Secreted APE1/Ref-1 inhibits doxorubicin-induced cardiotoxicity via suppressing ROS generation and p53 pathway

Abstract Funding Acknowledgements Type of funding sources: Public grant(s) – National budget only. Main source(s): This research is supported by a grant the Korea Health Technology R&D Project through Industry Development Institute (KHIDI) funded Ministry & Welfare and Basic Science Research Program Foundation (NRF) Education, Republic Korea. Background Doxorubicin an anthracycline anticancer drug with potent therapeutic effect against various malignancies. However, its use limited due to cardiotoxicity, which can lead heart failure. Oxidative stress key player in development cardiotoxicity. Apurinic/apyrimidinic endonuclease 1/redox factor-1(APE1/Ref-1) multifunctional protein that regulates oxidative anti-inflammatory function. Secreted APE1/Ref-1 has shown anti-oxidant properties, but potential diseases largely unknown. Purpose study aims test protective secreted doxorubicin-induced cardiomyopathy model vivo vitro. Methods H9C2 cells were treated doxorubicin(2uM) induce Adenovirus preprotrypsin leading sequence APE1/Ref-1(AdPPT-LS-APE1/Ref-1), secretory adenovirus encoding human gene, was amplified HEK293T cells. then supernatant AdPPT-LS-APE1/Ref-1. In cardiotoxicity established injecting doxorubicin(15mg/kg) into C57BL/6 mice via peritoneum. The conditioned media AdPPT-LS-APE1/Ref-1 transfection injected i.p. after 24h. severity evaluated echocardiography plasma NT-proBNP. Cell survival, morphology, ROS level examined. expression cellular APE1/Ref-1, p53, Bax/Bcl-2, Caspase-3 analyzed Western blot ELISA. Redox activity measured using DTNB reduction assay. Results caused apoptosis mice. NT-proBNP increased both vitro(p<0.001) vivo(p<0.05) upon administering doxorubicin. Interestingly, doxorubicin p53 dose-dependent manner 48 hr, did not change. conditional showed redox decreased doxorubicin-treated mice(p<0.001 p<0.05, respectively). aggravated cell death(p<0.001) tissue damage(p<0.001) cells, while pre-treatment inhibited death cells(p<0.05). significantly reduced p53(p<0.001) effectors, Bax/Bcl-2(p<0.05) Caspase-3(p<0.05) It also p53(p<0.05) along mice(n=4 each group). Finally, intracellular production(p<0.001). Conclusion cardiomyocyte suppressing generation pathway. for prevention treatment