Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2020 Nov;25(6):919-928.
doi: 10.1007/s12192-020-01127-8. Epub 2020 Jun 4.

Effect of di(2-ethylhexyl) phthalate on Nrf2-regulated glutathione homeostasis in mouse kidney

Ines Amara  1   2 , Amal Salah  1 , Rim Timoumi  1 , Emna Annabi  1 , Maria Scuto  2 , Angela Trovato  2 , Fadwa Neffati  3 , Vittorio Calabrese  2 , Salwa Abid-Essefi  4
Affiliations

Effect of di(2-ethylhexyl) phthalate on Nrf2-regulated glutathione homeostasis in mouse kidney

Ines Amara et al. Cell Stress Chaperones. 2020 Nov.

Abstract

Environmental toxicants such as phthalate have been involved in multiple health disorders including renal diseases. Oxidative damage is implicated in many alterations caused by phthalate especially the di(2-ethylhexyl) phthalate (DEHP), which is the most useful phthalate. However, information regarding its mechanism of renal damage is lacking. The transcription factor nuclear factor erythroid 2-related factor 2 (Nrf2) regulates gene expression implicated in free radical scavenging and cytoprotection including the antioxidant glutathione (GSH) pathway. The aim of this study was to assess whether DEHP affects the Nrf2 pathway and the GSH concentration. Mice were divided into four groups: a control group and three groups treated with DEHP at different concentrations (5, 50, and 200 mg/kg body weight) for 30 days. Our results showed that DEHP altered the normal levels of serum biochemical parameters creatinine (CREA), urea, and lactate dehydrogenase (LDH). This phthalate caused oxidative damage through the induction of lipid peroxidation and protein oxidation as marked by increase of protein carbonyl (PC) and loss of protein-bound sulfhydryls (PSH). Simultaneously, DEHP treatment decreased the protein level of Nrf-2, HO-1, and GCLC (responsible of GSH synthesis) and decreased the GSH level. Inhibition of the Nrf2 pathway is related to the activation of the mitochondrial pathway of apoptosis. This apoptotic process is evidenced by an upregulation of p53 and Bax protein levels in addition to a downregulation of Bcl-2. Collectively, our data demonstrated that depletion of Nrf2 and GSH was associated with the elevation of oxidative stress and the activation of intrinsic apoptosis in mouse kidney treated with DEHP.

Keywords: Apoptosis; Di(2-ethylhexyl) phthalate; Glutathione homeostasis; Nrf2 antioxidant pathway; Oxidative stress.

PubMed Disclaimer

Conflict of interest statement

The authors declare that there are no conflicts of interest.

Figures

Fig. 1
Fig. 1
Effect of DEHP treatment on malondihaldéhyde (MDA) (μmol MDA/mg of proteins) in kidney tissues of experimental mice after treatment of 30 days. Values represent mean ± SD (six animals were treated per group); *P < 0.05, **P < 0.01, and ***P < 0.001, values are significantly different from the control group
Fig. 2
Fig. 2
a Effect of DEHP treatment on carbonyl protein groups (nmol of carbonyl group/mg of protein) in kidney tissues of experimental mice after treatment of 30 days. Values represent mean ± SD (six animals were treated per group); **P < 0.01 and ***P < 0.001, values are significantly different from the control group. b Effect of DEHP treatment on protein-bound sulfhydryl (PSH) content (μg of PSH/mg of proteins) in kidney tissues of experimental mice after treatment of 30 days. Values represent mean ± SD (six animals were treated per group); **P < 0.01 and ***P < 0.001, values are significantly different from the control group
Fig. 3
Fig. 3
Inhibition of the Nrf2/HO-1/GCLC antioxidant pathway by DEHP in kidney tissues of experimental mice after treatment of 30 days. a Subcellular distribution of Nrf2 determined by Western blot analysis. β-Actin and Lamin B1 were served as loading controls for the cytosolic and nuclear fractions respectively. Data were collected from three independent experiments performed in replicate. b Protein levels of HO-1 and GCLC were analyzed by western blot. β-Actin was used as a loading control. RD: relative density as described in the “Material and methods” section. Values represent mean ± SD of three independent experiments. *P < 0.05 values are significantly different from the control group
Fig. 4
Fig. 4
Effect of DEHP treatment on GSH/GSSG ratio in kidney tissues of experimental mice after treatment of 30 days. Values represent mean ± SD (six animals were treated per group); *P < 0.05, **P < 0.01, and ***P < 0.001, values are significantly different from the control group
Fig. 5
Fig. 5
DEHP induces apoptosis in kidney of experimental mice after treatment of 30 days. Apoptosis markers P53, Bax, and Bcl2 were analyzed by western blot. β-Actin was used as a loading control. RD: relative density as described in the “Material and methods” section. Data are expressed as the mean ± SD of three separate experiments. *P < 0.05 values are significantly different from the control group
See this image and copyright information in PMC

References

    1. Al-Saleh I, Elkhatib R, Al-Rajoudi T, Al-Qudaihi G. Assessing the concentration of phthalate esters (PAEs) and bisphenol A (BPA) and the genotoxic potential of treated wastewater (final effluent) in Saudi Arabia. Sci Total Environ. 2017;578:440–451. - PubMed
    1. Anzovino A, Chiang S, Brown BE, Hawkins CL, Richardson DR, Huang ML. Molecular alterations in a mouse cardiac model of friedreich ataxia: an impaired Nrf2 response mediated via upregulation of Keap1 and activation of the Gsk3beta axis. Am J Pathol. 2017;187:2858–2875. - PubMed
    1. Atessahin A, Ceribasi AO, Yuce A, Bulmus O, Cikim G. Role of ellagic acid against cisplatin-induced nephrotoxicity and oxidative stress in rats. Basic Clin Pharmacol Toxicol. 2007;100:121–126. - PubMed
    1. Baird L, Dinkova-Kostova AT. The cytoprotective role of the Keap1-Nrf2 pathway. Arch Toxicol. 2011;85:241–272. - PubMed
    1. Beltifa A, Feriani A, Macherki M, Ghorbel A, Ghazouani L, Di Bella G, Sire O, Van Loco J, Reyns T, Mansour HB. Persistent plasticizers and bisphenol in the cheese of Tunisian markets induced biochemical and histopathological alterations in male BALB/c mice. Environ Sci Pollut Res Int. 2018;25:6545–6557. - PubMed

Publication types

MeSH terms