Contrast-induced nephropathy in an animal model: Evaluation of novel biomarkers in blood and tissue samples

Abstract

Identification of novel biomarkers of contrast-induced nephropathy (CIN) that may more accurately detect renal function changes; reflect kidney damage; assist monitoring; and elucidate pathophysiology attract considerable scientific attention nowadays. To evaluate novel biomarkers of nephrotoxicity in blood/tissue samples of a CIN model, 10 New Zealand white rabbits were divided into group 1 (n = 5; iopromide) and group 2 (n = 5; control). Blood was drawn at 0 h (immediately), 24 h and 48 h after contrast medium (CM) administration. Animals were euthanized at 48 h and kidneys were removed. Serum creatinine (sCr)/symmetric-asymmetric dimethylarginine (SDMA-ADMA) levels were measured. CM genotoxic/cytotoxic effect was investigated 48 h post-CM exposure using micronucleus assay in lymphocytes. Cytological examination was conducted using touch preparation technique (TPT). All animals in group 1 developed CIN: mean sCr levels increased by 68.2% within 48 h. Significant SDMA-ADMA level elevation was observed at 0 h and 24 h with insignificant drop at 48 h in group 1, remaining normal in group 2 at all time-points. Significant increase in bi-nucleated cells with micronuclei and micronuclei frequency was detected in group 1. Cytokinesis block proliferation index was reduced insignificantly in group 1. TPT revealed degenerative lesions/inflammation, cell degeneration, abnormal uterine tubular casts and rubella in kidneys of all animals in group 1. Group 2 presented normal cells.

Keywords: ADMA, asymmetric dimethylarginine; AKI, acute kidney injury; ANOVA, analysis of variance; ARRIVE, animal research: reporting of in vivo experiments; AVMA, American Veterinary Medical Association; Animal; Asymmetric dimethylarginine; BNMN, Bi-nucleated cells with micronuclei; CBPI, cytokinesis block proliferation index; CIN, contrast-induced nephropathy; CKD, chronic kidney disease; CM, contrast medium; Contrast media; ESI, electrospray ionization; GFR, glomerular filtration rate; Iopromide; KIM-1, kidney injury molecule-1; Kidney; LC–MS, liquid chromatography mass spectrometry; MN, micronuclei; Models; NGAL, meutrophil gelatinase–associated lipocalin; NO, nitric oxide; Nephropathy; Nephrotoxicity; OECD, Organisation for Economic Co-operation and Development; RBF, renal blood flow; ROS, reactive oxygen species; SCR, serum creatinine; SD, standard deviation; SDMA, symmetric dimethylarginine; Symmetric dimethylarginine; TPT, touch preparation technique.

Graphical abstract

Fig. 1

A microscopic picture taken during the scoring process. (A) Binucleated cell with two micronuclei. (B and C) Mononucleated lymphocyte cells with present MN. According to the methods of this assay, such MN are not considered measurable.

Fig. 2

Touch preparations (right kidney): A) Group 2: juxtaglomerular region, normal; B) Group 2: medulla, normal; C) Group 1: juxtaglomerular region, apoptosis; D) Group 1: medulla, cell degeneration.

Fig. 3

Pathophysiology of contrast induced nephropathy (CIN) - Modified from [14]. CM: contrast media, CKD: chronic kidney disease, RBF: renal blood flow, GFR: glomerular filtration rate, sCr: serum creatinine, ROS: reactive oxygen species, MN: micronuclei, BNMN: binucleated cells with micronuclei, ADMA: asymmetric dimethylarginine, SDMA: symmetric dimethylarginine.