Abstract:
AIMTo record L-type calcium current (
ICa,L) and T-type calcium current (
ICa,T) by perforated patch recording technique with β-escin in isolated guinea pig ventricular myocytes, and to investigate the effect of tetrandrine (Tet) on
ICa,L and
ICa,T. METHODSSingle myocytes were dissociated by enzymatic dissociation method.
ICa,L was recorded by perforated patch recording (PPR) and whole cell patch recording (WCR) techniques and
ICa,T was recorded by PPR. RESULTS25 μmol·L
-1 β-escin was shown to permeate the myocytes membrane and obtain PPR mode with success rate of 94% (16/17). Run-down of
ICa,L was considerably slower in PPR mode than in traditional WCR mode. Under PPR condition, 1~300 μmol·L
-1 Tet inhibited
ICa,L in a concentration-dependent manner with an IC
50 value of (15±5) μmol·L
-1 (95% confidence area: 10.6~18.9 μmol·L
-1,
N=6), 300 μmol·L
-1 Tet decreased
ICa,L from (-15.1±2.6) pA·pF
-1 to (-3.3±1.1) pA·pF
-1, with an inhibition rate of (77±10)%, (
P<0.01,
N=6). 300 μmol·L
-1 Tet shifted the current-voltage relation (
I~V) curves of
ICa,L upwardly, but it did not change the steady state activation curves of
ICa,L. The inhibition rate of 3, 30 and 300 μmol·L
-1 Tet on
ICa,T were (16±5)%, (40±7)% and (75±11)% (
P<0.01,
N=6) respectively. CONCLUSIONUsing 25 μmol·L
-1 β-escin can easily obtain stable PPR mode in isolated guinea pig ventricular myocytes. Under PPR condition, Tet was found to inhibit
ICa,L and
ICa,T concentration dependently.