Elsevier

Free Radical Biology and Medicine

Volume 51, Issue 10, 15 November 2011, Pages 1910-1917
Free Radical Biology and Medicine

Original Contribution
Omeprazole attenuates hyperoxic injury in H441 cells via the aryl hydrocarbon receptor

https://doi.org/10.1016/j.freeradbiomed.2011.08.013Get rights and content

Abstract

Hyperoxia contributes to the development of bronchopulmonary dysplasia in premature infants. Earlier we observed that aryl hydrocarbon receptor (AhR)-deficient mice are more susceptible to hyperoxic lung injury than AhR-sufficient mice, and this phenomenon was associated with a lack of expression of cytochrome P450 1A enzymes. Omeprazole, a proton pump inhibitor used in humans with gastric acid-related disorders, activates AhR in hepatocytes in vitro. However, the effects of omeprazole on AhR activation in the lungs and its impact on hyperoxia-induced reactive oxygen species (ROS) generation and inflammation are unknown. In this study, we tested the hypothesis that omeprazole attenuates hyperoxia-induced cytotoxicity, ROS generation, and expression of monocyte chemoattractant protein-1 (MCP-1) in human lung-derived H441 cells via AhR activation. Experimental groups included cells transfected with AhR small interfering RNA (siRNA). Hyperoxia resulted in significant increases in cytotoxicity, ROS generation, and MCP-1 production, which were significantly attenuated with the functional activation of AhR by omeprazole. The protective effects of omeprazole on cytotoxicity, ROS production, and MCP-1 production were lost in H441 cells whose AhR gene was silenced by AhR siRNA. These findings support the hypothesis that omeprazole protects against hyperoxic injury in vitro via AhR activation that is associated with decreased ROS generation and expression of MCP-1.

Highlights

Omeprazole treatment attenuates hyperoxia-induced ROS generation and inflammation. ► Omeprazole protects pulmonary cells against oxygen cytotoxicity. ► Omeprazole induces human CYP1A1 in H441 cells via the aryl hydrocarbon receptor. ► Aryl hydrocarbon receptor knockdown abrogates the beneficial effects of omeprazole.

Section snippets

Cell culture and treatment

H441 cells, a human lung adenocarcinoma epithelial cell line, obtained from American Type Culture Collection (Manassas, VA, USA), were grown in RPMI 1640 medium (Invitrogen, Carlsbad, CA, USA) containing 10% fetal bovine serum, 50 U/ml penicillin, and 50 U/ml streptomycin, in 95% air and 5% CO2 at 37 °C. Cells were treated with either dimethyl sulfoxide (DMSO) (Sigma–Aldrich, St. Louis, MO, USA) or 2 (OM2) or 5 μM (OM5) omeprazole (Sigma–Aldrich) for 4 h, followed by exposure to room air or

Results

In this study, we investigated the effects of omeprazole on the functional activation of AhR and its impact on hyperoxic injury in the human lung-derived H441 cells in vitro.

Discussion

This study demonstrates that the proton pump inhibitor omeprazole attenuates hyperoxic injury in vitro via the AhR, which is associated with decreased ROS generation and MCP-1 production. In human lung-derived H441 cells in vitro, omeprazole-mediated protection against hyperoxic injury correlated with the functional activation of the AhR by omeprazole compared to controls, whereas, in H441 cells whose AhR gene was silenced with AhR siRNA, the lack of omeprazole-mediated protective effects

Acknowledgments

This work was supported by grants from the Division of Neonatal–Perinatal Medicine, Texas Children's Hospital, to B.S. and the National Institutes of Health (ES-009132, HL-070921, HL-087174, and ES-019689) to B.M. We thank Esther Inman for providing technical assistance for the completion of this article.

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