenosine. The relaxation responses to PBD and isobutyl methylaxanthine (IBMX) were not attenuated with removal of endothelial cells. In the femoral artery, methylene blue diminished GTN-induced relaxation but not PBD-induced relaxation. PBD and IBMX increased the relaxation responses of the artery to cyclic AMP-forming drugs (ISO, forskolin and adenosine) but not a cyclic GMP-forming drug (GTN). PBD and IBMX increased the relaxation response of mesenteric veins to ISO. The drugs noncompetitively inhibited arterial contractions accompanied by voltage-dependent and alpha-adrenoceptor-operated Ca2+ influxes. In the absence of extracellular Ca2+, PBD and IBMX reduced contractile responses of arteries to norepinephrine but not caffeine. The present results suggested that PBD relaxed the blood vessels, at least in part, through an intracellular accumulation of cyclic AMP.     

CENTRAL CARDIOVASCULAR EFFECTS OF DIHYDROPYRIDINES IN SPONTANEOUSLY HYPERTENSIVE RATS

Summary— Intracerebroventricular (i.c.v.) injections of dihydropyridine derivatives calcium channel agonist (BAY K8644) and antagonists (nifedipine, nicardipine, PN 200–110) induced opposite long-lasting changes in blood pressure (BP) in pentobarbital anesthetized spontaneously hypertensive rats (SMR). I.c.v. nifedipine (NIF), nicardipine (NIC), and PN 200–110 decreased mean blood pressure dose-dependently and stereoselec-tively, (+) NIC and (+) PN being 8 and 3 times more potent than their (-) isomers, respectively. The decrease in BP was due to a withdrawal of the sympathetic tone, since NIF- and NIC-induced falls in BP were suppressed after either hexamethonium (HXM), 6 OHDA or bilateral adrenalectomy. I.c.v. BAY K8644 increased BP dose-dependently. The i.c.v. BAY K8644-induced hypertensive effect was inhibited: a), by NIF and (+) PN but not by (-) PN, therefore probably occurring at central DHP sites; b), by HXM and reserpine, thus probably mediated by an increase in sympathetic tone; c), by i.c.v. methylatropine (MA) while i.v. MA and i.c.v. HXM had no inhibitory effect, thus probably involving central muscarinic sites. In SHR, NIC did not after the K+-evoked ACh release but suppressed the BAY K8644-induced increase in ACh release. In anesthetized normotensive control rats (WKY), neither i.c.v. NIF, NIC or BAY K8644 changed BP, nor did the latter after ACh release. Moreover, in conscious WKY, i.c.v. nicardipine increased BP and HR while, in conscious SHR it decreased BP without any change in HR. These data suggest that central DHP sites may be involved in the cholinergic transmission and may participate in genetic hypertension via sympathetic tone.     

Role of protein kinase C in electrical-stimulation-induced neuronal nitric oxide release in mesenteric arteries from hypertensive rats

This study examines the influence of hypertension on neuronal nitric oxide (NO) release and its modulation by protein kinase C (PKC). For this purpose, mesenteric segments without endothelium were obtained from Wistar-Kyoto (WKY) rats and spontaneously hypertensive rats (SHRs), and neurogenic NO release induced by electrical field stimulation (EFS) was examined in these segments. EFS induced frequency-dependent contractions. The NO synthase inhibitor N(G)-nitro-L-arginine methyl ester (L-NAME) and the sensorial neurotoxin capsaicin increased EFS-induced contractions in SHR segments, but did not affect these contractions in segments from WKY rats. In segments from SHRs, the increase in EFS-induced response by capsaicin was further increased by the combination of capsaicin and L-NAME. EFS-induced contractions in SHR arteries were unaltered by the protein synthesis inhibitor cycloheximide or by 2-amine-5,6-dihydro-6-methyl-4H-1,3-tiazine (AMT), an inhibitor of inducible NO synthase, and increased by the guanylate cyclase inhibitor Methylene Blue. In these arteries, capsaicin plus the PKC inhibitor calphostin C increased the contractions elicited by EFS; the addition of L-NAME did not affect this increase. Phorbol 12,13-dibutyrate (PDBu) did not modify the response to EFS in these arteries pretreated with capsaicin, although a combination of PDBu and L-NAME was effective. These results indicate that, in mesenteric arteries, EFS induces the release of NO from perivascular nitrergic nerves and of neuropeptides from sensory nerves, but only in hypertensive rats. The NO released is synthesized by constitutive neuronal NO synthase in a manner that is positively modulated by PKC, an enzyme that seems to be activated in hypertension.     

Analysis of the interaction between lacidipine and BAY K 8644 in two isolated rabbit arteries.

The calcium antagonist activity of the long-acting 1,4-dihydropyridine (DHP) lacidipine has been analyzed in rabbit ear artery (REA) and rabbit basilar artery (RBA). Its potency has been estimated from its interaction with BAY K 8644 using a three state gating model of the voltage-operated calcium channel. As a contractile agent, BAY K 8644 exhibited a bell-shaped concentration-response curve in both arteries. For fitting purposes, a second binding interaction between BAY K 8644 and the channel has been used to describe the descending part of the curve. The K(app)s for lacidipine and three other DHPs (nifedipine, nitrendipine and amlodipine) have been compared to pA2 values obtained from displacement of calcium concentration-response curves. In both REA and RBA the K(app)s for the four DHPs were not significantly different compared to their pA2s. The pK(app) values for lacidipine were estimated as 9.80 for REA and 10.20 for RBA.