Are Compounds Membrane-Associated or Present in the Cytosol? A Study Using Polyphenols in a Colon Carcinoma Cell Line Mo
- PDF / 516,830 Bytes
- 6 Pages / 595.276 x 790.866 pts Page_size
- 99 Downloads / 129 Views
REDOX MODULATORS (C JACOB, SECTION EDITOR)
Are Compounds Membrane-Associated or Present in the Cytosol? A Study Using Polyphenols in a Colon Carcinoma Cell Line Model Hannah Deusser 1 & Isabel Groh 2 & Tamara Bakuradze 1 & Nadine Simson 1 & Eva Kaiser 3 & Holger Barth 3 & Elke Richling 1 Accepted: 27 September 2020 # The Author(s) 2020
Abstract Purpose We investigated the cytosolic and membrane-associated contents of polyphenols after 4 hours of incubation (50 μM of each polyphenol) in the colon carcinoma cell line T84 using a novel, rapid, and convenient method based on permeabilization of the cell membrane using digitonin. The colon carcinoma cell line was used to investigate the intestinal uptake of polyphenols present in apple products. Recent Findings The results showed that hydroxycinnamic acids (caffeic and 5-caffeoylquinic acid) were only detected in the cytosolic fractions. In contrast, 0.3 to 8.2% of the initial concentrations (50 μM) of the flavonoids phloretin, quercetin, phloretin 2′-O-glucoside, and quercetin 3-O-rhamnoside were found in the membrane-associated fractions. In the cytosolic fractions, 0.2– 2.9% of these compounds were detected, corresponding to 25 to 40% of the total cell-associated (cytosolic plus membraneassociated fractions) polyphenol content. Summary Our results showed that after uptake, polyphenols were present in the cytosolic fraction of the cells as well as associated with the cell membrane. The presented method provides a useful in vitro tool for determining biologically active compounds in cellular fractions. Keywords Intracellular . Cytosol . Membrane . Digitonin . Polyphenols . Uptake . Colonic cells
Abbreviations DMEM DMSO EEA FCS Hsp90 HPLC-DAD LODs,
Dulbecco’s Modified Eagle’s Medium Dimethyl sulfoxide Early endosome antigen 1 Fetal calf serum Heat shock protein 90 High-performance liquid chromatography-diode array detection Limits of detection
This article is part of the Topical Collection on Redox Modulators * Elke Richling [email protected] 1
Department of Chemistry, Division of Food Chemistry and Toxicology, Technische Universität Kaiserslautern, Erwin-Schrödinger-Str. 52, 67663 Kaiserslautern, Germany
2
Institute of Experimental and Clinical Pharmacology and Toxicology, University of Tübingen, Wilhelmstraße 56, 72074 Tübingen, Germany
3
Institute of Pharmacology and Toxicology, University of Ulm Medical Center, Albert-Einstein-Allee 11, 89081 Ulm, Germany
LOQs SDS-PAGE S/N
Limits of quantitation Sodium dodecyl sulfate polyacrylamide gel electrophoresis Signal to noise
Introduction Polyphenols are secondary plant compounds frequently ingested in our daily diet. Fruits, vegetables, cereals, chocolate, and drinks (e.g., tea, wine, coffee, and fruit juices) are the major sources of polyphenols [1, 2]. Apples (Malus domestica Borkh.) contain between 5.2 and 27.2 g/kg dry weight polyphenols of various classes (hydroxycinnamic acids, dihydrochalcones, flavonols, and flavan-3-ols) [3]. Consuming apples can have several positive effects on human health,
Data Loading...
