Dataset Information

EFFECTS OF ERYTHRODIOL ON THE ANTIOXIDANT RESPONSE AND PROTEOME OF HepG2 CELLS

ABSTRACT: Erythrodiol (EO) is a pentacyclic triterpenic alcohol found in the leaf of olive tree and in the olive oil. In this study, we evaluate the nature of cytotoxicity of EO on HepG2 hepatocarcinoma human cells studying the changes induced on cell viability, reactive oxygen species (ROS) production, antioxidant defence systems and proteome. The results obtained reveal that EO markedly decrease HepG2 cell viability without change ROS levels. A significant reduction in the glutathione and NADPH concentrations are produced with selective changes in the activity of antioxidant enzymes (glutathione peroxidase, glutathione reductase, glucose 6-phosphate dehydrogenase and 6-phosphogluconate dehydrogenase). Proteomics data reveals that 41 and 3 proteins are decreased completely or partially in abundance, respectively; and 29 are completely increased. The functional enrichment analysis showed that an impairment of important metabolic processes and in the nuclear transport of mature mRNA is produced while the biosynthesis of AMP and cell cycle G2/M phase transition are induced. The transcription factors and miRNA implied in this response are also identified

INSTRUMENT(S):

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Hepatocyte, Cell Culture

SUBMITTER: Carlos Alberto Fuentes Almagro

LAB HEAD: Juan Peragon Sanchez

PROVIDER: PXD029865 | Pride | 2022-02-17

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	JPeragon_sol26-17_C1_19072017.raw	Raw
	JPeragon_sol26-17_C2_19072017.raw	Raw
	JPeragon_sol26-17_C3_19072017.raw	Raw
	JPeragon_sol26-17_EO1_19072017.raw	Raw
	JPeragon_sol26-17_EO2_19072017.raw	Raw

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Publications

Effects of Erythrodiol on the Antioxidant Response and Proteome of HepG2 Cells.

Peñas-Fuentes Juan Luis JL Siles Eva E Rufino-Palomares Eva E EE Pérez-Jiménez Amalia A Reyes-Zurita Fernando J FJ Lupiáñez José A JA Fuentes-Almagro Carlos C Peragón-Sánchez Juan J

Antioxidants (Basel, Switzerland) 20211229 1

Erythrodiol (EO) is a pentacyclic triterpenic alcohol found in olive tree leaves and olive oil, and it has important effects on the health properties and quality of olive oil. In this study, we characterized the cytotoxic effects of EO on human hepatocarcinoma (HepG2) cells by studying changes in cell viability, reactive oxygen species (ROS) production, antioxidant defense systems, and the proteome. The results reveal that EO markedly decreased HepG2 cell viability without changing ROS levels. T ...[more]

PMID: 35052578

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Project description:With an increasing focus in science, politics and society on animal welfare, human in vitro systems are gaining particular interest as potential replacement tools for animal experimentation. However, animal components are still commonly used in cell culture models, thereby compromising the true potential of in vitro systems as “pure” animal replacement tools. Especially fetal bovine serum (FBS) is heavily used in cell culture models, sparking debates about the ethical and scientific defensibleness of the usage of FBS in in vitro cell culture systems. In this study, we performed a comparative proteomic characterization of HepG2 cells adapted to serum-free conditions with cells cultivated under FBS-containing conditions. Four different serum-free media were used, of which two were previously shown to support long-term serum-free cultivation of HepG2 cells. Notably, while two media can be characterized as xeno-free, two media still contain animal components (bovine transferrin and/or bovine serum albumin). As we have previously shown increase sensitivity of serum-free cultivated HepG2 cells in cytotoxicity assays, as well as increased oxidative stress protection, we set a particular focus on drug-metabolizing enzymes and NRF2 regulated proteins. Using principal component analysis and pathways analysis, we identified nutrient related stress and energy generation as key factors for survival under serum-free conditions, either due to a decreased/altered availability of substrates for utilization or missing micronutrients. Furthermore, we observed predicted up-regulation of multiple pathways associated with drug metabolism and oxidative stress protection, including; “phase I – functionalization of compounds”, “phase II – conjugation of compounds” and “NRF2 mediated oxidative stress response”. We observed significant up-regulation of multiple members of the glutathione-s-transferase and glutathione peroxidase families and confirmed that increased antioxidant enzyme expression correlates with higher enzyme activity in vitro. These findings support our previously generated results derived from different toxicity assays and provide insights into the changes inflicted upon HepG2 cells by altered culturing conditions.