Neutrophil gelatinase-associated lipocalin, a sensitive urinary biomarker of acute kidney injury in dogs receiving gentamicin

Abstract

-A sensitive urinary biomarker for acute kidney injury (AKI) was investigated in beagle dogs with nephrotoxicity induced by gentamicin. Gentamicin sulphate at 25 or 50 mg/kg was injected (s.c.) for 9 days, and conventional urinalysis, ELISA assay of neutrophil gelatinase-associated lipocal (NGAL) in urine, blood chemistry, and pathological examinations were performed. The dog given gentamicin at 25 mg/kg only showed slight deposition of lysosomal granules in the proximal tubular epithelium of the kidneys without any other significant changes even though urinary NGAL was elevated on Day 10 (day of necropsy). In the dog receiving gentamicin at 50 mg/kg, increases in urinary NGAL were observed on Days 3 and 5, and absence of urination, marked increases in serum urea nitrogen and creatinine, enlargement and discoloration of the kidneys with marked necrosis, and swelling of proximal epithelium were observed. In conclusion, urinary NGAL is considered to be a candidate as a sensitive predictable biomarker of AKI in the gentamicin-induced nephrotoxicity model in dogs.Key words: NGAL, Dogs, Acute kidney injury, Urinary biomarker, Gentamicin Correspondence: Kiyonori Kai (E-mail: kai.kiyonori.xb@daiichisankyo.co.jp) LetterThe Journal of Toxicological Sciences (J. Toxicol. Sci.) Vol.38, No.2, 269-277, 2013 Vol. 38 No. 2 269 addition to these biomarkers, neutrophil gelatinase-associated lipocal (NGAL), a member of the lipocalin superfamily, has been proposed as a candidate for a sensitive urinary biomarker to detect AKI Mori, 2005;Mori and Nakao, 2007). Unfortunately, in part due to the lack of commercially available assays for non-rodents, limited data comparing the performance of the exploratory and approved markers are publicly available so far.In this study, we selected gentamicin, a well-known nephrotoxic agent that induces AKI in experimental animals and humans, and injected it subcutaneously to beagle dogs for 9 days and evaluated urinary NGAL using a commercial ELISA kit as a sensitive urinary biomarker in comparison with conventional blood and urinary toxicity makers and pathological examinations. MATERIALS AND METHODS Test articleELTACIN injection, an injectable product of gentamicin sulfate was purchased from FujiPharma (Shizuoka, Japan). Animals and housing conditionsTwo male beagle dogs were purchased from Narc Corporation (Chiba, Japan) for gentamicin treatment, and 6 males and 6 females were additionally obtained from Marshall BioResources, Japan (Ibaraki, Japan) for collection background data of urinalysis. The dogs were individually housed in stainless steel cages (W93.5 cm × W80 cm × H78.5 cm) in an air-conditioned room (temperature, 18 to 28°C; relative humidity, 30 to 70%). A light/dark cycle of 12 hr and ventilation rate of 10 to 20 air changes/ hr were used in the animal rooms. Two hundred and twenty grams of basal diet (Certified Canine Diet 5007, PMI Nutrition International, Inc., St. Louis, MO, USA) was given to each dog in the morning and tap water was available ad libitum. All experimen...

Usefulness of urinary biomarkers for nephrotoxicity in cynomolgus monkeys treated with gentamicin, cisplatin, and puromycin aminonucleoside

“…Sustained increase in urinary NGAL was reported to be as long as 5 days post damage in humans. The increase in urinary NGAL is considered to be due to an increase in NGAL synthesis and failure of complete reabsorption of NGAL in the proximal tubular epithelia (Kai et al, 2013;Alge and Arthur, 2015). The results in the present study suggested that NGAL could be the most useful BM for the detection of earlier damages of the proximal and/or distal tubules among all urinary BMs and enzymes in cynomolgus monkey.…”

“…The usefulness of NGAL to detect the damage of the renal tubules has been reported in rats (Han et al, 2012;Phillips et al, 2016) and dogs (Kai et al, 2013;Zhou et al, 2014). After renal ischemia or nephrotoxicity, intrarenal NAGL synthesis is dramatically upregulated at protein levels.…”

Correlation of Urine and Serum Biomarkers with Renal Damage and Survival in Dogs with Naturally Occurring Proteinuric Chronic Kidney Disease

“…Urine immunoglobulin G (uIgG) and urine retinol binding protein (uRBP) were increased in dogs with primary CKD7 and those with renal dysfunction secondary to various systemic diseases including pyometra, babesiosis, and snake envenomation 7, 8, 9, 10, 11, 12, 13, 14, 15. Urine, plasma, and serum neutrophil gelatinase‐associated lipocalin (NGAL) and urine N‐acetyl‐β‐D‐glucosaminidase (uNAG), a renal tubular enzyme, are tubular markers increased in both acute and CKD in dogs 7, 11, 12, 15, 16, 17, 18, 19, 20, 21, 22. Neutrophil gelatinase‐associated lipocalin originates not only in the renal tubules but also from neutrophil granules and many other organs 23.…”

“…Neutrophil gelatinase‐associated lipocalin originates not only in the renal tubules but also from neutrophil granules and many other organs 23. Novel urine biomarkers are not regularly used as diagnostic tools for evaluation of renal disease in veterinary medicine, and few veterinary studies correlate biomarkers with histologically proven renal damage7, 12, 13, 18 and case outcome 4…”

Urinary biomarkers of renal function in dogs and cats: a review

“…The significant increase of NGAL in urine was detected as early as 12 h after surgery, much earlier than the increase in serum creatinine. Urinary NGAL was recommended as a sensitive and specific biomarker for the detection of AKI, for prediction of nephrotoxicity, and to screen patients at risk for AKI (Table 1) (Lee et al 2012;Kai et al 2013;Hsu et al 2014a;Zhou et al 2014). NGAL was found to be a more sensitive biomarker for the detection of gentamicin-induced renal proximal tubular toxicity than NAG (Zhou et al 2014).…”