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Intracellular Ca 2+ mobilization is closely linked with the initiation of salivary secretion in parotid acinar cells. Reactive oxygen species (ROS) are known to be related to a variety of oxidative stress-induced cellular disorders and believed to be involved in salivary impairments. In this study, we investigated the underlying mechanism of hydrogen peroxide (H 2 O 2 ) on cytosolic Ca 2+ accumulation in mouse parotid acinar cells. Intracellular Ca 2+ levels were slowly elevated when 1 mM H 2 O 2 was perfused in the presence of normal extracellular Ca 2+ . In a Ca 2+ -free medium, 1 mM H 2 O 2 still enhanced the intracellular Ca 2+ level. Ca 2+ entry tested using manganese quenching technique was not affected by perfusion of 1 mM H 2 O 2 . On the other hand, 10 mM H 2 O 2 induced more rapid Ca 2+ accumulation and facilitated Ca 2+ entry from extracellular fluid. Ca 2+ refill into intracellular Ca 2+ store and inositol 1,4,5-trisphosphate (1 µM)-induced Ca 2+ release from Ca 2+ store was not affected by 1 mM H 2 O 2 in permeabilized cells. Ca 2+ efflux through plasma membrane Ca 2+ -ATPase (PMCA) was markedly blocked by 1 mM H 2 O 2 in thapsigargin-treated intact acinar cells. Antioxidants, either catalase or dithiothreitol, completely protected H 2 O 2 -induced Ca 2+ accumulation through PMCA inactivation. From the above results, we suggest that excessive production of H 2 O 2 under pathological conditions may lead to cytosolic Ca 2+ accumulation and that the primary mechanism of H 2 O 2 -induced Ca 2+ accumulation is likely to inhibit Ca 2+ efflux through PMCA rather than mobilize Ca 2+ ions from extracellular medium or intracellular stores in mouse parotid acinar cells.

Hydrogen peroxide inhibits Ca2+ efflux through plasma membrane Ca2+-ATPase in mouse parotid acinar cells

Hydrogen peroxide inhibits Ca²⁺ efflux through plasma membrane Ca²⁺-ATPase in mouse parotid acinar cells