Podocyte NADPH Oxidase 5 Promotes Renal Inflammation Regulated by the Toll-Like Receptor Pathway

Aims: Oxidative stress associated with a proinflammatory state occurs in endothelial dysfunction, hypertension, chronic kidney disease, and diabetes. The NADPH oxidase (Nox) family of reactive oxygen species (ROS) generating enzymes is implicated in these processes, yet little information regarding the role of Nox5 is available. Our aim was to investigate the role of Nox5 in promoting renal inflammation and identify mechanisms regulating its activity. Results: Mice with podocyte-specific Nox5 (Nox5(pod+)) expression demonstrated greater glomerular inflammation and increased expression of Toll-like receptors (TLRs) and proinflammatory cytokines. In a lipopolysaccharide (LPS) model of acute kidney injury, Nox5(pod+) and control littermates exhibited increased TLR and Nox1 expression. Compared with control littermates, Nox5(pod+) animals developed greater glomerular inflammation and ROS production. Immortalized human podocytes (hPODs) incubated with LPS demonstrated TLR induction, increased Nox5 expression, and enhanced ROS production. Inhibition of interleukin-1 receptor-associated kinases (IRAK)-1 and -4 that lie downstream of TLR inhibited LPS-induced ROS production. Interaction between IRAK1 and Nox5 was confirmed by coimmunoprecipitation. Furthermore, LPS treatment of hPODs resulted in phosphorylation of threonine residue(s) in Nox5 that was attenuated by an IRAK1/4 inhibitor. Innovation and Conclusion: These results are the first to demonstrate that Nox5 is a downstream target of the TLR pathway and that Nox5-derived ROS may be modulated by IRAK1/4 activity. Nox5-derived ROS in podocytes can promote a proinflammatory state in the kidney via induction of cytokine expression and upregulation of TLRs leading to a feed-forward loop in which TLR activation enhances Nox5-mediated ROS production.