Curcumin ((E,E)-1,7-bis(4-hydroxy-3-methoxyphenyl)-1,6-heptadiene-3,5-dione) activates and desensitizes the nociceptor ion channel TRPA1

Abstract

The ion channel TRPA1 is activated by a wide variety of noxious stimuli, such as pollutants, products of oxidative tissue damage, and pungent natural products. Many TRPA1 activators are reactive electrophiles that form Michael adducts with cysteine and lysine residues of TRPA1's intracellular N-terminus. Curcumin, the active principle of turmeric root (Curcuma longa), can also form Michael adducts. In order to test the hypothesis that the electrophilic curcumin activates TRPA1, we have performed whole-cell, voltage-clamp analysis on both HEK293 cells expressing human TRPA1 (hTRPA1-HEK) and native mouse vagal neurons. In nominally calcium-free extracellular and intracellular solutions which minimized the chances of calcium-dependent activation of TRPA1, curcumin increased TRPA1 currents in hTRPA1-HEK cells in a concentration-dependent manner (1-30μM) but did not cause block or activation of recombinant TRPM8 and TRPV1. In addition, 7 out of 11 vagal sensory neurons from wild type mice responded to curcumin (30μM) with inward currents (11.6±5.4pA/pF) that were largely reversed by TRPA1 blockers. In marked contrast, neurons from TRPA1-deficient mice did not respond to curcumin (30μM). With physiological levels of calcium added to the external solution to facilitate channel desensitization, curcumin-dependent currents in hTRPA1-HEK cells were completely desensitized and exhibited marked tachyphylaxis upon subsequent application of curcumin. Taken together, these results demonstrate that curcumin causes activation and subsequent desensitization of native and recombinant TRPA1 ion channels of multiple mammalian species.