A modified approach to establish a murine model of hypoxic renal interstitial fibrosis

Modelling methods that are commonly used to establish a murine model of hypoxic renal interstitial fibrosis mainly includes 5/6 nephrectomy, unilateral ureteral obstruction and cyclosporin A (CsA)-induced renal interstitial fibrosis. The first two methods are technically challenging and unsuitable for clinical practice; thus, CsA induction is more promising. A previously introduced model of CsA-induced renal interstitial fibrosis involves the subcutaneous injection of CsA combined with a 0.01% low-sodium diet. The aim of this study was to provide a modified approach to this model by replacing the subcutaneous injection with gavage and the low-sodium diet with furosemide. From the gross morphology of kidney; the micro-specimens which were stained with haematoxylin-eosin (H&E), Masson-trichrome (Masson), periodic acid-Schiff (PAS); the renal function determination; and the expression of Vimentin protein. Our findings indicate that the combined administration of CsA every day and furosemide every other day by gavage at 80 mg/kg and 60 mg/kg, respectively, for 28 days can be used to successfully establish a murine model of renal interstitial fibrosis. Immunohistochemistry was used to show the expression of renin, the initiator of renin angiotensin aldosterone system (RAAS), while Western blotting was used to show the expression of hypoxia-inducible factor-1α (HIF-1α), a sensitive indicator of hypoxia. The expression levels of renin and HIF-1α revealed that RAAS activation and hypoxia are important mechanisms of this the model. Altogether, the data suggest that our modified approach is also an effective, alternative way to establish this model.