Effects of Thymol on Ca^(2+) Homeostasis and Apoptosis in MDCK Renal Tubular Cells

10.4077/CJP.2014.BAB192

Hong-Tai Chang；Chiang-Ting Chou；Wei-Zhe Liang；Ti Lu；Daih-Huang Kuo；Po-Chuen Shieh；Chin-Man Ho；Chung-Ren Jan

apoptosis ； Ca^(2+) ； MDCK ； thymol

The Chinese Journal of Physiology

57卷2期（2014 / 04 / 30）

90 - 98

英文

Thymol is a natural essential oil present in many plants and has many different effects in various cell types. However, the effect of thymol on the physiology of Madin-Darby canine kidney (MDCK) renal tubular cells is unknown. The action of the phytochemical thymol on cytosolic Ca^(2+) concentrations ([Ca^(2+)]i) and apoptosis in MDCK renal tubular cells was explored. Fura-2, a Ca^(2+)-sensitive fluorescent dye, was used to assess [Ca^(2+)]i. Thymol at concentrations of 200-500 μM caused a [Ca^(2+)]; rise in a concentration-dependent manner. Removal of extracellular Ca^(2+) partially reduced the effects of thymol. Thymol-induced Ca^(2+) entry was inhibited by nifedipine, econazole, SK&F96365 and protein kinase C modulators. In a Ca^(2+)-free medium, treatment with the endoplasmic reticulum Ca^(2+) pump inhibitor thapsigargin inhibited thymol-induced [Ca^(2+)]i increases. Treatment with thymol also inhibited thapsigargin-induced [Ca^(2+)]i rise. Thymol killed cells at concentrations of 300-500 μM in a concentration-dependent fashion. Chelating cytosolic Ca^(2+) with 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid/AM (BAPTA/AM) did not prevent thymol cytotoxicity. Thymol (400 and 500 μM) induced apoptosis detected by using Annexin V/propidium iodide staining. At 400 or 500 μM, thymol increased levels of reactive oxygen species (ROS). Together, in MDCK cells, thymol induced a [Ca^(2+)]i rise by inducing Ca^(2+) release from the endoplasmic reticulum and Ca^(2+) entry via protein kinase C-sensitive store-operated Ca^(2+) channels. Our data suggest that thymol-induced apoptosis might involve ROS production.

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