Abstract
Topical application of natural phenolic compound rosmarinic acid (ROSA) and its analogs is known to exert pharmacological effects, including anti-inflammation, antiallergy, and antioxidant properties. However, the mechanism of action of the ROSA remains largely unknown. Here, we describe a novel role of natural phenolic ROSA and its analogs in the selective inhibition of warmth-sensitive Ca(2+)-permeable cutaneous TRPV3 channel for the alleviation of skin lesions through the downregulation of NF-κB pathway. ROSA and its analogs, (E)-3-(3,4-dihydroxyphenyl)-N-(2-(3,4-dihydroxyphenyl) ethyl)-2-propenamide and methyl rosmarinate, inhibit macroscopic TRPV3 currents in both concentration-dependent and structure-dependent manners with IC(50) values ranging from 10 to 160 μM. ROSA also directly inhibits single TRPV3 channels by reducing the channel open probability without altering its unitary conductance. ROSA selectively targets TRPV3 over other subtypes of thermos-TRPs such as TRPV1, TRPV4, TRPA1 and TRPM8 channels. Site-directed mutagenesis combined with molecular docking reveals two residues T636 and T665 critical for ROSA-mediated inhibition of TRPV3. Furthermore, network pharmacology identified downstream p-P65, TNF-α, and interleukin-6 proteins in NF-κB signaling pathway as critically involved in ROSA-mediated reduction of cell death and alleviation of skin lesions. Altogether, our findings demonstrate that ROSA exerts its anti-inflammatory effects by selectively inhibiting TRPV3 channel and suppressing the NF-κB signaling pathway.