Participation of cAMP in tacrine-induced gastric smooth muscle relaxation

Tacrine, a well-known acetylcholinesterase inhibitor, applied in concentrations higher than 2×10⁻⁵ mol/l promoted Ca²⁺-independent relaxation of rat gastric smooth muscles in experiments in vitro. The relaxation was not cholinergic and was a result of influence of tacrine over intracellular signaling pathways regulating smooth muscle contraction/relaxation. The nature of this untypical muscle relaxation was studied by using smooth muscle strips isolated from rat stomach. Their bioelectrical and mechanical responses were recorded after treatment with tacrine and different activators or blockers of intracellular pathways involved in muscle contractility. Following the activation of adenylate cyclase with 1×10⁻⁶ mol/l forskolin and increase in the concentration of cyclic adenosine monophosphate (cAMP) after application of 4×10⁻⁵ mol/l SQ22536, a significant decrease in the muscle relaxation was observed. Theophylline (2×10⁻⁴ mol/l), a phosphodiesterase inhibitor, had no effect on the amplitude of tacrine-induced relaxation. The latter was also reduced by inhibition of protein kinase A (PKA) with 5×10⁻⁶ mol/l KT5720. These findings support the assumption that tacrine promoted smooth muscle relaxation through PKA-induced phosphorylation and inhibition of myosin light chain kinase activity. The reduction of spike-linked Ca²⁺ influx provoked by tacrine was probably a secondary contributing process, associated with an influence of increased cAMP level on Ca²⁺ channels.