Omeprazole induces heme oxygenase-1 in fetal human pulmonary microvascular endothelial cells via hydrogen peroxide-independent Nrf2 signaling pathway

Toxicol Appl Pharmacol. 2016 Nov 15;311:26-33. doi: 10.1016/j.taap.2016.10.002. Epub 2016 Oct 8.

Abstract

Omeprazole (OM) is an aryl hydrocarbon receptor (AhR) agonist and a proton pump inhibitor that is used to treat humans with gastric acid related disorders. Recently, we showed that OM induces NAD (P) H quinone oxidoreductase-1 (NQO1) via nuclear factor erythroid 2-related factor 2 (Nrf2)-dependent mechanism. Heme oxygenase-1 (HO-1) is another cytoprotective and antioxidant enzyme that is regulated by Nrf2. Whether OM induces HO-1 in fetal human pulmonary microvascular endothelial cells (HPMEC) is unknown. Therefore, we tested the hypothesis that OM will induce HO-1 expression via Nrf2 in HPMEC. OM induced HO-1 mRNA and protein expression in a dose-dependent manner. siRNA-mediated knockdown of AhR failed to abrogate, whereas knockdown of Nrf2 abrogated HO-1 induction by OM. To identify the underlying molecular mechanisms, we determined the effects of OM on cellular hydrogen peroxide (H2O2) levels since oxidative stress mediated by the latter is known to activate Nrf2. Interestingly, the concentration at which OM induced HO-1 also increased H2O2 levels. Furthermore, H2O2 independently augmented HO-1 expression. Although N-acetyl cysteine (NAC) significantly decreased H2O2 levels in OM-treated cells, we observed that OM further increased HO-1 mRNA and protein expression in NAC-pretreated compared to vehicle-pretreated cells, suggesting that OM induces HO-1 via H2O2-independent mechanisms. In conclusion, we provide evidence that OM transcriptionally induces HO-1 via AhR - and H2O2 - independent, but Nrf2 - dependent mechanisms. These results have important implications for human disorders where Nrf2 and HO-1 play a beneficial role.

Keywords: Aryl hydrocarbon receptor; Fetal human pulmonary microvascular endothelial cells; Heme oxygenase-1; Hydrogen peroxide; Nuclear factor erythroid 2-related factor 2; Omeprazole.

MeSH terms

  • Cells, Cultured
  • Endothelium, Vascular / drug effects
  • Endothelium, Vascular / enzymology
  • Heme Oxygenase-1 / biosynthesis*
  • Humans
  • Hydrogen Peroxide / metabolism
  • Hydrogen Peroxide / pharmacology*
  • Lung / blood supply
  • Lung / embryology*
  • Microvessels / drug effects*
  • Microvessels / enzymology
  • NF-E2-Related Factor 2 / metabolism*
  • Omeprazole / pharmacology*
  • Signal Transduction*

Substances

  • NF-E2-Related Factor 2
  • NFE2L2 protein, human
  • Hydrogen Peroxide
  • Heme Oxygenase-1
  • Omeprazole