Increasing evidence suggests that apoptosis of tubular cells and renal inflammation mainly determine the outcome of sepsis-associated acute kidney injury (AKI). The study aim was to investigate the molecular mechanism involved in the renoprotective effects of simvastatin in endotoxin (lipopolysaccharide, LSP)-induced AKI. A sepsis model was established by intraperitoneal injection of a single non-lethal LPS dose after short-term simvastatin pretreatment. The severity of the inflammatory injury was expressed as renal damage scores (RDS). Apoptosis of tubular cells was detected by Terminal deoxynucleotidyl transferase-mediated dUTP Nick End Labeling (TUNEL assay) (apoptotic DNA fragmentation, expressed as an apoptotic index, AI) and immunohistochemical staining for cleaved caspase-3, cytochrome C, and anti-apoptotic Bcl-xL and survivin. We found that endotoxin induced severe renal inflammatory injury (RDS = 3.58 ± 0.50), whereas simvastatin dose-dependently prevented structural changes induced by LPS. Furthermore, simvastatin 40 mg/kg most profoundly attenuated tubular apoptosis, determined as a decrease of cytochrome C, caspase-3 expression, and AIs (p < 0.01 vs. LPS). Conversely, simvastatin induced a significant increase of Bcl-XL and survivin, both in the strong inverse correlations with cleaved caspase-3 and cytochrome C. Our study indicates that simvastatin has cytoprotective effects against LPS-induced tubular apoptosis, seemingly mediated by upregulation of cell-survival molecules, such as Bcl-XL and survivin, and inhibition of the mitochondrial cytochrome C and downstream caspase-3 activation.

Biomedical Research Center University Hospital Hradec Kralove 500 02 Hradec Kralove Czech Republic

Center for Biomedical Research School of Medicine University of Banja Luka 14 Save Mrkalja St 78000 Banja Luka Bosnia and Herzegovina

Department for Experimental Toxicology and Pharmacology National Poison Control Centre Military Medical Academy 11 Crnotravska St 11000 Belgrade Serbia

Department of Chemistry Faculty of Science University of Hradec Kralove Rokitanského 62 500 03 Hradec Králové Czech Republic

Department of Pharmacological Sciences Medical Faculty of the Military Medical Academy the University of Defence in Belgrade 17 Crnotravska St 11000 Belgrade Serbia

Department of Pharmacology Toxicology and Clinical Pharmacology School of Medicine University of Banja Luka 14 Save Mrkalja St 78000 Banja Luka Bosnia and Herzegovina

Institute of Pathology University Clinical Center of Republic of Srpska School of Medicine University of Banja Luka 12 Beba St 78000 Banja Luka Bosnia and Herzegovina

Special Police Unit Police Department of the City of Belgrade Ministry of Interior Trebevićka 12 A 11030 Belgrade Serbia

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