Summary
Primary cultures of dermal fibroblasts from neonatal mice were used to investigate some of the anti-inflammatory effects of glucocorticoids in vitro as influenced by the genetic background of two different strains of mice (A/J and C57 B1/6J). Fibroblasts were cultured in the absence or presence of various glucocorticoids for 2–7 days. After 4–7 days in the presence of steroid, DNA synthesis was reduced by 50–85% while protein synthesis was inhibited by 50–60%. Corticosterone produced a dose-dependent inhibition of DNA synthesis in these cells with a 50% reduction occurring at 10 nM. Specific, high affinity, low capacity binding proteins for [3H]dexamethasone or [3H]triamcinolone acetonide were identified in the cytoplasm of neonatal dermal fibroblasts which had an apparent Kd of 9 nM and ∼5,200–6,400 binding sites/cell. Sedimentation analysis of the [3H]triamcinolone acetonide-receptor complexes on low salt glycerol gradients exhibited binding in the 7 to 8 S region of the gradients. These studies demonstrate that inhibition of growth of primary cultures of mouse neonatal dermal fibroblasts by glucocorticoids is probably mediated by a receptor-mediated pathway, and that this primary culture system might be useful in delineating other anti-inflammatory effects of glucocorticoids in vitro.
Zusammenfassung
Es wurden Primärkulturen dermaler Fibroblasten aus neugeborenen Mäusen verwandt, um einige der entzündungshemmenden Wirkungen der Glukocorticoide in vitro unter dem Einfluß des genetischen Hintergrunds zweier verschiedener Mäusestämme (A/J und C57B1/6J) zu untersuchen. Fibroblasten wurden 2-7 Tage lang mit und ohne verschiedene Glukocorticoide kultiviert. Unter Steroiden-4-7 Tage lang — wurde die DNA-Synthese um 50–85% und die Eiweißsynthese um 50–60% gehemmt. Corticosteron verursachte in diesen Zellen eine dosisabhängige Hemmung der DNA-Synthese, wobei eine 50%-Reduzierung bei 10 nM eintrat. Spezifisches Eiweiß mit hoher Affinität und niedriger Kapazität für [3H]-Dexamethasonbzw. [3H]-Triamzinolon-Acetonid wurde im Cytoplasma von Dermalfibroblasten offenbar mit einem Kd von 9 nM und ca. 5200–6400 Bindestellen/Zelle aufgefunden. Die Sedimentationsanalyse des [3H]-Triamzinolon-Acetonid-Receptorenkomplexes wies bei niedrigen Salzglyceringefällen eine Bindung in der 7-8S-Region auf. Diese Untersuchungen weisen darauf hin, daß die Wachstumshemmung von Primärkulturen dermaler Fibroblasten neugeborener Mäuse durch Glukocorticoide wahrscheinlich auf receptoren vermitteltem Wege stattfindet und daß dieses Primärkulturensystem zur Abgrenzung anderer entzündungshemmender Wirkungen, von Glukocorticoiden in vitro nützlich sein dürfte.
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This investigation was supported in part by a Public Health Service International Research Fellowship (FO-5-TWO 2236) to LAV
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Verbruggen, L.A., Salomon, D.S. Glucocorticoid receptors and inhibition of neonatal mouse dermal fibroblast growth in primary culture. Arch Dermatol Res 269, 111–126 (1980). https://doi.org/10.1007/BF00406531
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DOI: https://doi.org/10.1007/BF00406531