Tyrosine kinase inhibitor, genistein, inhibits macroscopic L-type calcium current in rat portal vein smooth muscle cells.
The effect of genistein, a specific tyrosine kinase inhibitor, was tested on the slow (L-type) Ca2+ current (ICa(L)) of vascular smooth muscle cells from freshly isolated rat portal vein, using whole-cell voltage clamp. To isolate ICa(L), the pipette contained high Cs+ and the bath contained 140 mM tetraethylammonium (TEA) to block K+ currents. Bath application of genistein decreased ICa(L) in a concentration-dependent manner within 3-6 min. The concentration for half-maximal inhibition (IC50) was 54.9 microM (at a holding potential of -40 mV). At a concentration of 300 microM, genistein produced nearly complete inhibition of ICa(L). The inhibitory effect of genistein was not reversed after washout for up to 5 min. The potential for half-inhibition (V1/2) of the steady-state inactivation curve for ICa(L) was shifted to the left by genistein (10.6 mV at 50 microM), suggesting that genistein exerts a voltage-dependent block. Superfusion with daidzein, an inactive analog of genistein, had no inhibitory effect on ICa(L) at concentrations as high as 300 microM. These results may suggest that the L-type Ca2+ channels in vascular smooth muscle cells are possibly modulated by endogenous tyrosine kinase activity. That is, tonic phosphorylation by tyrosine kinases maintains the Ca2+ channels in an available state for activation by depolarization. Thus, the vascular tone may be controlled by tyrosine kinase activity.
Liu
H
H
Department of Anesthesiology, University of Texas-Houston Medical School 77030, USA.
Li
K
K
Sperelakis
N
N
D000818:Animals; D002121:Calcium Channel Blockers / Q000494:pharmacology*; D015220:Calcium Channels / Q000187:drug effects* / Q000502:physiology; D020746:Calcium Channels, L-Type; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D019833:Genistein / Q000494:pharmacology*; D066298:In Vitro Techniques; D007529:Isoflavones / Q000494:pharmacology; D008297:Male; D008564:Membrane Potentials / Q000187:drug effects; D009131:Muscle, Smooth, Vascular / Q000166:cytology / Q000187:drug effects* / Q000201:enzymology / Q000502:physiology; D018408:Patch-Clamp Techniques; D011169:Portal Vein / Q000187:drug effects* / Q000201:enzymology / Q000502:physiology; D011505:Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors*; D051381:Rats; D017207:Rats, Sprague-Dawley
D002121:Calcium Channel Blockers; D015220:Calcium Channels; D020746:Calcium Channels, L-Type; D004791:Enzyme Inhibitors; D007529:Isoflavones; C004742:daidzein; D019833:Genistein; D011505:Protein-Tyrosine Kinases