Plasma marinobufagenin-like and ouabain-like immunoreactivity in adrenocorticotropin-treated rats

Recently, an endogenous digitalis-like factor (EDLF) was shown to be stimulated in corticotropin (ACTH) hypertension in the rat. We have shown that mammalian plasma contains a vasoconstrictor Na,K-ATPase inhibitor, which cross-reacts with an antibody to amphibian EDLF, marinobufagenin. In the present experiment, the effect of 8 days of intramuscular ACTH treatment (0.5 mg/kg/day) of male Fisher 344×NB rats on blood pressure, plasma ouabain-like and marinobufagenin-like immunoreactivity, and on the activity of Na,K-ATPase in aortic sarcolemma were studied. The ACTH treatment for 8 days resulted in increased systolic blood pressure (151 ± 12.4 v 121 ± 4.0 mm Hg, P< .01), inhibition of Na,K-ATPase in aortic sarcolemma (2.99 ± 0.35 v 5.43 ± 0.17 μmol ADP/mgprot/h), and increases in plasma concentration of marinobufagenin-like (0.44 ± 0.06 v 0.21 ± 0.05 nmol/L), but not ouabain-like (0.09 ± 0.01 v 0.10 ± 0.04 nmol/L) immunoreactivity. In dissociation enhanced lanthanide fluoroimmunoassay (DELFIA), serial dilutions of plasma from ACTH-treated rats extracted with 25% and 80% acetonitrile, respectively, demonstrated parallelism to the calibration curves of ouabain and marinobufagenin. These findings suggest that an endogenous bufodienolide Na,K-ATPase inhibitor, rather than an endogenous ouabain-like compound, is increased after 8 days of treatment of rats with ACTH.