Summary
1. The intraperitoneal assay of insulin activity has been used to detect the presence of an insulin inhibitor in serum. 2. A solution of 5% bovine serum albumin has been established as an injection medium permitting the maximum insulin effectiveness. 3. When insulin was injected intraperitoneally in serum from male rats or male human subjects, the insulin effectiveness on the diaphragm was reduced compared with that of insulin in 5% bovine albumin; whereas that on the epididymal adipose tissue was unaffected. 4. The inhibitory activity was associated with the albumin fraction of the serum; and neither the inhibitory activity of the serum nor that of the albumin derived from the serum was affected by prolonged dialysis. 5. The inhibitory activity was released from the albumin by boiling at pH 4 to 5. The inhibitory substance appears to be a polypeptide of mol. wt between 5000 and 10000.
Résumé
1. Le dosage biologique de l'activité insulinique selon la technique de l'injection intrapéritonéale a été utilisé pour détecter l'existence d'un inhibiteur sérique de l'insuline. 2. Il a été établi que la solution d'injection permettant d'obtenir un effet insulinique maximum est celle qui contient de l'albumine bovine, à raison de 5 g pour cent. 3. On constate que l'insuline mélangée, soit à du sérum humain normal, soit à du sérum de rats mâles normaux, est moins efficace dans son action sur le diaphragme que l'insuline injectée seule dans la solution d'albumine bovine à 5%. L'effet sur le tissu adipeux par contre demeure inchangé. 4. L'activité inhibitrice ainsi observée est associée à la fraction albumine du sérum. Elle ne disparaît ni après dialyse prolongée du sérum complet ni après celle de la fraction albumine seule. 5. L'activité inhibitrice est libérée de l'albumine quand on fait bouillir celle-ci, le pH étant de 4 à 5. Il semble que la substance obtenue de cette manière soit un polypeptide d'un poids moléculaire de 5000 à 10000.
Zusammenfassung
1. Die Messung der Wirkung intraperitoneal injizierten Insulins am Zwerchfell und am epididymalen Fettgewebe erlaubt den Nachweis von Inhibitoren der Insulinaktivität in der Injektionsflüssigkeit. 2. In einer 5%igen Lösung von Rinder-Albumin hat intraperitoneal injiziertes Insulin einen maximalen Effekt. 3. Die Insulinwirkung am Zwerchfell, nicht aber am Fettgewebe, wird signifikant vermindert, wenn das Hormon zusammen mit Seren von männlichen Ratten und Menschen injiziert wird. 4. Die Inhibitor-Aktivität befindet sich in der Albuminfraktion der betreffenden Seren und wird durch Dialyse des Serums oder der Albuminfraktion nicht vermindert. 5. Sie kann nach Sieden bei pH 4 bis 5 vom Albumin abgetrennt werden. Bei der inhibitorisch aktiven Substanz scheint es sich um ein Peptid mit MG zwischen 5000 und 10000 zu handeln.
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References
Bray, G.A.: A simple efficient liquid scintillator for counting aqueous solutions in a liquid scintillation counter. Analyt. Biochem. 1, 279–286 (1960).
Debro, J.R., H. Tarver and A. Korner: The determination of serum albumin and globulin by a new method. J. Lab. clin. Med. 50, 728–732 (1957).
Dole, V.P., and H. Meinertz: Microdetermination of long-chain fatty acids in plasma and tissues. J. biol. Chem. 235, 2595–2599 (1960).
Ensinck, J.W., P.L. Poffenbarger, R.A. Hogan and R.H. Williams: An artifact in the measurement of the Synalbumin insulin antagonist in serum. Abstr. 26th Ann. Meeting, Amer. Diabetes Assoc. Diabetes 15, 514 (1966).
Jervell, J.: The antagonistic effect of human plasma albumin on the insulin-stimulated glucose uptake of the isolated rat diaphragm. Acta physiol. scand. 65, 33–44 (1965).
Lilley, M.D., and J. Vallance-Owen: A factor associated with “Visking” tubing capable of antagonizing insulin. Nature (Lond.) 190, 1196–1197 (1961).
Rafaelsen, O.J.: Glycogen content of rat diaphragm after intraperitoneal injection of insulin and other hormones. Acta physiol. scand. 61, 314–322 (1964).
—: Insulin-like activity of human serum determined by glycogen increase of diaphragm after intraperitoneal injection into the intact rat. Acta physiol. scand. 61, 323–330 (1964).
—, V. Lauris and A.E. Renold: Localized intraperitoneal action of insulin on rat diaphragm and epididymal adipose tissue in vivo. Diabetes 14, 19–26 (1965).
Randle, P.J., P.B. Garland, C.N. Hales and S.A. Newsholme: The glucose-fatty acid cycle and the metabolic disturbances of diabetes mellitus. Lancet 1963 I, 785–789.
Richter, J.: A new method for the fractionation of serum proteins. Acta chim. Acad. Sci. hung. 41, 167–169 (1964).
Stauffacher, W., O.B. Crofford, B. Jeanrenaud and A.E. Renold: Comparative studies of muscle and adipose tissue metabolism in lean and obese mice. Ann. N.Y. Acad. Sci. 131, 528–540 (1965).
Worthing, A.G., and J. Geffner: Treatment of experimental data. p. 461. New York: J. Wiley and Sons 1959.
Young, D.A.B.: The penetration of sugars into the cells of animal tissues: the action of insulin on the efflux of galactose from the isolated perfused rat heart. D. Phil, thesis. University of Oxford (1960).
—: Effect of change in length of day on the insulin sensitivity of the rat heart. Nature (Lond.) 193, 488 (1962).
—: Hypothalamic (photoperiodic) control of a seasonal antagonism to insulin in the rat heart. J. Physiol. (Lond.) 178, 530–543 (1965).
-, and B. Benson: A serum inhibitor of insulin action on muscle. 2. Assessment of inhibitory activity (to be published).
-,- A serum inhibitor of insulin action on muscle. 3. Pituitary-dependence and other endocrine controls (to be published).
Young, F.G.: Insulin and insulin antagonism. Endocrinology 73, 654–664 (1963).
References
Feinstein, R.N.: An apparatus for increasing the speed of dialysis of small quantities of fluids. J. Lab. clin. Med. 40, 313–315 (1952).
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Supported by the Fonds National Suisse de la Recherche Scientifique, Grant No. 3618.
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Young, D.A.B. A serum inhibitor of insulin action on muscle. Diabetologia 3, 287–298 (1967). https://doi.org/10.1007/BF00429860
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DOI: https://doi.org/10.1007/BF00429860