NBHA Reduces Acrolein-Induced Changes in ARPE-19 Cells: Possible Involvement of TGFβ

Abstract

Purpose: Acrolein, a toxic, reactive aldehyde formed metabolically and environmentally, has been implicated in the damage to and dysfunction of the retinal pigment epithelium (RPE) that accompanies age-related macular degeneration (AMD). Our purpose was to investigate the potential of acrolein to influence the release of transforming growth factor beta-2 (TGFβ2) and vascular endothelial growth factor (VEGF), to assess the ability of N-benzylhydroxylamine (NBHA) to prevent the effect of acrolein on cytokine release and reduction of viable cells, and to explore the pathway by which acrolein might be causing the increase of VEGF.

Materials and Methods: Confluent ARPE-19 cells were treated with acrolein and/or NBHA. They were also pretreated with SIS3, a specific inhibitor of SMAD 3, and ZM39923, a JAK3 inhibitor, before being treated with acrolein. Viable cells were counted; ELISA was used to measure the TGFβ2 and/or VEGF in the conditioned media.

Results: Acrolein was shown to reduce the number of viable ARPE-19 cells and to upregulate the release of the proangiogenic cytokines TGFβ2 and VEGF. Co-treatment with 200 μM NBHA significantly reduced the effects of acrolein on viable cell number and TGFβ2 release. Pretreatment of the cells with SIS3 partially blocked the action of acrolein on decreased viable cell number and VEGF upregulation, suggesting that part of the effects of acrolein are mediated by the increased levels of TGFβ and its signaling.

Conclusions: Our results suggest that the action of acrolein on the reduction of viability and VEGF increase by ARPE-19 cells is partially mediated by TGFβ2. By reducing the effects of acrolein, NBHA and SIS3 could be potential pharmacological agents in the prevention and progression of acrolein-induced damage to the RPE that relates to AMD.

KEYWORDS::

• Acrolein
• Age-related macular degeneration
• Retinal pigment epithelium
• TGFβ
• VEGF

ACKNOWLEDGMENTS

The authors wish to thank the Kronkosky Charitable Foundation, the San Antonio Neuroscience Alliance, San Antonio Life Sciences Institute, the Semp Russ Foundation of the San Antonio Area Foundation, STTM and CRSGP grants, the NEI, and the NIH Score Grant (GM08194) for their generous support of our work. We also wish to thank Brandi Betts, Ruby Ortiz, and Thomas Barron for their technical assistance; Brian Yust and Drs. Terri Krakower, Will Haskins, and Jeff Grigsby for their critical review of this manuscript.

Declaration of interest: The authors report no conflict of interest. The authors alone are responsible for the content and writing of the paper.