Mepivacaine reduces calcium transients in isolated murine ventricular cardiomyocytes

The potential mechanism of mepivacaine's myocardial depressant effect observed in papillary muscle has not yet been investigated at cellular level. Therefore, we evaluated mepivacaine's effects on Ca²⁺ transient in isolated adult mouse cardiomyocytes.

Single ventricular myocytes were enzymatically isolated from wild-type C57Bl/6 mice and loaded with 10 μM fluorescent Ca²⁺ indicator Fluo-4-AM to record intracellular Ca²⁺ transients upon electrical stimulation. The mepivacaine effects at half-maximal inhibitory concentration (IC₅₀) was determined on calibrated cardiomyocytes' Ca²⁺ transients by non-parametric statistical analyses on biophysical parameters. Combination of mepivacaine with NCX blockers ORM-10103 or NiCl₂ were used to test a possible mechanism to explain mepivacaine-induced Ca²⁺ transients' reduction.

A significant inhibition at mepivacaine's IC₅₀ (50 μM) on Ca²⁺ transients was measured in biophysical parameters such as peak (control: 528.6 ± 73.61 nM vs mepivacaine: 130.9 ± 15.63 nM; p < 0.05), peak area (control: 401.7 ± 63.09 nM*s vs mepivacaine: 72.14 ± 10.46 nM*s; p < 0.05), slope (control: 7699 ± 1110 nM/s vs mepivacaine: 1686 ± 226.6 nM/s; p < 0.05), time to peak (control: 107.9 ± 8.967 ms vs mepivacaine: 83.61 ± 7.650 ms; p < 0.05) and D₅₀ (control: 457.1 ± 47.16 ms vs mepivacaine: 284.5 ± 22.71 ms; p < 0.05). Combination of mepivacaine with NCX blockers ORM-10103 or NiCl₂ showed a significant increase in the baseline of [Ca²⁺] and arrhythmic activity upon electrical stimulation.

At cellular level, mepivacaine blocks Na⁺ channels, enhancing the reverse mode activity of NCX, leading to a significant reduction of Ca²⁺ transients. These results suggest a new mechanism for the mepivacaine-reduction contractility effect.