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In vitro Corticosteroidbiosynthese in menschlichen Nebennieren

In vitro corticosteroid biosynthesis in human adrenals

I. Inkubation von Schnitten und Homogenaten frischen und tiefgefroren gelagerten Gewebes

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Summary

The formation of pregnenolone, progesterone, 11-desoxycorticosterone, corticosterone, aldosterone, 11-desoxycortisol and cortisol from endogenous precursors and the radioactivity incorporation into these steroids from added 4-14C-pregnenolone during in vitro incubation of normal human adrenal tissue were studied.

Results obtained from experiments with slices of fresh tissue were in good correlation with the secretory pattern of corticosteroids and the capacity of secretion in normal subjects. ACTH (“Synacthen” Ciba) stimulated the corticosteroid synthesis from endogenous precursors and had little effect on radioactivity incorporation.

Specific radioactivities of 11-desoxycorticosterone were 3–4 (slices of fresh tissue) and 12–14 (slices of stored tissue) times higher than those of progesterone indicating the possibility of an alternative pathway of corticosteroid synthesis in which progesterone is not an intermediate.

In slices of tissue stored at −20°C for 4–5 months ACTH had no effect on steroid formation, and there was a significant loss of 11β- and 17α-hydroxylation capacity as compared to fresh tissue slices.

The steroid pattern obtained from incubation of fresh tissue homogenates differed significantly from that obtained from corresponding experiments with slices and indicated that homogenization had caused defects in 11β- and 17α-hydroxylation. In contrast to the effects of ACTH on steroid synthesis in tissue slices, NADPH (NADP++G-6-P) in homogenates stimulated radioactivity incorporation to a higher degree than endogenous steroid formation. It caused a slight reactivation of 11β-hydroxylation which was more distinct for cortisol than for corticosterone.

Homogenates of stored tissue exhibited an additional loss of 11β- and 17α-hydroxylation capacity. There was no reactivation of 11β-hydroxylation by NADPH, but a slight reactivation of 17α-hydroxylation.

Zusammenfassung

Die Bildung von Pregnenolon, Progesteron, 11-Desoxycorticosteron, Corticosteron, Aldosteron, 11-Desoxycortisol und Cortisol aus endogenen Vorstufen und der Radioaktivitätseinbau in diese Steroide aus zugesetztem 4-14C-Pregnenolon während der in vitro Inkubation normalen menschlichen Nebennierengewebes wurden untersucht.

Ergebnisse aus Experimenten mit Schnitten frischen Gewebes zeigten gute Korrelation zum Corticosteroidsekretionsmuster und zur Sekretionskapazität bei Normalpersonen. ACTH („Synacthen“, Ciba) stimulierte die Corticosteroidsynthese aus endogenen Vorstufen und beeinflußte den Radioaktivitätseinbau nur geringfügig.

Die spezifischen Radioaktivitäten von 11-Desoxycorticosteron waren 3–4 (Schnitte frischen Gewebes) und 12–14 (Schnitte gelagerten Gewebes) mal höher als die des Progesterons. Daraus ist die mögliche Existenz eines alternativen Syntheseweges abzuleiten, in dem Progesteron als Zwischenstufe nicht vorkommt.

In Schnitten aus Gewebe, das 4–5 Monate bei −20°C gelagert worden war, zeigte ACTH keine Wirkung auf die Steroidbildung, und im Vergleich zu frischen Gewebeschnitten konnte ein signifikanter Kapazitätsverlust der 11β- und 17α-Hydroxylierung nachgewiesen werden.

Das aus der Inkubation von Homogenaten frischen Gewebes resultierende Steroidmuster unterschied sich signifikant von dem aus entsprechenden Experimenten mit Schnitten. Homogenisieren hatte offensichtlich zu Schädigungen im Bereich der 11β- und 17α-Hydroxylierung geführt. Im Gegensatz zur Wirkung des ACTH auf die Steroidsynthese in Gewebeschnitten stimulierte NADPH (NADP++G-6-P) in Homogenaten den Radioaktivitätseinbau stärker als die endogene Steroidbildung. In Homogenat frischen Gewebes bewirkte NADPH eine leichte Reaktivierung der 11β-Hydroxylierung, die für Cortisol stärker ausgeprägt war als für Corticosteron.

Homogenate aus gelagertem Gewebe zeigten zusätzliche Kapazitätsverluste der 11β- und 17α-Hydroxylierung. Unter der Einwirkung von NADPH ergab sich eine leichte Reaktivierung der 17α-Hydroxylierung, jedoch keine Reaktivierung der 11β-Hydroxylierung.

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Authors and Affiliations

  1. I. Medizinische Klinik und Poliklinik der Johannes Gutenberg-Universität, Mainz

    K. Sinterhauf, P. Herzog, G. Diedrichsen & D. Lommer

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  1. K. Sinterhauf
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  2. P. Herzog
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  3. G. Diedrichsen
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  4. D. Lommer
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  5. U. Aulbach
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Additional information

Herrn Professor Dr. H. P. Wolff zum 60. Geburtstag.

Die Arbeit wurde durch die Deutsche Forschungsgemeinschaft (Lo 118/7, SFB 36) und durch das Bundesministerium für Forschung und Technologie (St. Sch. 273) unterstützt.

Auszug aus der Dissertation von Peter Herzog, Fachbereich 08 Konservative Medizin der Johannes Gutenberg-Universität Mainz, 1974.

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Sinterhauf, K., Herzog, P., Diedrichsen, G. et al. In vitro Corticosteroidbiosynthese in menschlichen Nebennieren. Klin Wochenschr 52, 816–826 (1974). https://doi.org/10.1007/BF01468862

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