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Monitoring cytochrome P450 activity in living hepatocytes by chromogenic substrates in response to drug treatment or during cell maturation.
Theobald, Jannick; Cheng, Xinlai; Ghanem, Ali; Gaitantzi, Haristi; Song, Guangqi; Klipp, Edda; Wodke, Judith; Becker, Holger; Mrowka, Ralf; Breitkopf-Heinlein, Katja; Dooley, Steven; Wölfl, Stefan.
Afiliação
  • Theobald J; Institute of Pharmacy and Molecular Biotechnology, Pharmaceutical Biology, Heidelberg University, Im Neuenheimer Feld 364, 69120, Heidelberg, Germany.
  • Cheng X; Institute of Pharmacy and Molecular Biotechnology, Pharmaceutical Biology, Heidelberg University, Im Neuenheimer Feld 364, 69120, Heidelberg, Germany.
  • Ghanem A; Institute of Pharmacy and Molecular Biotechnology, Pharmaceutical Biology, Heidelberg University, Im Neuenheimer Feld 364, 69120, Heidelberg, Germany.
  • Gaitantzi H; Department of Medicine II, Section Molecular Hepatology, Alcohol Associated Diseases, Medical Faculty Mannheim at Heidelberg University, Theodor-Kutzer-Ufer 1-3, 68167, Mannheim, Germany.
  • Song G; Department of Gastroenterology, Shanghai Institute of Liver Disease, Zhongshan Hospital, Fudan University, Building 19, Fenglin Road 179, Shanghai, China.
  • Klipp E; Theoretical Biophysics, Department of Biology, Humboldt-Universität zu Berlin, Invalidenstraße 42, 10115, Berlin, Germany.
  • Wodke J; Theoretical Biophysics, Department of Biology, Humboldt-Universität zu Berlin, Invalidenstraße 42, 10115, Berlin, Germany.
  • Becker H; microfluidic ChipShop GmbH, Stockholmer Str. 20, 07747, Jena, Germany.
  • Mrowka R; Experimentelle Nephrologie, KIM III, Universitätsklinikum Jena, Am Nonnenplan 2, 07743, Jena, Germany.
  • Breitkopf-Heinlein K; Department of Medicine II, Section Molecular Hepatology, Alcohol Associated Diseases, Medical Faculty Mannheim at Heidelberg University, Theodor-Kutzer-Ufer 1-3, 68167, Mannheim, Germany.
  • Dooley S; Department of Medicine II, Section Molecular Hepatology, Alcohol Associated Diseases, Medical Faculty Mannheim at Heidelberg University, Theodor-Kutzer-Ufer 1-3, 68167, Mannheim, Germany.
  • Wölfl S; Institute of Pharmacy and Molecular Biotechnology, Pharmaceutical Biology, Heidelberg University, Im Neuenheimer Feld 364, 69120, Heidelberg, Germany. wolfl@uni-hd.de.
Arch Toxicol ; 92(3): 1133-1149, 2018 Mar.
Article em En | MEDLINE | ID: mdl-29209748
The metabolic activity of hepatocytes is a central prerequisite for drug activity and a key element in drug-drug interaction. This central role in metabolism largely depends on the activity of the cytochrome P450 (CYP450) enzyme family, which is not only dependent on liver cell maturation but is also controlled in response to drug and chemical exposure. Here, we report the use of VividDye fluorogenic CYP450 substrates to directly measure and continuously monitor metabolic activity in living hepatocytes. We observed time- and dose-dependent correlation in response to established and putative CYP450 inducers acting through the aryl hydrocarbon receptor and drug combinations. Using repetitive addition of VividDye fluorogenic substrate on a daily basis, we demonstrated the new application of VividDye for monitoring the maturation and dedifferentiation of hepatic cells. Despite a lack of high specificity for individual CYP450 isoenzymes, our approach enables continuous monitoring of metabolic activity in living cells with no need to disrupt cultivation. Our assay can be integrated in in vitro liver-mimetic models for on-line monitoring and thus should enhance the reliability of these tissue model systems.
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Bioensaio / Compostos Cromogênicos / Hepatócitos / Sistema Enzimático do Citocromo P-450 Tipo de estudo: Prognostic_studies Limite: Animals / Humans / Male Idioma: En Revista: Arch Toxicol Ano de publicação: 2018 Tipo de documento: Article País de afiliação: Alemanha

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Bioensaio / Compostos Cromogênicos / Hepatócitos / Sistema Enzimático do Citocromo P-450 Tipo de estudo: Prognostic_studies Limite: Animals / Humans / Male Idioma: En Revista: Arch Toxicol Ano de publicação: 2018 Tipo de documento: Article País de afiliação: Alemanha