Lowering Intracellular Calcium Concentration May Reduce the Cytotoxicity of Triamcinolone on Human Retinal Pigment Epithelial (ARPE19) Cells

C. Yeung, S. Chiang, K. Chan, C. Pang, D. Lam
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Abstract

1. The present study investigated the use of drugs that affect calcium (Ca2 +) levels and thus reduction of triamcinolone (TA)‐induced cytotoxicity on human retinal epithelial (ARPE19) cells. 2. Four groups were compared: ARPE19 cells alone, cells exposed to TA (0.1 mg/mL), cells that have been pretreated with one of the testing agents for 30 min before the addition of TA, and cells that have only been treated with one of the testing agents. Pinacidil (PIN) and its analogue, P1060, were used to test the effect of potassium (K+) channel opening on TA‐induced toxicity. Verapamil (VP) and diltiazem (DZ) were used to test their Ca2 + channel blocking effect. The cell viability under different settings was assessed using 3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide (MTT) assay. Ca2 +‐imaging was used to determine the changes in intracellular Ca2 + levels [(Ca2 +)i] upon different treatments. 3. Both PIN and P1060 reduced TA‐induced toxicity. Verapamil and DZ increased the viability of cells treated with TA significantly, suggesting that the excessive influx of Ca2 + was one of the main contributory factors to the TA‐induced toxicity. 4. The results suggest that the prevention of Ca2 + entry may be effective in the reduction of cell necrosis in the presence of TA.
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降低细胞内钙浓度可降低曲安奈德对人视网膜色素上皮细胞(ARPE19)的细胞毒性
1. 本研究调查了影响钙(Ca2 +)水平的药物的使用,从而降低曲安奈德(TA)诱导的对人视网膜上皮细胞(ARPE19)的细胞毒性。2. 比较四组:ARPE19细胞单独,暴露于TA (0.1 mg/mL)的细胞,在添加TA之前用一种测试剂预处理30分钟的细胞,以及仅用一种测试剂处理的细胞。Pinacidil (PIN)及其类似物P1060被用来测试钾离子通道打开对TA诱导的毒性的影响。用维拉帕米(VP)和地尔硫卓(DZ)检测它们对Ca2 +通道的阻断作用。采用3‐(4,5‐二甲基噻唑‐2‐基)‐2,5‐二苯基溴化四唑(MTT)测定法评估不同环境下的细胞活力。Ca2 +成像用于确定不同处理后细胞内Ca2 +水平[(Ca2 +)i]的变化。3.PIN和P1060均能降低TA诱导的毒性。维拉帕米和DZ显著提高了TA处理细胞的活力,这表明Ca2 +的过量流入是TA诱导毒性的主要因素之一。4. 结果表明,防止Ca2 +进入可能有效地减少TA存在下的细胞坏死。
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