Novel inhibitory effects of cardamonin on thromboxane A2-induced scratching response: Blocking of Gh/transglutaminase-2 binding to thromboxane A2 receptor.

Alpinia katsumadai is known to suppress thromboxane A2 (TXA2) receptor agonist-induced scratching in mice. The specific components of A. katsumadai responsible for these biological effects, however, are not known. In the present study, we investigated whether cardamonin (CDN), one of major principles of A. katsumadai, has suppressive effects on TXA2-induced scratching in mice. Scratching induced by U46619 (the TXA2 receptor agonist) at a dose of 10nmol/site was shown to be suppressed by CDN (0.1nmol-0.5nmol/site). Suppression of the U46619-induced scratching response by CDN was found to be unrelated to competition with the ligand at the TXA2 receptor, since CDN did not suppress [(3)H] SQ29548 (the TXA2 receptor antagonist) binding to TXA2 receptor. TXA2 receptor expression in A549, HaCaT, and SH-SY5Y cell lines was examined and determined to be significant in the A549 and SH-SY5Y cell lines. Further, binding of high molecular G protein Gh/transglutaminase-2 (Gh/Tgase-2) to TXA2 receptor was confirmed in the A549 and SH-SY5Y cells by co-immunoprecipitation. CDN suppressed the binding of TXA2 receptor with Gh/Tgase-2, which also acts as a G protein involved in TXA2 signaling. These results suggested that CDN suppresses TXA2 receptor agonist-induced scratching by suppressing TXA2 signaling, specifically via blocking of the binding of Gh/Tgase-2 to TXA2 receptor.