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Increasing evidence has revealed that glibenclamide has a wide range of anti-inflammatory effects. However, it is unclear whether glibenclamide can affect the resting and adenosine triphosphate (ATP)-induced intracellular calcium ([Ca 2+ ] i ) handling in Raw 264.7 macrophages. In the present study, [Ca 2+ ] i  transient, reactive oxygen species (ROS) and mitochondrial activity were measured by the high-speed TILLvisION digital imaging system using the indicators of Fura 2-am, DCFDA and rhodamine-123, respectively. We found that glibenclamide, pinacidil and other unselective K +  channel blockers had no effect on the resting [Ca 2+ ] i  of Raw 264.7 cells. Extracellular ATP (100 µM) induced [Ca 2+ ] i  transient elevation independent of extracellular Ca 2+ . The transient elevation was inhibited by an ROS scavenger (tiron) and mitochondria inhibitor (rotenone). Glibenclamide and 5-hydroxydecanoate (5-HD) also decreased ATP-induced [Ca 2+ ] i  transient elevation, but pinacidil and other unselective K +  channel blockers had no effect. Glibenclamide also decreased the peak of [Ca 2+ ] i  transient induced by extracellular thapsigargin (Tg, 1 µM). Furthermore, glibenclamide decreased intracellular ROS and mitochondrial activity. When pretreated with tiron and rotenone, glibenclamide could not decrease ATP, and Tg induced maximal [Ca 2+ ] i  transient further. We conclude that glibenclamide may inhibit ATP-induced [Ca 2+ ] i  transient elevation by blocking mitochondria K ATP  channels, resulting in decreased ROS generation and mitochondrial activity in Raw 264.7 macrophages.

plos one

Glibenclamide Decreases ATP-Induced Intracellular Calcium Transient Elevation via Inhibiting Reactive Oxygen Species and Mitochondrial Activity in Macrophages