Evaluation of Neutrophil Gelatinase-Associated Lipocalin as a Predictor of Glomerular Filtration Rate and Amikacin Clearance During Early Rat Endotoxemia: Comparison with Traditional Endogenous and Exogenous Biomarkers

  • Šárka Studená
  • Eva Doleželová
  • Jolana Cermanová
  • Alena Prašnická
  • Drahomíra Springer
  • Stanislav Mičuda
  • Jaroslav ChládekEmail author
Original Research Article


Background and Objectives

Renal elimination of amikacin and other aminoglycosides is slowed down in sepsis-induced acute kidney injury increasing the risk of adverse effects. Since neutrophil gelatinase-associated lipocalin (NGAL) and aminoglycosides share the mechanisms for renal excretion, the predictive power of NGAL was examined towards the changes in amikacin pharmacokinetics during early endotoxemia in anesthetized Wistar rats.


Endogenous biomarkers of inflammation and acute kidney injury were assessed including NGAL in saline-injected controls and two groups of rats challenged with an intravenous injection of bacterial lipopolysaccharide (5 mg/kg)—a fluid-resuscitated group (LPS) and a fluid-resuscitated group infused intravenously with 8 μg/kg/h terlipressin (LPS-T). Sinistrin and amikacin were infused to measure glomerular filtration rate (GFR) and amikacin clearance (CLam). The investigations included blood gas analysis, chemistry and hematology tests and assessment of urine output, creatinine clearance (CLcr) and sinistrin clearance (CLsini).


Within 3 h of injection, systemic and renal inflammatory responses were induced by lipopolysaccharide. Gene and protein expression of NGAL was increased in the kidneys and the concentrations of NGAL in the plasma (pNGAL) and urine rose 4- to 38-fold (P < 0.01). The decreases in CLam and the GFR markers (CLcr, CLsini) were proportional, reflecting the extent to which endotoxemia impaired the major elimination mechanism for the drug. Terlipressin attenuated lipopolysaccharide-induced renal dysfunction (urine output, CLcr, CLsini) and accelerated CLam. The pNGAL showed a strong association with the CLsini (rs = − 0.77, P < 0.0005). Concerning prediction of CLam, pNGAL was comparable to CLcr (mean error − 24%) and inferior to CLsini (mean error − 6.4%), while the measurement of NGAL in urine gave unsatisfactory results.


During early endotoxemia in the rat, pNGAL has a moderate predictive ability towards CLam. Clinical studies should verify whether pNGAL can support individualized dosing of aminoglycosides to septic patients.


Author Contributions

All authors contributed to the study conception and design. Material preparation, data collection and analysis were performed by ŠS, ED, JC, AP, DS and SM. The first draft of the manuscript was written by JC and all authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.

Compliance with Ethical Standards


This study was supported by the grant from the Grant Agency of Charles University Progres Q40.

Conflict of interest

The authors declare no conflicts of interest.

Ethical approval

The study was approved by the Animal Welfare Committee at the Charles University in Prague, Faculty of Medicine in Hradec Kralove. The animals received humane care according to the ethical standards of Directive 86/609/EEC, “European Convention for the Protection of Vertebrate Animals Used for Experimental and other Scientific Purposes” (1986), and the “Guiding Principles in the Use of Animals in Toxicology” (1989).

Supplementary material

13318_2019_579_MOESM1_ESM.pdf (83 kb)
Supplementary material 1 (PDF 82 kb)


