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Myocytes were enzymatically isolated from large epicardial arteries of the pig. In the cell attached configuration, we studied currents through L-type Ca-channels. At 22°C, open channel conductance was 9 pS with 110 mM Ba²⁺ and 24 pS with 110 mM Ba²⁺ as charge carrier. According to the life time of the open state, 2 ’modes’ of gating are distinguished; mode 1 contributed time constants shorter than 1 ms, mode 2 those longer than 6 ms to the open time distribution. Mode 2 openings appeared spontaneously, more frequently with Ba²⁺ than with Ca²⁺ as charge carrier. The Ca-agonist Bay K 8644 (0.5 pM) facilitated the appearance of mode 2. Bath application of the phenylalkylamine D600 (1 µM) did not change the gating modes, but it reduced the channel openness by increasing the percentage of blank records. With whole cell recordings, we studied reduction of Ica by 1 µM D 600 at 3.6 mM [Ca²⁺] and 35°C. At a holding potential of -45 mV, D600 induced an ’initial block’ of 35% (10% at -65 mV). Upon repetitive 1 Hz pulsing (170 ms to 0 mV) an additional, ’use-dependent’ block developed with time. More negative holding potentials attenuated reduction of Ica by D600, hyperpolarizations to -100 mV had an ’unblocking’ effect. In regard to reduction of Ica, we compared the partially uncharged D 600 (membrane permeable) with the completely charged compound D890 (membrane impermeable). When applied with the bath, 1 or 10 pM D 600 reduced Ica dose-dependently whereas D 890 was ineffective. When D890 was applied via the patch electrode to the cytosol, it reduced Ica. We discuss that D 600 enters the cell in the uncharged lipid soluble form and reaches form the inside its receptor associated with the Ca-channel.
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Effects of a bradycardic agent on postischemic cardiac recovery in rabbits

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Decreasing heart rate might be beneficial for improvement of myocardial energetics and could reduce the severity of myocardial ischemia. We examined the contribution of heart rate reduction by cilobradine (DK-AH 269), a direct sinus node inhibitor, on left ventricular function and peripheral vasomotion in anesthetized rabbits with experimental myocardial infarction. The rabbits were randomized to receive either placebo (n=10) or cilobradine (n=7). Cilobradine decreased significantly heart rate from 163 ± 33 to 131 ± 13 bpm, p< 0.05, without any inotopic or vascular effects. After 60 min coronary occlusion and 30 min reperfusion, both systolic and diastolic ventricular function were more reduced in the cilobradine group; i.e. maximal left ventricular pressure significantly decreased to 62 ± 11 mmHg, p < 0.05 (placebo: 77 ± 9 mmHg); dP/dtmin significantly decreased to -904 ± 247 mmHg, p < 0.05 (placebo: -1106 ± 242 mmHg). However, infarct size in the cilobradine group was significantly smaller compared with the placebo group. In conclusion, cilobradine reduced heart rate without any negative inotropic effect and reduced infarct size. On that account, this bradycardic agent might open a promising therapeutical avenue to treat postischemic dysfunction.
Badano wpływ amlodypiny, diltiazemu, nifedypiny i verapamilu na antynocyceptywne działanie Padmy 28 w teście wicia się u myszy. Wykazano, że Padma 28 istotnie zmniejsza reaktywność bólową myszy. Antagoniści kanału wapniowego: nifedypina (500 ug/kg i.p) i verapamil (10 ug/kg i.p). nie zmieniały działania, natomiast amlodypina (1 (ig/kg i.p) nasilała, a diltiazem (100 ug/kg i.p) istotnie zmniejszał antynocyceptywne działanie Padmy 28.
The aim of this study was to investigate whether the L-type calcium current (ICa.L) may be altered in aged hearts and whether the classical calcium antagonist verapamil may exhibit altered pharmacological profile in aged hearts. We examined male New Zealand rabbits aged either 6 months or 26 months. To examine ICa.L whole-cell patch-clamp technique was performed on isolated cells. Moreover, activation-recovery intervals (ARI) of isolated hearts (Langendorff method) were assessed using an epicardial 256 channel mapping system. We found that the ICa.L density, normalised to the cell volume was significantly reduced (p<0.001). Maximum conductance was also significantly decreased (p=0.01) and steady state inactivation was shifted to more positive potentials in aged hearts (p<0.001). A slightly reduced effect of ß-adrenergic modulation of the ICa.L in aged hearts, and a significantly reduced effect of carbachol on isoprenaline-stimulated ICa.L in aged hearts was observed. L-type 1c subunit, SERCA2-ATPase and the Na+/Ca2+-exchanger expression were neither significantly different in atrial and ventricular tissues nor between young and old animals. Using the mapping system, isolated hearts were exposed to verapamil (0.005, 0.01, 0.02, 0.05 µM/L). While verapamil did not affect ARI in young hearts, in aged hearts ARI was concentration-dependently reduced and the negative inotropic effect of verapamil was significantly attenuated in aged hearts (p<0.05). From these results we conclude that there are distinct alterations in the electrophysiology of ICa.L (reduced maximum conductance, a shift of the steady state inactivation) in the aged heart which may influence the response to verapamil.
Calcium ionophore A23187 allowing for a calcium ion influx from an apoplast to a cytoplasm, mimicked symptoms of the frost-induced injuries in winter oilseed rape leaves, as estimated by the conductivity method. Both calcium ionophore and freezing treatment induced degradation of phosphatidylcholine. On the other hand lanthanum and gadolinum ions as well as verapamil, the inhibitors of calcium ion channels, decreased the degree of the frost-induced injuries. Lanthanum ions prevented the frost-induced degradation of PC. It is proposed that freezing alters the functioning of calcium ion channels which results in calcium ion influx into a cytosol. This in turn may lead to a degradation of cell membranes.
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