Open AccessIdentification of protein targets underlying dietary nitrate‐induced protection against doxorubicin cardiotoxicity
Author(s) -
Xi Lei,
Zhu ShuGuang,
Hobbs Daniel C.,
Kukreja Rakesh C.
Publication year - 2011
Publication title -
journal of cellular and molecular medicine
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.44
H-Index - 130
eISSN - 1582-4934
pISSN - 1582-1838
DOI - 10.1111/j.1582-4934.2011.01257.x
Subject(s) - nitrate , chemistry , doxorubicin , pharmacology , cardiotoxicity , cardioprotection , superoxide dismutase , antioxidant , nitrate reductase , peroxiredoxin , western blot , biochemistry , enzyme , biology , toxicity , medicine , chemotherapy , ischemia , peroxidase , organic chemistry , gene
We recently demonstrated protective effect of chronic oral nitrate supplementation against cardiomyopathy caused by doxorubicin (DOX), a highly effective anticancer drug. The present study was designed to identify novel protein targets related to nitrate‐induced cardioprotection. Adult male CF‐1 mice received cardioprotective regimen of nitrate (1 g NaNO 3 per litre of drinking water) for 7 days before DOX injection (15 mg/kg, i.p.) and continued for 5 days after DOX treatment. Subsequently the heart samples were collected for proteomic analysis with two‐dimensional differential in‐gel electrophoresis with 3 CyDye labelling. Using 1.5 cut‐off ratio, we identified 36 proteins that were up‐regulated by DOX in which 32 were completely reversed by nitrate supplementation (89%). Among 19 proteins down‐regulated by DOX, 9 were fully normalized by nitrate (47%). The protein spots were further identified with Matrix Assisted Laser Desorption/Ionization‐Time‐of‐Flight (MALDI‐TOF)/TOF tandem mass spectrometry. Three mitochondrial antioxidant enzymes were altered by DOX, i.e. up‐regulation of manganese superoxide dismutase and peroxiredoxin 3 (Prx3), and down‐regulation of Prx5, which were reversed by nitrate. These results were further confirmed by Western blots. Nitrate supplementation also significantly improved animal survival rate from 80% in DOX alone group to 93% in Nitrate + DOX group 5 days after the DOX treatment. In conclusion, the proteomic analysis has identified novel protein targets underlying nitrate‐induced cardioprotection. Up‐regulation of Prx5 by nitrate may explain the observed enhancement of cardiac antioxidant defence by nitrate supplementation.