Contributions of nitric oxide, EDHF, and EETS to endothelium-dependent relaxation in renal afferent arterioles

Dan Wang, Lillian J. Borrego-Conde, John R. Falck, Kamalesh K. Sharma, Christopher S. Wilcox, Jason G. Umans

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Abstract

Background. Acetylcholine-induced endothelium-dependent relaxation in the renal afferent arteriole has been ascribed to nitric oxide, but the role of endothelium-derived hyperpolarizing factors (EDHFs) and 14,15-epoxyeicosatrienoic acid (14,15-EET) are unclear. Methods. Single afferent arterioles were dissected from kidney of normal rabbits and microperfused in vitro at 60 mm Hg. Vessels were preconstricted submaximally with norepinephrine (10-8 mol/L). Relaxation was assessed following cumulative addition of ACh (10-9 to 10-4 mol/L) alone, or in the presence of indomethacin (to inhibit cyclooxygenase), NW-nitro-L-arginine (L-NNA) (to inhibit nitric oxide synthase), methylene blue (to inhibit soluble guanylate cyclase), or a combination of L-NNA + methylene blue. To assess contributions by EDHF, studies were repeated with either apamin + charybdotoxin [to block Ca2+-activated K+ channels (KCa)] or with 40 mmol/L KCl. To asses the role of 14,15-EET, relaxations were evaluated in the presence of its competitive inhibitor 14,15-epoxyeicosa-5(Z)-enoic acid (14,15-EEZE). Results. Relaxation by acetylcholine was abolished following endothelial denudation. It was unaffected by indomethacin but was inhibited 54% ± 5% (P < 0.001) by L-NNA, 57% ± 5% by methylene blue, and 60% ± 4% by the combination of L-NNA plus methylene blue. Relaxation was inhibited further by KCl (80% ± 6%) or by apamin + charybdotoxin (96% ± 2%). 14,15-EEZE, alone, inhibited acetylcholine-induced relaxation by 29% ± 3%, and by 80% ± 5% in the presence of L-NNA. Conclusion. Acetylcholine-induced afferent arteriolar relaxation depends strongly on both nitric oxice, acting via soluble guanylate cyclase, and on an EDHF, likely 14,15-EET, acting via KCa.

Original languageEnglish (US)
Pages (from-to)2187-2193
Number of pages7
JournalKidney International
Volume63
Issue number6
DOIs
Publication statusPublished - Jun 1 2003

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Keywords

  • 14,15-epoxyeicosa-5(Z)-enoic acid
  • Calcium-activated potassium channel
  • Endothelium-dependent hyperpolarizing factor
  • Endothelium-derived relaxing factor
  • Epoxyeicosatrienoic acids
  • Soluble guanylyl cyclase

ASJC Scopus subject areas

  • Nephrology

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