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Simultaneous measurement of endothelium‐derived relaxing factor by bioassay and guanylate cyclase stimulation
Author(s) -
Kondo Kigen,
Mitchell Jane Allison,
Nucci Gilberto,
Vane John Robert
Publication year - 1989
Publication title -
british journal of pharmacology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.432
H-Index - 211
eISSN - 1476-5381
pISSN - 0007-1188
DOI - 10.1111/j.1476-5381.1989.tb12637.x
Subject(s) - endothelium derived relaxing factor , stimulation , bradykinin , chemistry , sodium nitroprusside , nitric oxide , radioimmunoassay , medicine , gucy1a3 , endocrinology , cyclic guanosine monophosphate , gucy1b3 , superoxide dismutase , guanylate cyclase 2c , cyclase , biochemistry , biology , enzyme , receptor
1 Endothelium‐derived relaxing factor (EDRF) released by cultured endothelial cells (EC) from bovine aortae was measured by bioassay using pre‐contracted strips of rabbit aorta and by radioimmunoassay of guanosine 3′:5′‐cyclic monophosphate (cyclic GMP) produced by stimulation of bovine lung soluble guanylate cyclase. 2 Bradykinin (Bk, 3 and 30 pmol) injected through a column of EC caused release of EDRF as detected by bioassay and increased cyclic GMP concentrations. Superoxide dismutase (SOD, 15 u ml −1 ) increased the amount of EDRF detected by the activation of soluble guanylate cyclase. 3 In the absence of endothelial cells, nitric oxide (NO, 1–2 μ m ), arachidonic acid (AA, 3–30 μ m ) or sodium nitröprusside (SNP, 1–100 μ m ) stimulated guanylate cyclase. Superoxide dismutase strongly increased the stimulation of guanylate cyclase induced by NO, but had little effect on the stimulation induced by SNP and no effect on the stimulation induced by AA. 4 Oxyhaemoglobin (10–300 μ m ) abolished the stimulation of guanylate cyclase by EDRF, NO or SNP but was much less effective as an inhibitor of AA‐induced stimulation of guanylate cyclase. 5 These results demonstrate that measurement of guanylate cyclase stimulation by radioimmunoassay is a viable method for detecting EDRF release, especially useful when the drugs used interfere with bioassay tissues.