  1. 1.
    Avent ML, Rogers BA, Cheng AC, Paterson DL. Current use of aminoglycosides: indications, pharmacokinetics and monitoring for toxicity. Intern Med J. 2011;41:441–9.CrossRefGoogle Scholar
  2. 2.
    Pagkalis S, Mantadakis E, Mavros MN, Ammari C, Falagas ME. Pharmacological considerations for the proper clinical use of aminoglycosides. Drugs. 2011;71:2277–94.CrossRefGoogle Scholar
  3. 3.
    Nagai J, Takano M. Molecular aspects of renal handling of aminoglycosides and strategies for preventing the nephrotoxicity. Drug Metab Pharmacokinet. 2004;19:159–70.CrossRefGoogle Scholar
  4. 4.
    Marsot A, Guilhaumou R, Riff C, Blin O. Amikacin in critically ill patients: a review of population pharmacokinetic studies. Clin Pharmacokinet. 2017;56:127–38.CrossRefGoogle Scholar
  5. 5.
    Seegmiller JC, Eckfeldt JH, Lieske JC. Challenges in measuring glomerular filtration rate: a clinical laboratory perspective. Adv Chronic Kidney Dis. 2018;25:84–92.CrossRefGoogle Scholar
  6. 6.
    Borregaard N, Cowland JB. Neutrophil gelatinase-associated lipocalin, a siderophore-binding eukaryotic protein. Biometals. 2006;19:211–5.CrossRefGoogle Scholar
  7. 7.
    Mori K, Nakao K. Neutrophil gelatinase-associated lipocalin as the real-time indicator of active kidney damage. Kidney Int. 2007;71:967–70.CrossRefGoogle Scholar
  8. 8.
    Schrezenmeier EV, Barasch J, Budde K, Westhoff T, Schmidt-Ott KM. Biomarkers in acute kidney injury –pathophysiological basis and clinical performance. Acta Physiol. 2017;219:554–72.CrossRefGoogle Scholar
  9. 9.
    Beker BM, Corleto MG, Fieiras C, Musso CG. Novel acute kidney injury biomarkers: their characteristics, utility and concerns. Int Urol Nephrol. 2018;50(4):705–13.CrossRefGoogle Scholar
  10. 10.
    Downes KJ, Dong M, Fukuda T, Clancy JP, Haffner C, Bennett MR, et al. Urinary kidney injury biomarkers and tobramycin clearance among children and young adults with cystic fibrosis: a population pharmacokinetic analysis. J Antimicrob Chemother. 2017;72:254–60.CrossRefGoogle Scholar
  11. 11.
    Morelli A, Ertmer C, Rehberg S, Lange M, Orecchioni A, Cecchini V, et al. Continuous terlipressin versus vasopressin infusion in septic shock (TERLIVAP): a randomized, controlled pilot study. Crit Care. 2009;13:R130.CrossRefGoogle Scholar
  12. 12.
    Qiu X, Huang Y, Xu J, Qiu H, Yang Y. Effects of terlipressin on microcirculation of small bowel mesentery in rats with endotoxic shock. J Surg Res. 2014;188:503–9.CrossRefGoogle Scholar
  13. 13.
    Pill J, Issaeva O, Woderer S, Sadick M, Kränzlin B, Fiedler F, et al. Pharmacological profile and toxicity of fluorescein-labelled sinistrin, a novel marker for GFR measurements. Naunyn Schmiedebergs Arch Pharmacol. 2006;373:204–11.CrossRefGoogle Scholar
  14. 14.
    Woitzik J, Abromeit N, Schaefer F. Measurement of nitric oxide metabolites in brain microdialysates by a sensitive fluorometric high-performance liquid chromatography assay. Anal Biochem. 2001;289:10–7.CrossRefGoogle Scholar
  15. 15.
    Kadova Z, Dolezelova E, Cermanova J, Hroch M, Laho T, Muchova L, et al. IL-1 receptor blockade alleviates endotoxin-mediated impairment of renal drug excretory functions in rats. Am J Physiol Renal Physiol. 2015;308:F388–99.CrossRefGoogle Scholar
  16. 16.
