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Glutathione and antioxidant enzymes serve complementary roles in protecting activated hepatic stellate cells against hydrogen peroxide-induced cell death.
Biochim Biophys Acta. 2013 Dec; 1832(12):2027-34.BB

Abstract

BACKGROUND

In chronic liver disease, hepatic stellate cells (HSCs) are activated, highly proliferative and produce excessive amounts of extracellular matrix, leading to liver fibrosis. Elevated levels of toxic reactive oxygen species (ROS) produced during chronic liver injury have been implicated in this activation process. Therefore, activated hepatic stellate cells need to harbor highly effective anti-oxidants to protect against the toxic effects of ROS.

AIM

To investigate the protective mechanisms of activated HSCs against ROS-induced toxicity.

METHODS

Culture-activated rat HSCs were exposed to hydrogen peroxide. Necrosis and apoptosis were determined by Sytox Green or acridine orange staining, respectively. The hydrogen peroxide detoxifying enzymes catalase and glutathione-peroxidase (GPx) were inhibited using 3-amino-1,2,4-triazole and mercaptosuccinic acid, respectively. The anti-oxidant glutathione was depleted by L-buthionine-sulfoximine and repleted with the GSH-analogue GSH-monoethylester (GSH-MEE).

RESULTS

Upon activation, HSCs increase their cellular glutathione content and GPx expression, while MnSOD (both at mRNA and protein level) and catalase (at the protein level, but not at the mRNA level) decreased. Hydrogen peroxide did not induce cell death in activated HSCs. Glutathione depletion increased the sensitivity of HSCs to hydrogen peroxide, resulting in 35% and 75% necrotic cells at 0.2 and 1mmol/L hydrogen peroxide, respectively. The sensitizing effect was abolished by GSH-MEE. Inhibition of catalase or GPx significantly increased hydrogen peroxide-induced apoptosis, which was not reversed by GSH-MEE.

CONCLUSION

Activated HSCs have increased ROS-detoxifying capacity compared to quiescent HSCs. Glutathione levels increase during HSC activation and protect against ROS-induced necrosis, whereas hydrogen peroxide-detoxifying enzymes protect against apoptotic cell death.

Authors+Show Affiliations

Department of Gastroenterology and Hepatology, University Medical Center Groningen, University of Groningen, Groningen, The Netherlands.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

