Autophagy and protein kinase C are required for cardioprotection by sulfaphenazole.
Previously, we showed that sulfaphenazole (SUL), an antimicrobial agent that is a potent inhibitor of cytochrome P4502C9, is protective against ischemia-reperfusion (I/R) injury (Ref. 15). The mechanism, however, underlying this cardioprotection, is largely unknown. With evidence that activation of autophagy is protective against simulated I/R in HL-1 cells, and evidence that autophagy is upregulated in preconditioned hearts, we hypothesized that SUL-mediated cardioprotection might resemble ischemic preconditioning with respect to activation of protein kinase C and autophagy. We used the Langendorff model of global ischemia to assess the role of autophagy and protein kinase C in myocardial protection by SUL during I/R. We show that SUL enhanced recovery of function, reduced creatine kinase release, decreased infarct size, and induced autophagy. SUL also triggered PKC translocation, whereas inhibition of PKC with chelerythrine blocked the activation of autophagy in adult rat cardiomyocytes. In the Langendorff model, chelerythrine suppressed autophagy and abolished the protection mediated by SUL. SUL increased autophagy in adult rat cardiomyocytes infected with GFP-LC3 adenovirus, in isolated perfused rat hearts, and in mCherry-LC3 transgenic mice. To establish the role of autophagy in cardioprotection, we used the cell-permeable dominant-negative inhibitor of autophagy, Tat-Atg5(K130R). Autophagy and cardioprotection were abolished in rat hearts perfused with recombinant Tat-Atg5(K130R). Taken together, these studies indicate that cardioprotection mediated by SUL involves a PKC-dependent induction of autophagy. The findings suggest that autophagy may be a fundamental process that enhances the heart's tolerance to ischemia.
Huang
Chengqun
C
BioScience Center, San Diego State University, San Diego, California, USA.
Liu
Wayne
W
Perry
Cynthia N
CN
Yitzhaki
Smadar
S
Lee
Youngil
Y
Yuan
Hua
H
Tsukada
Yayoi Tetsuo
YT
Hamacher-Brady
Anne
A
Mentzer
Robert M
RM
Gottlieb
Roberta A
RA
D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D000890:Anti-Infective Agents / Q000494:pharmacology / Q000627:therapeutic use*; D001343:Autophagy / Q000187:drug effects / Q000502:physiology*; D000071187:Autophagy-Related Protein 5; D053119:Benzophenanthridines / Q000494:pharmacology; D002478:Cells, Cultured; D004195:Disease Models, Animal; D051379:Mice; D008822:Mice, Transgenic; D008869:Microtubule-Associated Proteins / Q000235:genetics / Q000378:metabolism; D015428:Myocardial Reperfusion Injury / Q000378:metabolism / Q000503:physiopathology / Q000517:prevention & control*; D032383:Myocytes, Cardiac / Q000378:metabolism / Q000473:pathology; D011493:Protein Kinase C / Q000378:metabolism*; D051745:Protein Kinase C-delta / Q000378:metabolism; D011506:Proteins / Q000494:pharmacology; D051381:Rats; D013426:Sulfaphenazole / Q000494:pharmacology / Q000627:therapeutic use*
D000900:Anti-Bacterial Agents; D000890:Anti-Infective Agents; D000071187:Autophagy-Related Protein 5; D053119:Benzophenanthridines; D008869:Microtubule-Associated Proteins; D011493:Protein Kinase C; D051745:Protein Kinase C-delta; D011506:Proteins; D013426:Sulfaphenazole; C545865:Atg5 protein, rat; C526109:Anxa3 protein, rat; C480860:Map1lc3b protein, mouse; C016299:chelerythrine