Adenoviral overexpression of the glutamylcysteine ligase catalytic subunit protects pancreatic islets against oxidative stress

Phuong Oanh T. Tran, Sarah M. Parker, Eric LeRoy, Christopher C. Franklin, Terrance J. Kavanagh, Tao Zhang, Huarong Zhou, Portia Vliet, Elizabeth Oseid, Jamie S. Harmon, R. Paul Robertson

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44 Scopus citations

Abstract

The catalytic subunit of glutamylcysteine ligase (GCLC) primarily regulates de novo synthesis of glutathione (GSH) in mammalian cells and is central to the antioxidant capacity of the cell. However, GCLC expression in pancreatic islets has not been previously examined. We designed experiments to ascertain whether GCLC is normally expressed in islets and whether it is up-regulated by interleukin-1β (IL-1β). GCLC expression levels were intermediate compared with other metabolic tissues (kidney, liver, muscle, fat, and lung). IL-1β up-regulated GCLC expression (10 ng/ml IL-1β, 3.76 ± 0.86; 100 ng/ml IL-1β, 4.22 ± 0.68-fold control) via the p38 form of mitogen-activated protein kinase and NFκB and also increased reactive oxygen species levels (10 ng/ml IL-1β, 5.41 ± 1.8-fold control). This was accompanied by an increase in intraislet GSH/GSSG ratio (control, 7.1 ± 0.1; 10 ng/ml IL-1β, 8.0 ± 0.5; 100 ng/ml IL-1β, 8.2 ± 0.5-fold control; p < 0.05). To determine whether overexpression of GCLC increases the antioxidant capacity of the islet and prevents the adverse effects of IL-1β on glucose-induced insulin secretion, islets were infected with an adenovirus encoding GCLC. IL-1β significantly decreased glucose-stimulated insulin secretion (control, 123.8 ± 17.7; IL-1β, 40.2 ± 3.9 microunits/ml insulin/islet). GCLC overespression increased intraislet GSH levels and partially prevented the decrease in glucose-stimulated insulin secretion caused by IL-1β. These data provide the first report of GCLC expression in the islet and demonstrate that adenoviral overexpression of GCLC increases intracellular GSH levels and protects the beta cell from the adverse effects of IL-1β.

Original languageEnglish (US)
Pages (from-to)53988-53993
Number of pages6
JournalJournal of Biological Chemistry
Volume279
Issue number52
DOIs
StatePublished - Dec 24 2004

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    Tran, P. O. T., Parker, S. M., LeRoy, E., Franklin, C. C., Kavanagh, T. J., Zhang, T., Zhou, H., Vliet, P., Oseid, E., Harmon, J. S., & Robertson, R. P. (2004). Adenoviral overexpression of the glutamylcysteine ligase catalytic subunit protects pancreatic islets against oxidative stress. Journal of Biological Chemistry, 279(52), 53988-53993. https://doi.org/10.1074/jbc.M404809200