23871839

Citation

Dunning, Sandra, et al. "Glutathione and Antioxidant Enzymes Serve Complementary Roles in Protecting Activated Hepatic Stellate Cells Against Hydrogen Peroxide-induced Cell Death." Biochimica Et Biophysica Acta, vol. 1832, no. 12, 2013, pp. 2027-34.
Dunning S, Ur Rehman A, Tiebosch MH, et al. Glutathione and antioxidant enzymes serve complementary roles in protecting activated hepatic stellate cells against hydrogen peroxide-induced cell death. Biochim Biophys Acta. 2013;1832(12):2027-34.
Dunning, S., Ur Rehman, A., Tiebosch, M. H., Hannivoort, R. A., Haijer, F. W., Woudenberg, J., van den Heuvel, F. A., Buist-Homan, M., Faber, K. N., & Moshage, H. (2013). Glutathione and antioxidant enzymes serve complementary roles in protecting activated hepatic stellate cells against hydrogen peroxide-induced cell death. Biochimica Et Biophysica Acta, 1832(12), 2027-34. https://doi.org/10.1016/j.bbadis.2013.07.008
Dunning S, et al. Glutathione and Antioxidant Enzymes Serve Complementary Roles in Protecting Activated Hepatic Stellate Cells Against Hydrogen Peroxide-induced Cell Death. Biochim Biophys Acta. 2013;1832(12):2027-34. PubMed PMID: 23871839.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Glutathione and antioxidant enzymes serve complementary roles in protecting activated hepatic stellate cells against hydrogen peroxide-induced cell death. AU - Dunning,Sandra, AU - Ur Rehman,Atta, AU - Tiebosch,Marjolein H, AU - Hannivoort,Rebekka A, AU - Haijer,Floris W, AU - Woudenberg,Jannes, AU - van den Heuvel,Fiona A J, AU - Buist-Homan,Manon, AU - Faber,Klaas Nico, AU - Moshage,Han, Y1 - 2013/07/16/ PY - 2013/01/29/received PY - 2013/07/05/revised PY - 2013/07/09/accepted PY - 2013/7/23/entrez PY - 2013/7/23/pubmed PY - 2014/2/22/medline KW - 3-amino-1,2,4-triazole KW - 3AT KW - BSO KW - Catalase KW - Cell death KW - EDTA KW - FCS KW - GCL KW - GPx KW - GSH KW - GSH-MEE KW - GSH-monoethylester KW - GSSG KW - Glutathione peroxidase KW - HEPES KW - HO-1 KW - HSC KW - MS KW - N-2-hydroxyethylpiperazine-N-2-ethanesulfonic acid KW - NADPH-oxidase KW - Oxidative stress KW - PBS KW - PCR KW - PMSF KW - PPAR-γ KW - RNA KW - ROS KW - SOD KW - Superoxide dismutase KW - TGF-β KW - dNTP KW - deoxynucleoside triphosphates KW - ethylenediaminetetraacetic acid KW - fetal calf serum KW - glutamate cysteine ligase KW - glutathione peroxidase KW - heme-oxygenase-1 KW - hepatic stellate cell KW - l-buthionine-sulfoximine KW - mercaptosuccinic acid KW - nicotinamide adenine dinucleotide phosphate-oxidase KW - oxidized glutathione KW - peroxisome proliferator-activated receptor gamma KW - phenylmethanesulphonylfluoride KW - phosphate-buffered saline KW - polymerase chain reaction KW - reactive oxygen species KW - reduced glutathione KW - ribonucleic acid KW - super oxide dismutase KW - transforming growth factor beta KW - α-SMA KW - α-smooth muscle actin SP - 2027 EP - 34 JF - Biochimica et biophysica acta JO - Biochim Biophys Acta VL - 1832 IS - 12 N2 - BACKGROUND: In chronic liver disease, hepatic stellate cells (HSCs) are activated, highly proliferative and produce excessive amounts of extracellular matrix, leading to liver fibrosis. Elevated levels of toxic reactive oxygen species (ROS) produced during chronic liver injury have been implicated in this activation process. Therefore, activated hepatic stellate cells need to harbor highly effective anti-oxidants to protect against the toxic effects of ROS. AIM: To investigate the protective mechanisms of activated HSCs against ROS-induced toxicity. METHODS: Culture-activated rat HSCs were exposed to hydrogen peroxide. Necrosis and apoptosis were determined by Sytox Green or acridine orange staining, respectively. The hydrogen peroxide detoxifying enzymes catalase and glutathione-peroxidase (GPx) were inhibited using 3-amino-1,2,4-triazole and mercaptosuccinic acid, respectively. The anti-oxidant glutathione was depleted by L-buthionine-sulfoximine and repleted with the GSH-analogue GSH-monoethylester (GSH-MEE). RESULTS: Upon activation, HSCs increase their cellular glutathione content and GPx expression, while MnSOD (both at mRNA and protein level) and catalase (at the protein level, but not at the mRNA level) decreased. Hydrogen peroxide did not induce cell death in activated HSCs. Glutathione depletion increased the sensitivity of HSCs to hydrogen peroxide, resulting in 35% and 75% necrotic cells at 0.2 and 1mmol/L hydrogen peroxide, respectively. The sensitizing effect was abolished by GSH-MEE. Inhibition of catalase or GPx significantly increased hydrogen peroxide-induced apoptosis, which was not reversed by GSH-MEE. CONCLUSION: Activated HSCs have increased ROS-detoxifying capacity compared to quiescent HSCs. Glutathione levels increase during HSC activation and protect against ROS-induced necrosis, whereas hydrogen peroxide-detoxifying enzymes protect against apoptotic cell death. SN - 0006-3002 UR - https://www.unboundmedicine.com/medline/citation/23871839/Glutathione_and_antioxidant_enzymes_serve_complementary_roles_in_protecting_activated_hepatic_stellate_cells_against_hydrogen_peroxide_induced_cell_death_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0925-4439(13)00248-2 DB - PRIME DP - Unbound Medicine ER -