# Direct Effects of Capsaicin on Voltage-Dependent Calcium Channels of Mammalian Skeletal Muscle

## TL;DR

This study shows that capsaicin and its analogs inhibit calcium channels in skeletal muscle, independent of TRPV1 receptors.

## Contribution

The novel finding is that capsaicin inhibits L-type calcium channels in skeletal muscle without involving TRPV1 receptors.

## Key findings

- Capsaicin inhibited depolarization-induced Ca2+ effluxes in rabbit skeletal muscle membranes.
- Capsaicinoids with hydrocarbon chains inhibited Ca2+ effluxes, while vanilloids did not.
- Molecular simulations showed capsaicinoids bind to CaV1.1 channels similarly to amlodipine.

## Abstract

Capsaicin, a naturally occurring polyphenol, is known to affect energy expenditure and muscle fatigue and modulate contractions in skeletal muscle. The L-type Ca2+ channels are known to be an important ion channel involved in the various muscle functions and the effect of capsaicin on the skeletal L-type Ca2+ channels is currently unknown. In this study, the effects of capsaicin and capsaicin analogs on depolarization-induced Ca2+ effluxes through L-type Ca2+ channels in transverse tubule membranes from rabbit skeletal muscle and L-type Ca2+ currents recorded using the whole-cell patch clamp technique in rat myotubes were examined. Capsaicin, in the concentration range of 3–100 µM, inhibited depolarization-induced Ca2+ effluxes. The effect of capsaicin was not reversed by TRPV1 antagonist SB-366791 (10 µM). While vanilloids (30 µM) including vanillin, vanillyl alcohol, and vanillylamine were ineffective, other capsaicinoids (30 µM) including dihydrocapsaicin, nonivamide, and nordihydrocapsaicin significantly inhibited Ca2+ effluxes, suggesting that hydrocarbon chains are required for inhibition. In rat myotubes, capsaicin inhibited L-type Ca2+ currents with an IC50 value of 27.2 μM in the presence of SB-366791. Furthermore, in docking studies and molecular dynamic simulations, capsaicinoids with an aliphatic tail showed stronger binding and stable bent conformations in CaV1.1, forming hydrogen bonds with Ser1011 and Thr935 and hydrophobic/π–alkyl contacts with Phe1008, Ile1052, Met1366, and Ala1369, resembling the binding mode of amlodipine. In conclusion, the results indicate that the function of L-type Ca2+ channels in mammalian skeletal muscle was inhibited by capsaicin and capsaicin analogs in a TRPV1-independent manner.