In Vitro Model of Sepsis-Induced Renal Epithelial Reactive Nitrogen Species Generation

• DOI: 10.1093/toxsci/kfq058
• Publication Date: 2010-02-23T01:36:28Z
• Authors: Elina Pathak; Philip R. Mayeux
• Keywords: Reactive nitrogen species

Sepsis-induced acute kidney injury (AKI) is a complex disease characterized by generation of inducible nitric oxide synthase (iNOS)–derived reactive nitrogen species (RNS) the renal tubular epithelium. While most in vitro models sepsis use combinations lipopolysaccharide and cytokines to simulate exposure inflammatory mediators thought play role sepsis, relevance these limited. To address need for model that more closely mimics microenvironment during we developed an where mIMCD-3 (murine epithelial) cells are treated with media containing 5% serum collected from mice at 4 h after cecal ligation puncture (CLP) or sham surgery (no sepsis). After CLP serum, induction iNOS messenger RNA occurred NO was significantly increased compared sham. This increase accompanied RNS as measured oxidation 5-(and-6)-carboxy-2,7′-dichlorodihydrofluorescein diacetate (carboxy-H2DCF-DA) 2-(3,6-diamino-9H-xanthen-9-yl)-benzoic acid, methyl ester (dihydrorhodamine 123) moderate cytotoxicity similar what observed subjected CLP. Since has been shown important sepsis-induced AKI, inhibitor L-N6-(1-iminoethyl)-lysine (L-NIL) tested this model. L-NIL completely blocked generation, cytotoxicity, its effects vivo. Therefore, new exhibits many characteristics vivo, suggesting it relevant studying mechanism epithelial injury.