Spironolactone inhibition of contraction and calcium channels in rat portal vein.
1. The effects of spironolactone have been studied on the mechanical activity of rat portal vein strips and the calcium channel currents of isolated cells using the patch clamp technique (whole-cell configuration). 2. Spironolactone (50 nM to 0.1 mM) depressed both K+-induced and twitch contractions within 5-6 min. This inhibitory effect was overcome by elevating the calcium concentration in the perfusing solution. 3. Spironolactone (60 microM) depressed the transient contractions induced in a Ca2+-free, EGTA-containing solution by either acetylcholine (0.1 mM) or noradrenaline (10 microM). The effect of spironolactone was dependent on a reduction in the filling of the internal calcium store. 4. Rapidly inactivating calcium channel current was maintained in the presence of spironolactone (60 microM), while slowly inactivating calcium channel current was blocked in a concentration-dependent manner. Half-inhibition of slow calcium channel current was obtained at concentrations between 5-7 microM. 5. Administration of spironolactone (10 microM) at rest reduced calcium channel current by about 70% (tonic inhibition). Repetitive depolarizations (300 ms long pulses to zero mV, applied between 0.05 and 0.5 Hz) had no further inhibitory effect on the inward current (absence of use-dependence). 6. When cells were held at depolarized membrane potentials at which slow calcium current was inactivated by about 80%, the inhibitory effect of spironolactone (10 microM) was similar to that obtained with cells normally polarized. Spironolactone (10 microM) had no effect on the voltage-dependence of inactivation of the calcium channel current. 7. Our results suggest that spironolactone acts primarily on the plasma membrane by depressing inward current through slow calcium channels. This effect may be explained by a preferential binding of the drug to the resting state of the slow calcium channel. In addition, spironolactone may depress contractions dependent on the release of calcium from the sarcoplasmic reticulum.
Institut de Biochimie et Neurochimie du C.N.R.S., Université de Bordeaux II, France.
C
C
Dacquet
G
G
Loirand
C
C
Mironneau
J
J
Mironneau
P
P
Pacaud
D000818:Animals; D002118:Calcium / Q000378:metabolism; D002121:Calcium Channel Blockers; D066298:In Vitro Techniques; D008564:Membrane Potentials / Q000187:drug effects; D009119:Muscle Contraction / Q000187:drug effects; D009131:Muscle, Smooth, Vascular / Q000187:drug effects* / Q000378:metabolism; D011169:Portal Vein / Q000187:drug effects; D051381:Rats; D013148:Spironolactone / Q000494:pharmacology*
D002121:Calcium Channel Blockers; D013148:Spironolactone; D002118:Calcium