Abstract
Backgrounds: Nitric oxide (NO) plays a key role in cardioprotection. Its role against cardioprotection is dependent on the level of NO. Although it is well known that NO preconditioning has cardioprotective effects, but its mechanism remains unsatisfactory.
Methods: To induce NO preconditioning, H9c2 cells were treated with low NO concentration and subsequently induced apoptosis by high NO. The signalling and anti-apoptotic effects of NO preconditioning were monitored by Western blotting, facs analysis.
Results: Sodium nitroprusside (SNP)-induced cytotoxicity was inhibited by low SNP (0.3 mM) preconditioning. Furthermore, low SNP phosphorylated Akt/FoxO1 in the presence of high SNP (1.5 mM), while phosphorylated Akt/FoxO1 and viability were reversed by PI3K inhibitor. Also, low NO-induced CREB phosphorylation with high NO was inhibited by PKA inhibitor, indicating that NO preconditioning protects NO-induced cytotoxicity in PKA dependently. Apoptotic inhibition with NO preconditioning was accompanied with increased Bcl-2, decreased Bax, and caspase3 activation, which was all reversed by LY294002.
Conclusion: Our results indicate that low NO preconditioning prevent subsequent high NO-induced apoptosis via Akt/PKA/CREB activation in H9c2 cells.
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This work was supported by the National Research Foundation of Korea (NRF) grant funded by the Korea government (NRF-2017R1A2B1011688 and 2018R1A2A3075684).
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Hyun-Jeong Kwak, Jae-Young Um & Sang-Seob Lee declare that they have no competing interests.
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Kwak, HJ., Um, JY. & Lee, SS. Mild NO preconditioning protects H9c2 cells against NO-induced apoptosis through activation of PI3K/Akt and PKA-dependent pathways. Mol. Cell. Toxicol. 15, 297–305 (2019). https://doi.org/10.1007/s13273-019-0033-9
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DOI: https://doi.org/10.1007/s13273-019-0033-9