Author(s): Chang HT, Hsu SS, Chou CT, Cheng JS, Wang JL,
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Abstract AIMS: The effect of the natural product thymol on cytosolic Ca(2+) concentrations ([Ca(2+)](i)) and viability in MG63 human osteosarcoma cells was examined. METHODS: The Ca(2+)-sensitive fluorescent dye fura-2 was applied to measure [Ca(2+)](i). RESULTS: Thymol at concentrations of 200-1,000 μmol/l induced a [Ca(2+)](i) rise in a concentration-dependent fashion. The response was decreased partially by removal of extracellular Ca(2+). Thymol-induced Ca(2+) entry was inhibited by nifedipine, econazole, SK&F96365 and protein kinase C modulators. When extracellular Ca(2+) was removed, incubation with the endoplasmic reticulum Ca(2+) pump inhibitor thapsigargin or 2,5-di-tert-butylhydroquinone (BHQ) inhibited the thymol-induced [Ca(2+)](i) rise. Incubation with thymol also inhibited the thapsigargin or BHQ-induced [Ca(2+)](i) rise. Inhibition of phospholipase C with U73122 abolished the thymol-induced [Ca(2+)](i) rise. At concentrations of 100-600 μmol/l, thymol killed cells in a concentration-dependent manner. This cytotoxic effect was not changed by chelating cytosolic Ca(2+) with 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid/AM. Annexin V/propidium iodide staining data suggest that thymol (200 and 400 μmol/l) induced apoptosis in a concentration-dependent manner. Thymol (200 and 400 μmol/l) also increased levels of reactive oxygen species. CONCLUSIONS: In MG63 cells, thymol induced a [Ca(2+)](i) rise by inducing phospholipase C-dependent Ca(2+) release from the endoplasmic reticulum and Ca(2+) entry via protein kinase C-sensitive store-operated Ca(2+) channels. Thymol induced cell death that may involve apoptosis via mitochondrial pathways. Copyright © 2011 S. Karger AG, Basel.
This article was published in Pharmacology
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