Epigenetic memory of oxidative stress: does nephrilin exert its protective effects via Rac1?

Abstract

Aim

Nephrilin peptide, a designed inhibitor of Rictor complex (mTORC2), exerts pleiotropic protective effects in metabolic, xenobiotic and traumatic stress models. Stress can generate enduring epigenetic changes in gene function. In this work we examine the possibility that nephrilin treatment protects against acute and enduring global changes in oxidative metabolism, with a focus on the Rictor-complex-mediated activation of Rac1, a subunit of NADPH oxidase (Nox) via PKCs, Prex1 and p66shc.

Methods

Given the wide range of animal models in which nephrilin peptide has previously demonstrated effectiveness in vivo, we chose three different experimental systems for this investigation: dermal fibroblasts, renal proximal tubule epithelial cells (PTECs), and kidney tissue and urine from an animal model of burn trauma in which nephrilin was previously shown to prevent loss of kidney function.

Results

(1) Nephrilin protects dermal fibroblasts from loss of viability and collagen synthesis after ultraviolet A (UV-A) or H₂O₂ insult. (2) Nephrilin reduces reactive oxygen species (ROS) formation by H₂O₂–treated (PTECs) with or without nicotine pretreatment. Using RNA arrays and pathway analysis we demonstrate that nicotine and H₂O₂-treated PTECs specifically induced Rac1 gene networks in these cells. (3) Using kidney tissue and urine from the burn trauma model we demonstrate significant elevations of [a] 8-aminoprostane in urine; [b] kidney tissue histone modification and DNA methylation; and [c] post-transcriptional phosphorylation events consistent with Rac1 activation in kidney tissue.

Conclusion

Nephrilin protects against oxidative stress, possibly by modulating the activation of Rac1.

Keywords:

• nephrilin
• epigenetic
• Rac1
• burn injury
• Rictor
• 8-isoprostane

Acknowledgments

We thank Dr. Celeste Finnerty and Dr. Amina ElAyadi for their generous gift of frozen rat tissues used in this study. This work was supported in part by the Department of Pediatrics at the University of Mississippi Medical Center and the Bower Foundation.

Disclosure

The authors report no conflicts of interest in this work.