ALDH-2 deficiency increases cardiovascular oxidative stress---Evidence for indirect antioxidative properties

doi:10.1016/j.bbrc.2007.12.089

Biochemical and Biophysical Research Communications

Volume 367, Issue 1, 29 February 2008, Pages 137-143

https://doi.org/10.1016/j.bbrc.2007.12.089 Get rights and content

Abstract

Mitochondrial aldehyde dehydrogenase (ALDH-2) reduces reactive oxygen species (ROS) formation related to toxic aldehydes; additionally, it provides a bioactivating pathway for nitroglycerin. Since acetaldehyde, nitroglycerin, and doxorubicin treatment provoke mitochondrial oxidative stress, we used ALDH-2^−/− mice and purified recombinant human ALDH-2 to test the hypothesis that ALDH-2 has an indirect antioxidant function in mitochondria. Antioxidant capacity of purified ALDH-2 was comparable to equimolar doses of glutathione, cysteine, and dithiothreitol; mitochondrial oxidative stress was comparable in C57Bl6 and ALDH-2^−/− mice after acute challenges with nitroglycerin or doxorubicin, whereas chronic acetaldehyde, nitroglycerin, and doxorubicin treatment dose-dependently increased mitochondrial ROS formation and impaired endothelial function to a greater extent in ALDH-2^−/− mice. Maximal nitroglycerin dose applied in vivo lead to a “super-desensitized” nitroglycerin response in isolated ALDH-2^−/− aortas, inaccessible in C57Bl6 mice. Our results suggest that ALDH-2 has an indirect antioxidative property independent of its thiol-moiety in disease states of cardiovascular oxidative stress.

Section snippets

Materials and methods

Animals and in vivo treatment. Young adult (10weeks) male ALDH-2^−/− mice were age matched with C57Bl6 mice (wild type, WT). The generation of the ALDH-2 null mutant was described elsewhere [19]. Mice were equipped with micro osmotic pumps from Alzet (Cupertino, CA) containing 450 mM GTN solved in ethanol (undiluted, e.g. 100 μg/h/4d, diluted 1:1 with ethanol, e.g. 50 μg/h/4d, diluted 1:9, e.g. 10 μg/h/4d) or ethanol as a control. Mice were also treated in vivo with acetaldehyde (1.2 μl/d s.c. for 3

Activity and antioxidant capacities of human ALDH-2

The isolated and purified human ALDH-2 enzyme was tested with regard to its potential antioxidant properties. Compared to equimolar amounts of glutathione (GSH), dithiothreitol (DTT), and cysteine, ALDH-2 did not differ in its peroxynitrite scavenging properties (see Fig. 1A). Additionally, the ROS formation of isolated rat cardiac mitochondria from untreated Wistar rats was not increased upon in vitro inhibition of ALDH-2 by the selective inhibitor, daidzin (200 μM, see Fig. 1B). Finally, acute

Discussion

The main finding of our study is, that the mitochondrially located enzyme ALDH-2 seems to provide indirect antioxidant properties presumably by preventing the accumulation of toxic aldehydes rather than direct ROS scavenging activity based on its chemical structure. This explains best the observed more pronounced tolerance in response to in vivo nitroglycerin administration in ALDH-2^−/− mice compared to WT but lack of any significant differences upon acute challenges with acetaldehyde, GTN or

Acknowledgments

We thank Dr. Ho and Prof. Weiner (Purdue University, West Lafayette, USA) for providing the human recombinant ALDH-2 plasmid. The expert technical assistance of Jörg Schreiner is gratefully appreciated. The financial support by the German Research Foundation (SFB 553 to A.D. and T.M.), by MAIFOR grants from the University Hospital Mainz (A.D. and P.W.) and by the Robert-Müller-Foundation (T.M. and A.D.) is gratefully acknowledged. This report contains results that are part of the thesis work of

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Citation Excerpt :
Another mechanism by which DOX may cause elevated ROS levels is through dysregulation of antioxidant enzymes, such as glutathione peroxidase-1, which is readily inactivated upon exposure to DOX in cardiac tissue [9], while glutathione peroxidase-1 deficiency in mice was shown to increase the susceptibility to DOX-induced cardiotoxicity and overexpression of glutathione peroxidase-1 rescued the cardiac phenotype. In addition, it has been demonstrated that up-regulation of the mitochondrial antioxidant enzyme manganese superoxide dismutase (SOD2) counteracted cardiotoxic effects induced by DOX exposure [10], whereas deficiency in mitochondrial aldehyde dehydrogenase 2, which removes toxic aldehydes such as 4-hydroxynonenal resulting from lipid peroxidation, rendered mice more susceptible to DOX-induced cardiac and vascular damage [11]. Of note, genetic deletion of SOD2 in mice leads to prenatal or premature death of the animals, due to cardiomyocyte damage and dilated cardiomyopathy [12,13].
The anthracycline doxorubicin (DOX) is widely used in cancer therapy with the limitation of cardiotoxicity leading to the development of congestive heart failure. DOX-induced oxidative stress and changes of the phosphoproteome as well as epigenome were described but the exact mechanisms of the adverse long-term effects are still elusive. Here, we tested the impact of DOX treatment on cell death, oxidative stress parameters and expression profiles of proteins involved in epigenetic pathways in a cardiomyocyte cell culture model. Markers of oxidative stress, apoptosis and expression of proteins involved in epigenetic processes were assessed by immunoblotting in cultured rat myoblasts (H9c2) upon treatment with DOX (1 or 5 μM for 24 or 48 h) in adherent viable and detached apoptotic cells. The apoptosis markers cleaved caspase-3 and fractin as well as oxidative stress markers 3-nitrotyrosine and malondialdehyde were dose-dependently increased by DOX treatment. Histone deacetylases (SIRT1 and HDAC2), histone lysine demethylases (KDM3A and LSD1) and histone lysine methyltransferases (SET7 and SMYD1) were significantly regulated by DOX treatment with generation of cleaved protein fragments and posttranslational modifications. Overall, we found significant decrease in histone 3 acetylation in DOX-treated cells. DOX treatment of cultured cardiomyocyte precursor cells causes severe cell death by apoptosis associated with cellular oxidative stress. In addition, significant regulation of proteins involved in epigenetic processes and changes in global histone 3 acetylation were observed. However, the significance and clinical impact of these changes remain elusive.

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¹: These authors contributed equally to this work.

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ALDH-2 deficiency increases cardiovascular oxidative stress---Evidence for indirect antioxidative properties

Abstract

Section snippets

Materials and methods

Activity and antioxidant capacities of human ALDH-2

Discussion

Acknowledgments

Free Radic. Biol. Med.

Free Radic. Biol. Med.

Alcohol

J. Biol. Chem.

Free Radic. Biol. Med.

FEBS Lett.

Free Radic. Biol. Med.

Toxicol. Appl. Pharmacol.

Free Radic. Biol. Med.

Free Radic. Biol. Med.

Free Radic. Biol. Med.

J. Biol. Chem.

Free Radic. Biol. Med.

Identification of the enzymatic mechanism of nitroglycerin bioactivation

Proc. Natl. Acad. Sci. USA

Central role of mitochondrial aldehyde dehydrogenase and reactive oxygen species in nitroglycerin tolerance and cross-tolerance

J. Clin. Invest.

Mitochondrial aldehyde dehydrogenase-2 (ALDH2) Glu504Lys polymorphism contributes to the variation in efficacy of sublingual nitroglycerin

J. Clin. Invest.