Effect of Nortriptyline on Intracellular Ca(superscript 2+) Handling and Viability in Canine Renal Tubular Cells

He-Hsiung Cheng；Chun-Jen Huang；Wei-Chuan Chen；I-Shu Chen；Shiuh-Inn Liu；Shu-Shong Hsu；Hong-Tai Chang；Jong-Khing Wang；Yih-Chau Lu；Chung-Ren Jan

Ca(superscript 2+) signaling ； fura-2 ； MDCK cells ； nortriptyline ； thapsigargin

The Chinese Journal of Physiology

49卷6期（2006 / 12 / 01）

283 - 289

英文

In Madin-Darby canine kidney (MDCK) cells, the effect of nortriptyline, an antidepressant, on intracellular Ca(superscript 2+) concentration ([Ca(superscript 2+)](subscript i)) was measured by using fura-2. Nortriptyline (＞10 μM) caused a rapid increase of [Ca(superscript 2+)](subscript i) in a concentration-dependent manner (EC50 = 75 μM). Nortriptyline-induced [Ca(superscript 2+)](subscript i) increase was prevented by 40% by removal of extracellular Ca(superscript 2+) but was not altered by voltagegated Ca(superscript 2+) channel blockers. In Ca(superscript 2+)-free medium, thapsigargin, an inhibitor of the endoplasmic reticulum Ca(superscript 2+)-ATPase, caused a monophasic [Ca(superscript 2+)](subscript i) increase, after which the increasing effect of nortriptyline on [Ca(superscript 2+)](subscript i) was abolished; also, pretreatment with nortriptyline reduced a large portion of thapsigargin-induced [Ca(superscript 2+)](subscript i) increase. U73122, an inhibitor of phospholipase C, abolished ATP (but not nortriptyline)-induced [Ca(superscript 2+)](subscript i) increase. Overnight incubation with 10 μM nortriptyline decreased cell viability by 16%, and 50 μM nortriptyline killed all cells. Prechelation of cytosolic Ca(superscript 2+) with BAPTA did not alter nortriptyline-induced cell death. These findings suggest that nortriptyline rapidly increased [Ca(superscript 2+)](subscript i) in renal tubular cells by stimulating both extracellular Ca(superscript 2+) influx and intracellular Ca(superscript 2+) release, and was cytotoxic at higher concentrations in a Ca(superscript 2+)-dissociated manner.

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