Untersuchungen zur Regulation von hCG durch Kortisol (Prednisolon) in der Trophoblastzelle in vitro

Zusammenfassung

Fragestellung: Die Trophoblastzelle synthetisiert eine Reihe von Hormonen, wobei die hCG-Synthese eine herausragende Rolle spielt. Durch ihre besondere enzymatische Ausstattung kann sie die Reaktion Kortisol ↔ Kortison katalysieren. In-vitro-Stimulierungsversuche an isolierten Trophoblastzellen mit CRH, ACTH und Prednisolon zeigten, dass in Trophoblastzellen ein dem Hypophyse/Hypothalamus entsprechender Regelkreislauf existiert. In dieser Studie beschreiben wir den Einfluss von Kortisol (Prednisolon) auf die hCG-Produktion von Trophoblastzellen. Methode und Methodik: Die Trophoblastzellisolierung erfolgt nach grober Präparation des Plazentagewebes mittels mehrschrittiger DNAse I und Trypsinverdauung. Die gewonnene Zellsuspension wird auf die definierte Zellkonzentration von 1 × 10⁶ Zellen/ml eingestellt und in Nährmedium kultiviert. Nach jeweils 8 Stunden erfolgt die Zugabe von Prednisolon. Nach jeweils 24 Stunden wird die hCG-Konzentration gemessen. Parallel werden die hCG-Werte einer nichtstimulierten Kultur bestimmt. Ergebnisse: Die Zugabe von Prednisolon (50 µg/ml) stimuliert die hCG-Konzentration zeitabhängig. Schlussfolgerung: Die Trophoblastzelle weist in vitro eine erhöhte hCG-Sekretion unter Prednisolonstimulierung auf. Erstmals konnte eine hCG-Stimulierung mit Prednisolon innerhalb der Trophoblastzelle in vitro demonstriert werden.

Abstract

Objective: Trophoblast cells synthesize a variety of hormones, in which hCG plays a major role. On the strength of special enzymes they are capable of catalyzing the reaction cortisol ↔ cortisone. In vitro experiments showed the influence on ACTH- and cortisol secretion by CRH, ACTH and prednisolon. In this study we describe the influence of cortisol (prednisolon) on hCG production of trophoblast cells in vitro. Material and methods: Trophoblast cells were prepared from human term placentae by standard trypsin-DNAse dispersion of villous tissue followed by a percoll gradient centrifugation step. After adjusting the cell suspension to a defined cell concentration of 1 × 10⁶ cells/ml cells were cultivated. The addition of prednisolon followed every eight hours. The samples were collected after 24 hours for a total of 96 hours also from unstimulated cultures. Culture supernatants were assayed for hCG by enzyme-immunometric methods. Results: The addition of prednisolon (50 µg/ml) stimulates the concentration of hCG in a time-depending manner. Conclusions: The trophoblast cell shows an increase in the concentration of hCG after stimulation with cortisol. For the first time an influence of cortisol (prednisolon) on hCG production could be demonstrated in cultured trophoblast cells.

Schlüsselwörter

Trophoblastzelle - hCG - Kortisol - in vitro

Key words

Trophoblast cells - hCG - cortisol - in vitro

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