Local anesthetics modulate neuronal calcium signaling through multiple sites of action

Abstract

Background: Local anesthetics (LAs) are known to inhibit voltage-dependent Na+ channels, as well as K+ and Ca2+ channels, but with lower potency. Since cellular excitability and responsiveness are largely determined by intracellular Ca2+ availability, sites along the Ca2+ signaling pathways may be targets of LAs. This study was aimed to investigate the LA effects on depolarization and receptor-mediated intracellular Ca2+ changes and to examine the role of Na+ and K+ channels in such functional responses. Methods: Effects of bupivacaine, ropivacaine, mepivacaine, and lidocaine (0.1-2.3 mM) on evoked [Ca2+]i transients were investigated in neuronal SH-SY5Y cell suspensions using Fura-2 as the intracellular Ca2+ indicator. Potassium chloride (KCl, 100 mM) and carbachol (1 mM) were individually or sequentially applied to evoke increases in intracellular Ca2+. Coapplication of LA and Na+/K+ channel blockers was used to evaluate the role of Na+ and K+ channels in the LA effect on the evoked [Ca2+]i transients. Results: All four LAs concentration-dependently inhibited both KCl- and carbachol-evoked [Ca2+]i transients with the potency order bupivacaine > ropivacaine > lidocaine ≥ mepivacaine. The carbachol-evoked [Ca2+]i transients were more sensitive to LAs without than with a KCl prestimulation, whereas the LA-effect on the KCl-evoked [Ca2+]i transients was not uniformly affected by a carbachol prestimulation. Na+ channel blockade did not alter the evoked [Ca2+]i transients with or without a LA. In the absence of LA, K+ channel blockade increased the KCl-, but decreased the carbachol-evoked [Ca2+]i transients. A coapplication of LA and K+ channel blocker resulted in larger inhibition of both KCl. and carbachol-evoked [Ca2+]i transients than by LA alone. Conclusions: Different and overlapping sites of action of LAs are involved in inhibiting the KCl- and carbachol-evoked [Ca2+]i transients, including voltage-dependent Ca2+ channels, a site associated with the caffeine-sensitive Ca2+ store and a possible site associated with the IP3-sensitive Ca2+ store, and a site in the muscarinic pathway. K+ channels, but not Na+ channels, seem to modulate the evoked [Ca2+]i transients, as well as the LA-effects on such responses.