Effect of diethylstilbestrol (DES) on intracellular Ca 2 + levels in renal tubular cells

Add time:09/26/2019    Source:infona.pl

The effect of the estrogen diethylstilbestrol (DES) on intracellular Ca 2 + concentrations ([Ca 2 + ] i ) in Madin Darby canine kidney (MDCK) cells was investigated, using the fluorescent dye fura-2 as a Ca 2 + indicator. DES (10-50 μM) evoked [Ca 2 + ] i increases in a concentration-dependent manner. Extracellular Ca 2 + removal inhibited 45 +/- 5% of the Ca 2 + response. In Ca 2 + -free medium, pretreatment with 50 μM DES abolished the [Ca 2 + ] i increases induced by 2 μM carbonylcyanide m-chlorophenylhydrazone (CCCP; a mitochondrial uncoupler) and 1 μM thapsigargin (an endoplasmic reticulum Ca 2 + pump inhibitor); and pretreatment with CCCP and thapsigargin partly inhibited DES-induced [Ca 2 + ] i signals. Adding 3 mM Ca 2 + increased [Ca 2 + ] i in cells pretreated with 50 μM DES in Ca 2 + -free medium, suggesting that DES may induce capacitative Ca 2 + entry. 17β-Estradiol (2-20 μM) increased [Ca 2 + ] i , but 100 μM diethylstilbestrol dipropionate had no effect. Pretreatment with the phospholipase C inhibitor U73122 (1 μM) to abolish inositol 1,4,5-trisphosphate formation inhibited 30% of DES-induced Ca 2 + release. DES (20 μM) also increased [Ca 2 + ] i in human normal hepatocytes and osteosarcoma cells. Cumulatively, this study shows that DES induced rapid and sustained [Ca 2 + ] i increases by releasing intracellular Ca 2 + and triggering extracellular Ca 2 + entry in renal tubular cells.

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