Malondialdehyde and 4-hydroxynonenal adducts are not formed on cardiac ryanodine receptor (RyR2) and sarco(endo)plasmic
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Malondialdehyde and 4-hydroxynonenal adducts are not formed on cardiac ryanodine receptor (RyR2) and sarco(endo)plasmic reticulum Ca2+-ATPase (SERCA2) in diabetes Caronda J. Moore • Chun Hong Shao • Ryoji Nagai • Shelby Kutty • Jaipaul Singh Keshore R. Bidasee
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Received: 23 August 2012 / Accepted: 9 January 2013 / Published online: 25 January 2013 Ó Springer Science+Business Media New York 2013
Abstract Recently, we reported an elevated level of glucose-generated carbonyl adducts on cardiac ryanodine receptor (RyR2) and sarco(endo)plasmic reticulum Ca2?ATPase (SERCA2) in hearts of streptozotocin(STZ)induced diabetic rats. We also showed these adduct impaired RyR2 and SERCA2 activities, and altered evoked Ca2? transients. What is less clear is if lipid-derived malondialdehyde (MDA) and 4-hydroxy-2-nonenal (4-HNE) also chemically react with and impair RyR2 and SERCA2 activities in diabetes? This study used western blot assays C. J. Moore C. H. Shao K. R. Bidasee (&) Department of Pharmacology and Experimental Neuroscience, 985800 Nebraska Medical Center, Durham Research Center, DRC 3047, Omaha, NE 68198-5800, USA e-mail: [email protected] R. Nagai Laboratory of Food and Regulation Biology, Department of Biosciences, School of Agriculture, Tokai University, Tokyo, Japan S. Kutty Joint Division of Pediatric Cardiology, University of Nebraska/ Creighton University and Children’s Hospital and Medical Center, Omaha, NE, USA J. Singh School of Forensic and Investigative Sciences and School of Pharmacy and Biomedical Sciences, University of Central Lancashire, Preston, UK K. R. Bidasee Environmental, Agricultural and Occupational Health, University of Nebraska Medical Center, Omaha, NE 68198, USA K. R. Bidasee N146 Beadle Center, Nebraska Center for Redox Biology, Lincoln, NE 68588-0662, USA
with adduct-specific antibodies and confocal microscopy to assess levels of MDA, 4-HNE, Ne-carboxy(methyl)lysine (CML), pentosidine, and pyrraline adducts on RyR2 and SERCA2 and evoked intracellular transient Ca2? kinetics in myocytes from control, diabetic, and treated-diabetic rats. MDA and 4-HNE adducts were not detected on RyR2 and SERCA2 from either control or 8 weeks diabetic rats with altered evoked Ca2? transients. However, CML, pentosidine, and pyrraline adducts were elevated three- to five-fold (p \ 0.05). Treating diabetic rats with pyridoxamine (a scavenger of reactive carbonyl species, RCS) or aminoguanidine (a mixed reactive oxygen species-RCS scavenger) reduced CML, pentosidine, and pyrraline adducts on RyR2 and SERCA2 and blunted SR Ca2? cycling changes. Treating diabetic rats with the superoxide dismutase mimetic tempol had no impact on MDA and 4-HNE adducts on RyR2 and SERCA2, and on SR Ca2? cycling. From these data we conclude that lipid-derived MDA and 4-HNE adducts are not formed on RyR2 and SERCA2 in this model of diabetes, and are therefore unlikely to be directly contributing to the SR Ca2? dysregulation. Keywords Diabetes mellitus Rats Malondialdehyde 4-Hydroxynonenal Post-translational modificati
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