Cheng H-H, Chou C-T, Sun T-K, Liang W-Z, Cheng J-S, Chang H-T, Tseng H-W, Kuo C-C, Chen F-A, Kuo D-H, Shieh P, Jan C-R
Department of Medicine, Chang Bing Show Chwan Memorial Hospital, Changhua County, Taiwan.
Department of Nursing, Division of Basic Medical Sciences, Chang Gung Institute of Technology, Chia-Yi, Taiwan Chronic Diseases and Health Promotion Research Center, Chang Gung Institute of Technology, Chia-Yi, Taiwan.
Hum Exp Toxicol. 2015 Nov;34(11):1096-105. doi: 10.1177/0960327115569810. Epub 2015 Jan 30.
Naproxen is an anti-inflammatory drug that affects cellular calcium ion (Ca(2+)) homeostasis and viability in different cells. This study explored the effect of naproxen on Ca(2+) and viability in Madin-Darby canine kidney cells (MDCK) canine renal tubular cells. At concentrations between 50 μM and 300 μM, naproxen induced Ca(2+) rises in a concentration-dependent manner. This Ca(2+) signal was reduced partly when extracellular Ca(2+) was removed. The Ca(2+) signal was inhibited by a Ca(2+) channel blocker nifedipine but not by store-operated Ca(2+) channel inhibitors (econazole and SKF96365), a protein kinase C (PKC) activator phorbol 12-myristate 13-acetate, and a PKC inhibitor GF109203X. In Ca(2+)-free medium, pretreatment with 2,5-di-tert-butylhydroquinone or thapsigargin, an inhibitor of endoplasmic reticulum Ca(2+) pumps, partly inhibited naproxen-induced Ca(2+) signal. Inhibition of phospholipase C with U73122 did not alter naproxen-evoked Ca(2+) rises. At concentrations between 15 μM and 30 μM, naproxen killed cells in a concentration-dependent manner, which was not reversed by prechelating cytosolic Ca(2+) with the acetoxymethyl ester of 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid acetoxymethyl. Annexin V/propidium iodide staining data suggest that naproxen induced apoptosis. Together, in MDCK renal tubular cells, naproxen induced Ca(2+) rises by inducing Ca(2+) release from multiple stores that included the endoplasmic reticulum and Ca(2+) entry via nifedipine-sensitive Ca(2+) channels. Naproxen induced cell death that involved apoptosis.
萘普生是一种抗炎药物，可影响不同细胞中的细胞钙离子（Ca(2+)）稳态和活力。本研究探讨了萘普生对犬肾近曲小管细胞（MDCK）中[Ca(2+)]i和活力的影响。在50μM至300μM的浓度范围内，萘普生以浓度依赖性方式诱导[Ca(2+)]i升高。当去除细胞外Ca(2+)时，这种Ca(2+)信号部分降低。Ca(2+)信号被Ca(2+)通道阻滞剂硝苯地平抑制，但不受储存-操作性Ca(2+)通道抑制剂（酮康唑和SKF96365）、蛋白激酶C（PKC）激活剂佛波醇12-肉豆蔻酸酯13-乙酸酯和PKC抑制剂GF109203X的抑制。在无Ca(2+)培养基中，用2,5-二叔丁基对苯二酚或毒胡萝卜素（内质网Ca(2+)泵抑制剂）预处理可部分抑制萘普生诱导的Ca(2+)信号。用U73122抑制磷脂酶C不会改变萘普生诱发的[Ca(2+)]i升高。在15μM至30μM的浓度范围内，萘普生以浓度依赖性方式杀死细胞，用1,2-双（2-氨基苯氧基）乙烷-N,N,N',N'-四乙酸乙酰氧甲酯预螯合胞质Ca(2+)并不能逆转这种作用。膜联蛋白V/碘化丙啶染色数据表明萘普生诱导细胞凋亡。总之，在MDCK肾小管细胞中，萘普生通过诱导包括内质网在内的多个储存库释放Ca(2+)以及通过硝苯地平敏感的Ca(2+)通道使Ca(2+)内流，从而诱导[Ca(2+)]i升高。萘普生诱导的细胞死亡涉及细胞凋亡。
