OBJECTIVE: To investigate the role of adenosine tri-phosphate (ATP) purinergic signaling in mast cells (MCs) modulated by heat to further understand the molecular mechanisms of moxibustion. METHODS: Skin temperatures induced by monkshood cake moxibustion were evaluated by measuring the Neiguan acupoint (PC 6) from 31 participants with a digital thermocouple thermometer. Temperatures of 43 °C and 52 °C were applied to cultured human leukemia mast cell line HMC-1 in vitro. Calcium fluorescence was applied to detect intracellular Ca2+ ([Ca2+]). Extracellular ATP contents were measured by luciferin-luciferase assay. RESULTS: Maximum skin temperatures mostly ranged from 40-45 °C , but some reached up to 50 °C. Both 43 °C and 52 °C induced MC degranulation, which was accompanied by an increase in [Ca2+] and ATP release. Complexing extracellular Ca2+ with 5 mM ethylene glycol-bis (β-aminoethyl ether)-N,N,N',N'-tetraacetic acid (EGTA) inhibited the noxious, heat-induced elevation of [Ca2+]i and prevented the enhanced ATP secretion by those. cells at 52 °C, but not 43 °C. CONCLUSION: Monkshood cake moxibustion can generate heat sufficient to trigger cellular events of MCs, including degranulation, [Ca2+]i elevation, and ATP release, suggesting that purinergic signals originating from MCs are possibly the initiating response of acupoints to moxibustion.
OBJECTIVE: To investigate the role of adenosine tri-phosphate (ATP) purinergic signaling in mast cells (MCs) modulated by heat to further understand the molecular mechanisms of moxibustion. METHODS: Skin temperatures induced by monkshood cake moxibustion were evaluated by measuring the Neiguan acupoint (PC 6) from 31 participants with a digital thermocouple thermometer. Temperatures of 43 °C and 52 °C were applied to cultured humanleukemia mast cell line HMC-1 in vitro. Calcium fluorescence was applied to detect intracellular Ca2+ ([Ca2+]). Extracellular ATP contents were measured by luciferin-luciferase assay. RESULTS: Maximum skin temperatures mostly ranged from 40-45 °C , but some reached up to 50 °C. Both 43 °C and 52 °C induced MC degranulation, which was accompanied by an increase in [Ca2+] and ATP release. Complexing extracellular Ca2+ with 5 mM ethylene glycol-bis (β-aminoethyl ether)-N,N,N',N'-tetraacetic acid (EGTA) inhibited the noxious, heat-induced elevation of [Ca2+]i and prevented the enhanced ATP secretion by those. cells at 52 °C, but not 43 °C. CONCLUSION: Monkshood cake moxibustion can generate heat sufficient to trigger cellular events of MCs, including degranulation, [Ca2+]i elevation, and ATP release, suggesting that purinergic signals originating from MCs are possibly the initiating response of acupoints to moxibustion.