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Probenecid inhibits adrenal zona glomerulosa cell aldosterone release
Author(s) -
Hanke Craig J.,
Danube Shana L.,
Hassemer Eryn,
Campbell William B.
Publication year - 2006
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.20.4.a345
Subject(s) - zona glomerulosa , aldosterone , probenecid , endocrinology , medicine , chemistry , pharmacology , receptor , angiotensin ii
This study examines the effect of the organic anion transporter inhibitor, probenecid, on aldosterone release and Fluo‐3 acetoxymethyl ester dye sequestration from cultured bovine adrenal zona glomerulosa (ZG) cells. ZG cells were isolated and maintained in primary culture in Hams F12 medium supplemented with antioxidants, fetal bovine serum and antibiotics prior to their use in aldosterone or Fluo‐3 experiments. Aldosterone release was quantified by enzyme‐linked immunosorbant assay. Probenecid inhibited angiotensin II‐stimulated aldosterone release in a concentration dependent manner and aldosterone release was completely inhibited with 10 mM probenecid. Potassium, adrenocorticotropic hormone, and 8‐bromoadenosine 3′,5′‐cyclic monophosphate‐stimulated aldosterone release was similarly inhibited by probenecid treatment. The inhibitory effects of probenecid were also observed in ZG cells stimulated with exogenous 25‐hydroxycholesterol and progesterone. Fluo‐3 fluorescence was increased by probenecid treatment. Treatment of ZG cells with 10 mM probenecid during the Fluo‐3 dye loading process caused a dramatic increase in visible fluorescence and more than doubled the average pixel intensity in fluorescing angiotensin II‐stimulated ZG cells as compared to control. We conclude that probenecid inhibits transport of Fluo‐3 dye by organic anion transporters in the cell membrane of the cultured ZG cell. We further conclude that probenecid interferes with either the enzymes involved in the process of aldosterone synthesis or a transporter required for aldosterone synthesis or release from the ZG cell. This work was supported by grants from the University of Wisconsin‐Green Bay.