Activation of ATM/Akt/CREB/eNOS signaling axis by aphidicolin increases no production and vessel relaxation in endothelial cells and rat aortas

Jung Hyun Park, Du Hyong Cho, Yun Jin Hwang, Jee Young Lee, Hyeon Ju Lee, Inho Jo

Research output: Contribution to journalArticlepeer-review

2 Scopus citations

Abstract

Although DNA damage responses (DDRs) are reported to be involved in nitric oxide (NO) production in response to genotoxic stresses, the precise mechanism of DDR-mediated NO production has not been fully understood. Using a genotoxic agent aphidi-colin, we investigated how DDRs regulate NO production in bovine aortic endothelial cells. Prolonged (over 24 h) treatment with aphidicolin increased NO production and endothelial NO synthase (eNOS) protein expression, which was accompanied by increased eNOS dimer/monomer ratio, tetrahydrobiopterin levels, and eNOS mRNA expression. A promoter assay using 5’-serially deleted eNOS promoters revealed that Tax-responsive element site, located at −962 to −873 of the eNOS promoter, was respon-sible for aphidicolin-stimulated eNOS gene expression. Aphidicolin increased CREB activity and ectopic expression of dominant-negative inhibitor of CREB, A-CREB, repressed the stimulatory effects of aphidicolin on eNOS gene expression and its promoter activity. Co-treatment with LY294002 decreased the aphidicolin-stimulated increase in p-CREB-Ser133 level, eNOS expression, and NO production. Furthermore, ectopic expression of dominant-negative Akt construct attenuated aphidicolin-stimulated NO production. Aphidicolin increased p-ATM-Ser1981 and the knockdown of ATM using siRNA attenuated all stimulatory effects of aphidicolin on p-Akt-Ser473, p-CREB-Ser133, eNOS expression, and NO production. Additionally, these stimulatory effects of aphidi-colin were similarly observed in human umbilical vein endothelial cells. Lastly, aphidicolin increased acetylcholine-induced vessel relaxation in rat aortas, which was accompanied by increased p-ATM-Ser1981, p-Akt-Ser473, p-CREB-Ser133, and eNOS expression. In conclusion, our results demonstrate that in response to aphidicolin, activation of ATM/Akt/CREB/eNOS signaling cascade mediates increase of NO production and vessel relaxation in endothelial cells and rat aortas.

Original languageEnglish
Pages (from-to)549-560
Number of pages12
JournalBiomolecules and Therapeutics
Volume28
Issue number6
DOIs
StatePublished - 2020

Bibliographical note

Funding Information:
This work was supported by National Research Foundation grants (2018R1A2B2002062, 2018R1D1A1B07050732, and 2017R1D1A1B03034131) from the Korean government and an RP-Grant in 2016 from Ewha Womans University.

Publisher Copyright:
© 2020 The Korean Society of Applied Pharmacology.

Keywords

  • Aphidicolin
  • DNA damage response
  • Endothelial nitric oxide synthase
  • Nitric oxide
  • Vessel relaxation

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