Differential roles of 3H-1,2-dithiole-3-thione-induced glutathione, glutathione S-transferase and aldose reductase in protecting against 4-hydroxy-2-nonenal toxicity in cultured cardiomyocytes
Section snippets
Materials
D3T with a purity of 99.8% was generously provided by Dr. Mary Tanga at SRI International (Menlo Park, CA) and Dr. Linda Brady at National Institute of Mental Health (Bethesda, MD). 4-Hydroxy-2-nonenal (HNE) was from Cayman Chemical (Ann Arbor, MI). Sorbinil was a generous gift from Pfeizer (Groton, CT). Dulbecco’s modified Eagle’s medium (DMEM), penicillin, streptomycin, and fetal bovine serum (FBS) were from Gibco-Invitrogen (Carlsbad, CA). All other chemicals and reagents were from Sigma
Elevation of cellular GSH and GST by D3T treatment
Incubation of H9C2 cardiomyocytes with D3T for 24 h resulted in significantly increased levels of both GSH and GST in a concentration- and time-dependent manner (Fig. 1, Fig. 2). GSH level was elevated by 25% at 1 h after treatment of the cells with 100 μM D3T; longer time incubation with D3T led to further increases in GSH level, which reached a maximum 2.5-fold induction at 24 h (Fig. 2A). A maximum elevation of the mRNA level for γGCL catalytic subunit was observed at 1 h; longer time incubation
Discussion
A number of cardiac disorders, including myocardial ischemia–reperfusion injury, congestive heart failure, and diabetic cardiomyopathy, have been linked to increases in ROS generation and lipid peroxidation within myocardium [1], [2], [3], [4]. It is therefore important to characterize enzymes that play a role in controlling the levels of prooxidant species and lipid peroxidation products, particularly the electrophilic aldehydes in cardiomyocytes. Although 1,2-dithiole-3-thiones, especially
Acknowledgments
This work was supported in part by NIH Grants CA91895 and HL71190 (Y.L.). M.A.T. was supported by NIH Grants ES03760, ES03819, and ES08078.
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