Uptake and metabolism of 12-hydroxyeicosatetraenoic acid (12-HETE) by cultured renal tubular epithelial cells (RTEC)

Conference · Sat Mar 01 04:00:00 EST 1986 · Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States)

OSTI ID:5384694

Gordon, J A; Spector, A A

To determine if 12-HETE, a lipoxygenase product that mediates inflammation and tissue injury, can interact with RTEC, confluent Madin Darby Canine Kidney (MDCK) cells were incubated for 2-16 hr with 1.0 ..mu..M (/sup 3/H)-12-HETE. Initial uptake of 12-HETE was rapid; at 16 hrs. 70% of the 12-HETE uptake was incorporated into phospholipids (PL). The distribution among the choline, ethanolamine, inositol, and serine PL was 36, 36, 20 and 8%, respectively. Incubation of MDCK cells with 0.5 to 5.0 ..mu..M (/sup 3/H)-12-HETE for 1 hr indicated linear uptake without evidence of saturation. Incubation with 1.0 ..mu..M 12-HETE and 0.25-10.0 ..mu..M arachidonic acid for 1 hr revealed no competition for uptake at the lower concentrations but a 40% reduction in 12-HETE uptake at 10.0 ..mu..M. Polarity of 12-HETE uptake was indicated by a preference of the basolateral surface over the apical surface by 1.4. After 2 hr, analysis of the medium by reverse phase HPLC revealed that 12-HETE was converted to three polar metabolites which eluted at 25.9, 29.4 and 31.3 min respectively; 12-HETE eluted at 37.5 min. The appearance of these polar metabolites was not prevented by ibuprofen (50 ..mu..M) nordihydroguaiaretic acid (30 ..mu..M), allopurinol (15 mM), or butylated hydroxytoluene (20 ..mu..M). These findings suggest that the lipoxygenase product 12-HETE may affect RTEC through incorporation into membrane PL and/or conversion to polar metabolites.

• Research Organization: Univ. of Iowa College of Medicine, Iowa City
• OSTI ID: 5384694
• Report Number(s): CONF-8604222-
• Conference Information: Journal Name: Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States) Journal Volume: 45:3
• Country of Publication: United States
• Language: English

Related Subjects

550501* -- Metabolism-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
ANIMAL TISSUES
BODY
CARBOXYLIC ACIDS
CELL CONSTITUENTS
CELL CULTURES
CELL MEMBRANES
CHROMATOGRAPHY
EICOSANOIC ACID
EPITHELIUM
ESTERS
ISOTOPE APPLICATIONS
KIDNEYS
LABELLED COMPOUNDS
LIPIDS
LIQUID COLUMN CHROMATOGRAPHY
MEMBRANES
METABOLISM
METABOLITES
MONOCARBOXYLIC ACIDS
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANIC PHOSPHORUS COMPOUNDS
ORGANS
PHOSPHOLIPIDS
SEPARATION PROCESSES
TISSUES
TRACER TECHNIQUES
TRITIUM COMPOUNDS
UPTAKE