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Expression of bovine and mouse endothelial cell antioxidant enzymes following TNF-alpha exposure
J B Shaffer1, C P Treanor, P J Del Vecchio
1Wadsworth Center for Laboratories and Research, New York State Department of Health, Albany 12201.
Abstract:
Endothelial cells are primary targets for injury by reactive oxygen species. Endothelial catalase, copper-zinc superoxide dismutase (CuZnSOD), and manganous superoxide dismutase (MnSOD) provide potential antioxidant enzymatic defenses against oxidant-induced cellular damage. Previous studies in vivo and in vitro have demonstrated that in certain cell types exposure to oxidants may increase the expression of one or more of these antioxidant enzymes, thus providing greater intracellular potential to withstand oxidant-induced cell stress. To test whether endothelial antioxidant enzyme expression is influenced by similar oxidant-induced stresses in vitro, we have exposed endothelial cells to tumor necrosis factor-alpha (TNF-alpha) and have measured levels of catalase, CuZnSOD and MnSOD mRNA, and protein. Our results demonstrate a selective increase of MnSOD mRNA, with coordinate increases of both MnSOD protein and enzyme activity in endothelial cells treated for 24/h with TNF-alpha. In contrast, levels of catalase and CuZnSOD mRNA and protein remained unchanged in these cells after TNF-alpha treatment. These observations were made in microvessel endothelial cells derived from murine and bovine sources. Our results indicate that TNF-alpha can act specifically to increase enzymatic antioxidant potential in endothelial cells by induction of a particular antioxidant enzyme encoding mRNA species. These data demonstrate the capacity of endothelial cells to mount an antioxidant defense in response to exposure to an inducer of oxidative damage.
Insights
Tumor necrosis factor-alpha (TNF-alpha) selectively increases manganous superoxide dismutase (MnSOD) in endothelial cells, enhancing their antioxidant defense against reactive oxygen species. This response boosts cellular protection against oxidant-induced damage.
Area of Science:
- Cellular biology
- Biochemistry
- Oxidative stress research
Background:
- Endothelial cells are vulnerable to reactive oxygen species (ROS) damage.
- Antioxidant enzymes like catalase, CuZnSOD, and MnSOD offer cellular protection.
- Oxidant exposure can upregulate antioxidant enzyme expression in some cell types.
Purpose of the Study:
- To investigate if tumor necrosis factor-alpha (TNF-alpha) influences antioxidant enzyme expression in endothelial cells.
- To determine the specific effects of TNF-alpha on catalase, CuZnSOD, and MnSOD mRNA and protein levels.
Main Methods:
- Endothelial cells (murine and bovine microvessel) were exposed to TNF-alpha in vitro.
- Levels of catalase, CuZnSOD, and MnSOD mRNA and protein were measured.
- Enzyme activity assays were performed.
Main Results:
- TNF-alpha selectively increased MnSOD mRNA levels in endothelial cells.
- A coordinate increase in MnSOD protein and enzyme activity was observed.
- Catalase and CuZnSOD mRNA and protein levels remained unchanged.
Conclusions:
- TNF-alpha specifically enhances the antioxidant potential of endothelial cells.
- This enhancement is mediated by the induction of MnSOD mRNA.
- Endothelial cells can activate specific antioxidant defenses against inducers of oxidative damage.