A cell-based assay system for activators of the environmental cell stress response.

Autor: Harbottle JA; Metabolic Health Group, Obesity & Metabolic Health Theme, Rowett Institute, UK., Petrie L; Metabolic Health Group, Obesity & Metabolic Health Theme, Rowett Institute, UK., Ruhe M; Metabolic Health Group, Obesity & Metabolic Health Theme, Rowett Institute, UK., Houssen WE; Marine Biodiscovery Centre, Chemistry Department, University of Aberdeen, Aberdeen, AB24 3UE, Scotland, UK; Institute of Medical Sciences, University of Aberdeen, Aberdeen, AB25 2ZD, Scotland, UK., Jaspars M; Marine Biodiscovery Centre, Chemistry Department, University of Aberdeen, Aberdeen, AB24 3UE, Scotland, UK., Kolb AF; Metabolic Health Group, Obesity & Metabolic Health Theme, Rowett Institute, UK. Electronic address: a.kolb@abdn.ac.uk.
Jazyk: angličtina
Zdroj: Analytical biochemistry [Anal Biochem] 2020 Mar 01; Vol. 592, pp. 113583. Date of Electronic Publication: 2020 Jan 13.
DOI: 10.1016/j.ab.2020.113583
Abstrakt: Improved health span and lifespan extension in a wide phylogenetic range of species is associated with the induction of the environmental cell stress response through a signalling pathway regulated by the transcription factor Nrf2. Phytochemicals which stimulate this response may form part of therapeutic interventions which stimulate endogenous cytoprotective mechanisms, thereby delaying the onset of age-related diseases and promoting healthy ageing in humans. In order to identify compounds that activate the Nrf2 pathway, a cell-based reporter system was established in HepG2 cells using a luciferase reporter gene under the control of the Nqo1 promoter. Sulforaphane, an isothiocyanate derived from cruciferous vegetables and a known activator of the Nrf2 pathway, was used to validate the reporter system. The transfected cell line HepG2 C1 was subsequently used to screen natural product libraries. Five compounds were identified as activating the bioluminescent reporter by greater than 5-fold. The two most potent compounds, MBC20 and MBC37, were further characterised and shown to stimulate endogenous cytoprotective gene and protein expression. The bioluminescent reporter system will allow rapid, in vitro identification of novel compounds that have the potential to improve health span through activation of the environmental stress response.
Competing Interests: Declaration of competing interest The authors declare that they have no conflict of interest.
(Copyright © 2020 Elsevier Inc. All rights reserved.)
Databáze: MEDLINE
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