Effects of Nitric Oxide on Voltage-Gated K(+) Currents in Human Cardiac Fibroblasts through the Protein Kinase G and Protein Kinase A Pathways but Not through S-Nitrosylation

This study investigated the expression of voltage-gated K(+) (K(V)) channels in human cardiac fibroblasts (HCFs), and the effect of nitric oxide (NO) on the K(V) currents, and the underlying phosphorylation mechanisms. In reverse transcription polymerase chain reaction, two types of K(V) channels were detected in HCFs: delayed rectifier K(+) channel and transient outward K(+) channel. In whole-cell patch-clamp technique, delayed rectifier K(+) current (I(K)) exhibited fast activation and slow inactivation, while transient outward K(+) current (I(to)) showed fast activation and inactivation kinetics. Both currents were blocked by 4-aminopyridine. An NO donor, S-nitroso-N-acetylpenicillamine (SNAP), increased the amplitude of I(K) in a concentration-dependent manner with an EC(50) value of 26.4 µM, but did not affect I(to). The stimulating effect of SNAP on I(K) was blocked by pretreatment with 1H-(1,2,4)oxadiazolo[4,3-a]quinoxalin-1-one (ODQ) or by KT5823. 8-bromo-cyclic GMP stimulated the I(K). The stimulating effect of SNAP on I(K) was also blocked by pretreatment with KT5720 or by SQ22536. Forskolin and 8-bromo-cyclic AMP each stimulated I(K). On the other hand, the stimulating effect of SNAP on I(K) was not blocked by pretreatment of N-ethylmaleimide or by DL-dithiothreitol. Our data suggest that NO enhances I(K), but not I(to), among K(V) currents of HCFs, and the stimulating effect of NO on I(K) is through the PKG and PKA pathways, not through S-nitrosylation.