Measurement of endogenous Na+,K+‐ATPase inhibitors in human plasma and urine using high‐performance liquid chromatography

FEBS Letters

Crabos, M.¹  Cloix, J.F.¹  Wainer, I.W.² 

[1] U7 INSERM, Department of Pharmacology, Hôpital Necker, 161 rue de Sévres, F-75015 Paris, France;Division of Drug Chemistry, U.S. Food and Drug Administration, Washington, DC 20204, USA

关键词: (Na++K+)-ATPase;    Endogenous (Na+ + K+)-ATPase inhibitor;    Plasma inhibitor;    Urine inhibitor;    Reverse-phase HPLC;   

DOI  :  10.1016/0014-5793(84)80946-1

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

This study was undertaken to assess endogenous Na⁺,K⁺-ATPase inhibitors in both plasma and urine in the same subjects. Samples were chromatographed on reverse-phase HPLC using an acetonitrile gradient and the eluent screened using Na⁺,K⁺-ATPase inhibition and cross-reaction with anti-digoxin antibodies. The donors were divided into inhibiting and non-inhibiting subjects using a previously described method, plasma action on ouabain binding and on Na⁺,K⁺-ATPase activity. Three Na⁺,K⁺-ATPase inhibitors (IP, 2P and 3P) were detectable in plasma; the antibodies cross-reaction of the peaks 2P and 3P were larger than that of peak IP. The peaks 2P and 3P were significantly higher in inhibiting subjects as compared to non-inhibiting subjects. The 24-h urine is resolved into two peaks inhibiting Na⁺,K⁺-ATPase activity (1U and 2U). Peak 2U cross-reacted with anti-digoxin antibodies to a greater extent than peak 1U and is significantly larger in inhibiting subjects in terms of Na⁺,K⁺-ATPase inhibition. These data support the heterogeneity of human Na⁺,K⁺-ATPase inhibitor in both plasma and urine.

【 授权许可】

Unknown