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Caffeine-induced block of Na+ current in guinea pig single ventricular cells
Y Habuchi1, H Tanaka, T Furukawa
1Department of Laboratory Medicine, Kyoto Prefectural University of Medicine, Japan.
Abstract:
Effects of caffeine on Na+ current (INa) were investigated in single ventricular cells from guinea pigs using the whole cell clamp method. With a Ca(2+)-containing internal solution (pCa 8.2), 10 mM caffeine blocked INa by 17.5 +/- 4.6% at a -120-mV holding potential. It was accompanied by 3- to 5-mV shifts of the steady-state inactivation curve and time constant-voltage relationship toward hyperpolarization. The inactivation kinetics spontaneously shifted toward hyperpolarization at 0.30 +/- 0.17 mV/min. The spontaneous shift was accompanied by a similar negative shift of the threshold potential, whereas caffeine did not affect it. Caffeine retarded the recovery of INa from inactivation, and a 4-mV positive shift in the recovery potential produced a similar retardation in INa recovery without caffeine. The INa block by caffeine was not influenced by reinforcing the internal buffering capacities using internal solutions containing 40 mM ethylene glycol-bis(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid or 50 mM N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid or by pretreating the cell with 1 microM ryanodine. Neither pretreatment with isoproterenol or H 7 nor prestimulation of Gs protein by nonhydrolyzable GTP (GTP gamma S) altered the effects of caffeine on INa. It is concluded that caffeine inhibits INa and shifts the inactivation kinetics without being mediated by changes in intracellular ionic composition or intracellular signaling systems. Direct action on the channel proteins may be involved.
Insights
Caffeine inhibits sodium current (INa) in heart cells, altering inactivation properties. This effect appears to be a direct action on channel proteins, independent of intracellular signaling pathways.
Area of Science:
- Cardiology
- Electrophysiology
- Pharmacology
Background:
- Sodium current (INa) is crucial for cardiac action potential generation.
- Caffeine is a widely consumed stimulant with known physiological effects.
Purpose of the Study:
- To investigate the direct effects of caffeine on cardiac INa in guinea pig ventricular cells.
- To elucidate the mechanisms underlying caffeine's influence on INa kinetics and inactivation.
Main Methods:
- Whole-cell patch clamp technique was employed on isolated guinea pig ventricular myocytes.
- Experiments utilized a calcium-containing internal solution (pCa 8.2).
- Caffeine's effects on INa, inactivation curves, and recovery kinetics were analyzed.
Main Results:
- Caffeine (10 mM) significantly blocked INa by 17.5%.
- Caffeine induced hyperpolarizing shifts in steady-state inactivation and recovery from inactivation.
- Caffeine slowed the recovery of INa from inactivation, suggesting altered channel gating.
Conclusions:
- Caffeine directly inhibits cardiac INa and modifies its inactivation properties.
- These effects are independent of intracellular ion concentrations and common signaling pathways.
- Direct interaction of caffeine with cardiac sodium channel proteins is proposed.