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. 2013 Dec;13(4):347-56.
doi: 10.1007/s12012-013-9215-1.

Protection from oxidative and electrophilic stress in the Gsta4-null mouse heart

Helen Beneš  1 , Mai K Vuong, Marjan Boerma, Kevin E McElhanon, Eric R Siegel, Sharda P Singh
Affiliations

Protection from oxidative and electrophilic stress in the Gsta4-null mouse heart

Helen Beneš et al. Cardiovasc Toxicol. 2013 Dec.

Abstract

4-Hydroxynonenal (4-HNE) mediates many pathological effects of oxidative and electrophilic stress and signals to activate cytoprotective gene expression regulated by NF-E2-related factor 2 (Nrf2). By exhibiting very high levels of 4-HNE-conjugating activity, the murine glutathione transferase alpha 4 (GSTA4-4) helps regulate cellular 4-HNE levels. To examine the role of 4-HNE in vivo, we disrupted the murine Gsta4 gene. Gsta4-null mice exhibited no cardiac phenotype under normal conditions and no difference in cardiac 4-HNE level as compared to wild-type mice. We hypothesized that the Nrf2 pathway might contribute an important compensatory mechanism to remove excess cardiac 4-HNE in Gsta4-null mice. Cardiac nuclear extracts from Gsta4-null mice exhibited significantly higher Nrf2 binding to antioxidant response elements. We also observed responses in critical Nrf2 target gene products: elevated Sod2, Cat, and Akr1b7 mRNA levels and significant increases in both cardiac antioxidant and anti-electrophile enzyme activities. Gsta4-null mice were less sensitive and maintained normal cardiac function following chronic doxorubicin treatment, known to increase cardiac 4-HNE levels. Hence, in the absence of GSTA4-4 to modulate both physiological and pathological 4-HNE levels, the adaptive Nrf2 pathway may be primed to contribute to a preconditioned cardiac phenotype in the Gsta4-null mouse.

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Figures

Fig. 1
Fig. 1
No difference in cardiac 4-HNE levels between wild-type and Gsta4-null mice. 4-HNE levels were determined, as previously reported (23), in liver and cardiac tissue samples from 16-week-old male mice. The difference between the null and wild-type mice was only significant for the liver (*, p = 2×10 −4, n=4).
Fig. 2
Fig. 2
Different levels of active nuclear Nrf2 in hearts of wild-type and Gsta4-null mice. Active Nrf2 binding was measured in nuclear extracts from whole hearts of male WT and Gsta4-null mice (16 weeks of age). Open bars: nuclear extract; hatched bars: nuclear extract + mutated probe; cross-hatched bars: nuclear extract + competitive ARE probe. Each bar represents the mean ± SD (n=4); statistical significance, as shown, was evaluated by the t-test.
Fig. 3
Fig. 3
Anti-oxidant and anti-electrophile activities in WT and Gsta4 null (KO) mice. All enzyme activities were measured in heart extracts of 16-week old male mice as described in the Methods. Each bar represents the mean ± SD (n=4); statistical significance, as shown, was evaluated by a t-test. (A) Catalase activity was assayed using H2O2 as a substrate. (B) SOD enzyme activities were determined using WST1 (see Methods) as a substrate. (C) AKR and ALDH activities were measured using 4-HNE as a substrate.
Fig. 3
Fig. 3
Anti-oxidant and anti-electrophile activities in WT and Gsta4 null (KO) mice. All enzyme activities were measured in heart extracts of 16-week old male mice as described in the Methods. Each bar represents the mean ± SD (n=4); statistical significance, as shown, was evaluated by a t-test. (A) Catalase activity was assayed using H2O2 as a substrate. (B) SOD enzyme activities were determined using WST1 (see Methods) as a substrate. (C) AKR and ALDH activities were measured using 4-HNE as a substrate.
Fig. 3
Fig. 3
Anti-oxidant and anti-electrophile activities in WT and Gsta4 null (KO) mice. All enzyme activities were measured in heart extracts of 16-week old male mice as described in the Methods. Each bar represents the mean ± SD (n=4); statistical significance, as shown, was evaluated by a t-test. (A) Catalase activity was assayed using H2O2 as a substrate. (B) SOD enzyme activities were determined using WST1 (see Methods) as a substrate. (C) AKR and ALDH activities were measured using 4-HNE as a substrate.
Fig. 4
Fig. 4
Survival of Gsta4-null and WT mice treated with DOX. 8 WT and 8 Gsta4-null mice (male, 12–16 weeks of age) were injected weekly (indicated by arrows) with 5 mg/kg DOX for 4 weeks. Survival, analyzed using the log-rank test and Cox regression, indicated that Gsta4-null mice had a significant reduction in mortality (hazard ratio = 0.132; log-rank p = 0.033).
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References

    1. Heistad DD, Wakisaka Y, Miller J, Chu Y, Pena-Silva R. Novel aspects of oxidative stress in cardiovascular diseases. Circulation journal: official journal of the Japanese Circulation Society. 2009;73:201–207. - PMC - PubMed
    1. Ari E, Kaya Y, Demir H, Cebi A, Alp HH, Bakan E, Odabasi D, Keskin S. Oxidative DNA damage correlates with carotid artery atherosclerosis in hemodialysis patients. Hemodialysis international. 2011;15:453–459. - PubMed
    1. Fadel PJ, Farias M, III, Gallagher KM, Wang Z, Thomas GD. Oxidative stress and enhanced sympathetic vasoconstriction in contracting muscles of nitrate-tolerant rats and humans. J Physiol (Lond) 2012;590:395–407. - PMC - PubMed
    1. Hollander JM, Baseler WA, Dabkowski ER. Proteomic remodeling of mitochondria in heart failure. Congestive Heart Failure. 2011;17:262–268. - PMC - PubMed
    1. Schlage WK, Westra JW, Gebel S, Catlett NL, Mathis C, Frushour BP, Hengstermann A, Van Hooser A, Poussin C, Wong B, Lietz M, Park J, Drubin D, Veljkovic E, Peitsch MC, Hoeng J, Deehan R. A computable cellular stress network model for non-diseased pulmonary and cardiovascular tissue. BMC Syst Biol. 2011;5:168. - PMC - PubMed

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