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Hydrogen peroxide inhibits Ca 2+ efflux through plasma membrane Ca 2+ -ATPase in mouse parotid acinar cells.

Intracellular Ca2+ 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 (H2 O2 ) on cytosolic Ca2+ accumulation in mouse parotid acinar cells. Intracellular Ca2+ levels were slowly elevated when 1 mM H2 O2 was perfused in the presence of normal extracellular Ca2+ . In a Ca2+ -free medium, 1 mM H2 O2 still enhanced the intracellular Ca2+ level. Ca2+ entry tested using manganese quenching technique was not affected by perfusion of 1 mM H2 O2 . On the other hand, 10 mM H2 O2 induced more rapid Ca2+ accumulation and facilitated Ca2+ entry from extracellular fluid. Ca2+ refill into intracellular Ca2+ store and inositol 1,4,5-trisphosphate (1 µM)-induced Ca2+ release from Ca2+ store was not affected by 1 mM H2 O2 in permeabilized cells. Ca2+ efflux through plasma membrane Ca2+ -ATPase (PMCA) was markedly blocked by 1 mM H2 O2 in thapsigargin-treated intact acinar cells. Antioxidants, either catalase or dithiothreitol, completely protected H2 O2 -induced Ca2+ accumulation through PMCA inactivation. From the above results, we suggest that excessive production of H2 O2 under pathological conditions may lead to cytosolic Ca2+ accumulation and that the primary mechanism of H2 O2 -induced Ca2+ accumulation is likely to inhibit Ca2+ efflux through PMCA rather than mobilize Ca2+ ions from extracellular medium or intracellular stores in mouse parotid acinar cells.

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