    Kim H, Hur M, Lee S, Marino R, Magrini L, Cardelli P, et al. Proenkephalin, neutrophil gelatinase-associated lipocalin, and estimated glomerular filtration rates in patients with sepsis. Ann Lab Med. 2017;37:388–97.CrossRefGoogle Scholar
  17. 17.
    Otto GP, Hurtado-Oliveros J, Chung HY, Knoll K, Neumann T, Müller HJ, et al. Plasma neutrophil gelatinase-associated lipocalin is primarily related to inflammation during sepsis: a translational approach. PLoS One. 2015;10:e0124429.CrossRefGoogle Scholar
  18. 18.
    Kees MG, Hilpert JW, Gnewuch C, Kees F, Voegeler S. Clearance of vancomycin during continuous infusion in intensive care unit patients: correlation with measured and estimated creatinine clearance and serum cystatin C. Int J Antimicrob Agents. 2010;36:545–8.CrossRefGoogle Scholar
  19. 19.
    Kees MG, Minichmayr IK, Moritz S, Beck S, Wicha SG, Kees F, et al. Population pharmacokinetics of meropenem during continuous infusion in surgical ICU patients. J Clin Pharmacol. 2016;56:307–15.CrossRefGoogle Scholar
  20. 20.
    Han M, Li Y, Liu M, Li Y, Cong B. Renal neutrophil gelatinase associated lipocalin expression in lipopolysaccharide-induced acute kidney injury in the rat. BMC Nephrol. 2012;13:25.CrossRefGoogle Scholar
  21. 21.
    Arulkumaran N, Sixma ML, Jentho E, Ceravola E, Bass PS, Kellum JA, et al. Sequential analysis of a panel of biomarkers and pathologic findings in a resuscitated rat model of sepsis and recovery. Crit Care Med. 2017;45:e821–30.CrossRefGoogle Scholar
  22. 22.
    Otto GP, Busch M, Sossdorf M, Claus RA. Impact of sepsis-associated cytokine storm on plasma NGAL during acute kidney injury in a model of polymicrobial sepsis. Crit Care. 2013;17:419.CrossRefGoogle Scholar
  23. 23.
    Axelsson L, Bergenfeldt M, Ohlsson K. Studies of the release and turnover of a human neutrophil lipocalin. Scand J Clin Lab Investig. 1995;55:577–88.CrossRefGoogle Scholar
  24. 24.
    Mori K, Lee HT, Rapoport D, Drexler IR, Foster K, Yang J, et al. Endocytic delivery of lipocalin-siderophore-iron complex rescues the kidney from ischemia-reperfusion injury. J Clin Investig. 2005;115:610–21.CrossRefGoogle Scholar
  25. 25.
    Mårtensson J, Bell M, Xu S, Bottai M, Ravn B, Venge P, et al. Association of plasma neutrophil gelatinase-associated lipocalin (NGAL) with sepsis and acute kidney dysfunction. Biomarkers. 2013;18:349–56.CrossRefGoogle Scholar
  26. 26.
    Tardif D, Beauchamp D, Bergeron MG. Influence of endotoxin on the intracortical accumulation kinetics of gentamicin in rats. Antimicrob Agents Chemother. 1990;34:576–80.CrossRefGoogle Scholar
  27. 27.
    Roberts JA. Pharmacokinetic issues for antibiotics in the critically ill patient. Crit Care Med. 2009;37:840–51.CrossRefGoogle Scholar
  28. 28.
    Rittirsch D, Hoesel LM, Ward PA. The disconnect between animal models of sepsis and human sepsis. J Leukoc Biol. 2007;81:137–43.CrossRefGoogle Scholar

Copyright information

© Springer Nature Switzerland AG 2019

Authors and Affiliations

  1. 1.Department of PharmacologyCharles University, Faculty of Medicine Hradec KrálovéHradec KrálovéCzech Republic
  2. 2.Department of Biological and Medical SciencesCharles University, Faculty of PharmacyHradec KrálovéCzech Republic
  3. 3.Institute of Medical Biochemistry and Laboratory Diagnostics, 1st Faculty of MedicineCharles University in Prague and General University Hospital in PraguePraha 2Czech Republic

Personalised recommendations