Mechanical and electrophysiological effects of 8-oxoberberine (JKL1073A) on atrial tissue

The effects of 8-oxoberberine (JKL1073A) on contractions and electrophysiological characteristics of atrial tissues were examined. In driven left atria of the rat JKL1073A (10-100 microM) increased twitch tension dose-dependently. In spontaneously beating right atria, JKL1073A increased twitch tension but decreased beating rate slightly. The positive inotropic and the negative chronotropic effect of 30 microM JKL1073A was not affected by prazosin (1 microM), propranolol (1 microM) and 3-isobutyl-1-methyl-xanthine (10 microM) but significantly suppressed by 4-aminopyridine (2 mM 4-AP). Current-clamp study revealed that JKL1073A prolonged rat atrial action potential duration (APD). This prolongation of APD by JKL1073A was decreased by pretreating the cells with 2 mM 4-AP. Voltage-clamp study showed that JKL1073A inhibited the integral of the transient outward current (I(to)) dose-dependently with a KD value of 3.66 +/- 0.93 microM in rat atrial myocytes. The equilibrium dissociation constant (Kd) for JKL1073A bindings to open state I(to) was 0.50 +/- 0.08 microM. The suppression of I(to) by 3 microM JKL1073A was accompanied by shortening of its inactivation time constant from 52.5 +/- 0.9 ms to 16.8 +/- 0.7 ms. V(0.5) for the steady-state inactivation curve of I(to) was shifted from -25.7 +/- 3.3 mV to -34.8 +/- 3.2 mV. In human atrial cells, similar inhibition of I(to) and prolongation of APD by JKL1073A was found. The KD value of JKL1073A for inhibition of the integral of I(to) in human atrial cells is 4.03 +/- 0.02 microM. The Kd for bindings to open state I(to) is 0.5 microM. Currents through K1 channels of rat and human atrial myocytes were not inhibited by JKL1073A at concentrations up to 10 microM. These results indicate that JKL1073A exerts a positive inotropic effect by inhibition of I(to). JKL1073A inhibit I(to) by binding to open state channels or shifting of the steady-state inactivation curve of I(to).