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Protein Profiling of a Cellular Model of NAFLD by Advanced Bioanalytical Approaches
Advanced quantitative bioanalytical approaches in combination with network analyses allow us to answer complex biological questions, such as the description of changes in protein profiles under disease conditions or upon treatment with drugs. In the present work, three quantitative proteomic approac...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9408868/ https://www.ncbi.nlm.nih.gov/pubmed/36012291 http://dx.doi.org/10.3390/ijms23169025 |
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author | Altomare, Alessandra Anna Aiello, Gilda Garcia, Jessica Leite Garrone, Giulia Zoanni, Beatrice Carini, Marina Aldini, Giancarlo D’Amato, Alfonsina |
author_facet | Altomare, Alessandra Anna Aiello, Gilda Garcia, Jessica Leite Garrone, Giulia Zoanni, Beatrice Carini, Marina Aldini, Giancarlo D’Amato, Alfonsina |
author_sort | Altomare, Alessandra Anna |
collection | PubMed |
description | Advanced quantitative bioanalytical approaches in combination with network analyses allow us to answer complex biological questions, such as the description of changes in protein profiles under disease conditions or upon treatment with drugs. In the present work, three quantitative proteomic approaches—either based on labelling or not—in combination with network analyses were applied to a new in vitro cellular model of nonalcoholic fatty liver disease (NAFLD) for the first time. This disease is characterized by the accumulation of lipids, inflammation, fibrosis, and insulin resistance. Hepatic G2 cells were used as model, and NAFLD was induced by a complex of oleic acid and bovine albumin. The development of the disease was verified by lipid vesicle staining and by the increase in the expression of perilipin-2—a protein constitutively present in the vesicles during NAFLD. The nLC–MS/MS analyses of peptide samples obtained from three different proteomic approaches resulted in accurate and reproducible quantitative data of protein fold-change expressed in NAFLD versus control cells. The differentially regulated proteins were used to evaluate the involved and statistically enriched pathways. Network analyses highlighted several functional and disease modules affected by NAFLD, such as inflammation, oxidative stress defense, cell proliferation, and ferroptosis. Each quantitative approach allowed the identification of similar modulated pathways. The combination of the three approaches improved the power of statistical network analyses by increasing the number of involved proteins and their fold-change. In conclusion, the application of advanced bioanalytical approaches in combination with pathway analyses allows the in-depth and accurate description of the protein profile of an in vitro cellular model of NAFLD by using high-resolution quantitative mass spectrometry data. This model could be extremely useful in the discovery of new drugs to modulate the equilibrium NAFLD health state. |
format | Online Article Text |
id | pubmed-9408868 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-94088682022-08-26 Protein Profiling of a Cellular Model of NAFLD by Advanced Bioanalytical Approaches Altomare, Alessandra Anna Aiello, Gilda Garcia, Jessica Leite Garrone, Giulia Zoanni, Beatrice Carini, Marina Aldini, Giancarlo D’Amato, Alfonsina Int J Mol Sci Article Advanced quantitative bioanalytical approaches in combination with network analyses allow us to answer complex biological questions, such as the description of changes in protein profiles under disease conditions or upon treatment with drugs. In the present work, three quantitative proteomic approaches—either based on labelling or not—in combination with network analyses were applied to a new in vitro cellular model of nonalcoholic fatty liver disease (NAFLD) for the first time. This disease is characterized by the accumulation of lipids, inflammation, fibrosis, and insulin resistance. Hepatic G2 cells were used as model, and NAFLD was induced by a complex of oleic acid and bovine albumin. The development of the disease was verified by lipid vesicle staining and by the increase in the expression of perilipin-2—a protein constitutively present in the vesicles during NAFLD. The nLC–MS/MS analyses of peptide samples obtained from three different proteomic approaches resulted in accurate and reproducible quantitative data of protein fold-change expressed in NAFLD versus control cells. The differentially regulated proteins were used to evaluate the involved and statistically enriched pathways. Network analyses highlighted several functional and disease modules affected by NAFLD, such as inflammation, oxidative stress defense, cell proliferation, and ferroptosis. Each quantitative approach allowed the identification of similar modulated pathways. The combination of the three approaches improved the power of statistical network analyses by increasing the number of involved proteins and their fold-change. In conclusion, the application of advanced bioanalytical approaches in combination with pathway analyses allows the in-depth and accurate description of the protein profile of an in vitro cellular model of NAFLD by using high-resolution quantitative mass spectrometry data. This model could be extremely useful in the discovery of new drugs to modulate the equilibrium NAFLD health state. MDPI 2022-08-12 /pmc/articles/PMC9408868/ /pubmed/36012291 http://dx.doi.org/10.3390/ijms23169025 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Altomare, Alessandra Anna Aiello, Gilda Garcia, Jessica Leite Garrone, Giulia Zoanni, Beatrice Carini, Marina Aldini, Giancarlo D’Amato, Alfonsina Protein Profiling of a Cellular Model of NAFLD by Advanced Bioanalytical Approaches |
title | Protein Profiling of a Cellular Model of NAFLD by Advanced Bioanalytical Approaches |
title_full | Protein Profiling of a Cellular Model of NAFLD by Advanced Bioanalytical Approaches |
title_fullStr | Protein Profiling of a Cellular Model of NAFLD by Advanced Bioanalytical Approaches |
title_full_unstemmed | Protein Profiling of a Cellular Model of NAFLD by Advanced Bioanalytical Approaches |
title_short | Protein Profiling of a Cellular Model of NAFLD by Advanced Bioanalytical Approaches |
title_sort | protein profiling of a cellular model of nafld by advanced bioanalytical approaches |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9408868/ https://www.ncbi.nlm.nih.gov/pubmed/36012291 http://dx.doi.org/10.3390/ijms23169025 |
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