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AJP - Cell Physiology, Vol 266, Issue 6 C1775-C1782, Copyright © 1994 by American Physiological Society
ARTICLES |
B. Escalante, D. Erlij, J. R. Falck and J. C. McGiff
Department of Pharmacology, New York Medical College, Valhalla 10595.
The medullary thick ascending limb of Henle's loop (mTALH) of the rabbit metabolizes arachidonic acid (AA) via a cytochrome P-450 (P-450) monooxygenase pathway to several products, of which the principal are 20-hydroxy-5,8,11,14-eicosatetraenoic acid (20-HETE) and 1,20-eicosatetraenedioic acid (20-COOH-AA). To understand their mechanism of action on alkali cation metabolism in mTALH cells, we have compared their effects with those of ouabain and furosemide. Incubation of rabbit isolated mTALH cells with either 1 mM ouabain or furosemide decreased K+ content from a control of 1,015 +/- 51 peq/micrograms protein to 717 +/- 41 and to 548 +/- 48 peq/micrograms protein, respectively, whereas they had opposite effects on Na+ content; from a control of 138 +/- 22 peq/micrograms protein, ouabain increased Na+ content to 357 +/- 37 peq/micrograms protein, and furosemide decreased it to 64 +/- 23 peq/micrograms protein. Preincubation with either 20-HETE (1 microM) or 20-COOH-AA (1 microM) decreased Na+ and K+, resembling furosemide in their effects on Na+ and K+ content. In other experiments we used monensin-treated cells to determine 86Rb uptake under conditions in which Na+ entry into the cell was not rate limiting. Under these conditions ouabain still inhibited 86Rb uptake, and the effect of AA was blocked. A major action of AA metabolites on Na(+)-K(+)-adenosinetriphosphatase was thereby excluded. Furthermore, AA metabolites did not inhibit Ba(2+)-sensitive 86Rb efflux, indicating that they do not act through K+ channels.(ABSTRACT TRUNCATED AT 250 WORDS)
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