二吲哚甲烷对 MDCK 肾小管细胞内钙离子稳态和活力的影响。
Effect of diindolylmethane on Ca(2+) homeostasis and viability in MDCK renal tubular cells.
机构信息
Department of Laboratory Medicine, Zuoying Armed Forces General Hospital, Kaohsiung, Taiwan.
出版信息
Hum Exp Toxicol. 2013 Apr;32(4):344-53. doi: 10.1177/0960327112462727.
The effect of the natural product diindolylmethane (DIM) on cytosolic Ca(2+) concentrations ([Ca(2+)]i) and viability in MDCK renal tubular cells was explored. The Ca(2+)-sensitive fluorescent dye fura-2 was applied to measure [Ca(2+)]i. DIM at concentrations 1-50 μM induced a [Ca(2+)]i rise in a concentration-dependent manner. The response was reduced partly by removing Ca(2+). DIM induced Mn(2+) influx leading to quenching of fura-2 fluorescence. DIM-evoked Ca(2+) entry was suppressed by nifedipine, econazole, SK&F96365 and protein kinase C modulators. In the absence of extracellular Ca(2+), incubation with the endoplasmic reticulum Ca(2+) pump inhibitor thapsigargin (TG) or 2,5-di-tert-butylhydroquinone (BHQ) greatly inhibited DIM-induced [Ca(2+)]i rise. Incubation with DIM abolished TG or BHQ-induced [Ca(2+)]i rise. Inhibition of phospholipase C with U73122 reduced DIM-induced [Ca(2+)]i rise by 50%. At 1, 10, 40 and 50 μM, DIM slightly enhanced cell proliferation. The effect of 50 μM DIM was reversed by chelating cytosolic Ca(2+) with 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid. In sum, in MDCK cells, DIM induced a [Ca(2+)]i rise by evoking phospholipase C-dependent Ca(2+) release from the endoplasmic reticulum and Ca(2+) entry via protein kinase C-sensitive store-operated Ca(2+) channels. DIM did not induce cell death.
研究了天然产物二吲哚甲烷（DIM）对 MDCK 肾小管细胞胞浆游离 Ca²⁺浓度([Ca²⁺]i)和活力的影响。应用 Ca²⁺敏感荧光染料 fura-2 测量[Ca²⁺]i。1-50 μM 的 DIM 以浓度依赖的方式诱导 [Ca²⁺]i 升高。部分去除 Ca²⁺可减少反应。DIM 诱导 Mn²⁺内流，导致 fura-2 荧光猝灭。硝苯地平、依康唑、SK&F96365 和蛋白激酶 C 调节剂抑制 DIM 诱导的 Ca²⁺内流。在不存在细胞外 Ca²⁺的情况下，用内质网 Ca²⁺泵抑制剂 thapsigargin (TG)或 2,5-二叔丁基对苯二酚 (BHQ)孵育可显著抑制 DIM 诱导的 [Ca²⁺]i 升高。用 DIM 孵育可消除 TG 或 BHQ 诱导的 [Ca²⁺]i 升高。用 U73122 抑制磷脂酶 C 可使 DIM 诱导的 [Ca²⁺]i 升高减少 50%。在 1、10、40 和 50 μM 时，DIM 轻微增强细胞增殖。50 μM DIM 的作用可通过螯合胞浆 Ca²⁺用 1,2-双(2-氨基苯氧基)乙烷-N,N,N',N'-四乙酸来逆转。总之，在 MDCK 细胞中，DIM 通过激活磷脂酶 C 从内质网释放 Ca²⁺依赖性 Ca²⁺释放和蛋白激酶 C 敏感的储存操纵性 Ca²⁺通道引起 Ca²⁺内流，从而诱导 [Ca²⁺]i 升高。DIM 不会诱导细胞死亡。
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