Abstract
Recently, there has been an increase in the use of several nephrotoxicity biomarkers in preclinical experiments. In addition, it has been indicated that the result may have been influenced by secondary factors, such as sample storage condition or storage period. In this study, we have assessed the variation in urinary nephrotoxicity biomarkers as a result of urine storage conditions and storage period of the urine. Urine was sampled from specific pathogen-free Sprague-Dawley rats (19 weeks old), which were housed individually in hanged stainless steel wire mesh cages. Urine was stored at 20oC, at 4oC, or at −70oC after sampling. The levels of the biomarkers such as beta-2 microglobulin (B2M), cystatin-C (Cys-C), N-acetyl-β-D-glucosaminidase (NAG), micro albumin (MA), micro protein (MP) were measured at 6, 24, 48 and 144 hr after sampling. The B2M level was significantly decreased at 6, 24, 48, and 144 hr compared to 0 hr at −70oC (p < 0.05, p < 0.01, p < 0.05, and p < 0.05, respectively) and 24 and 144 hr at 20oC (p < 0.01, p < 0.01, respectively). The Cys-C level was significantly decreased at 144 hr compared to 0 hr at 4oC (p < 0.01), at 20oC (p < 0.05) and at 70oC (p < 0.01). MP and MA levels were not different for 144 hr in all storage conditions. Taken together, B2M and Cys-C levels were modulated by storage temperature and period. For the enhancement of test accuracy, it is suggested that strict protocols be established for samples to minimize the effects of the storage conditions on the detected levels of biomarkers.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
References
Dieterle, F., Perentes, E., Cordier, A., Roth, D.R., Verdes, P., Grenet, O., Pantano, S., Moulin, P., Wahl, D., Mahl, A., End, P., Staedtler, F., Legay, F., Carl, K., Laurie, D., Chibout, S.D., Vonderscher, J. and Maurer, G. (2010) Urinary clusterin, cystatin C, beta2-microglobulin and total protein as markers to detect drug-induced kidney injury. Nat. Biotechnol., 28, 463–469.
Vaidya, V.S., Waikar, S.S., Ferguson, M.A., Collings, F.B., Sunderland, K., Gioules, C., Bradwin, G., Matsouaka, R., Betensky, R.A., Curhan, G.C. and Bonventre, J.V. (2008) Urinary biomarkers for sensitive and specific detection of acute kidney injury in humans. Clin. Transl. Sci., 1, 200–208.
Belcher, J.M., Edelstein, C.L. and Parikh, C.R. (2011) Clinical applications of biomarkers for acute kidney injury. Am. J. Kidney Dis., 57, 930–940.
Waikar, S.S. and Bonventre, J.V. (2008) Biomarkers for the diagnosis of acute kidney injury. Nephron Clin. Pract., 109, c192–197.
Kim, S.Y. and Moon, A. (2012) Drug-induced nephrotoxicity and its biomarkers. Biomol. Ther. (Seoul), 20, 268–272.
Ozer, J.S., Dieterle, F., Troth, S., Perentes, E., Cordier, A., Verdes, P., Staedtler, F., Mahl, A., Grenet, O., Roth, D.R., Wahl, D., Legay, F., Holder, D., Erdos, Z., Vlasakova, K., Jin, H., Yu, Y., Muniappa, N., Forest, T., Clouse, H.K., Reynolds, S., Bailey, W.J., Thudium, D.T., Topper, M.J., Skopek, T.R., Sina, J.F., Glaab, W.E., Vonderscher, J., Maurer, G., Chibout, S.D., Sistare, F.D. and Gerhold, D.L. (2010) A panel of urinary biomarkers to monitor reversibility of renal injury and a serum marker with improved potential to assess renal function. Nat. Biotechnol., 28, 486–494.
Yu, Y., Jin, H., Holder, D., Ozer, J.S., Villarreal, S., Shughrue, P., Shi, S., Figueroa, D.J., Clouse, H., Su, M., Muniappa, N., Troth, S.P., Bailey, W., Seng, J., Aslamkhan, A.G., Thudium, D., Sistare, F.D. and Gerhold, D.L. (2010) Urinary biomarkers trefoil factor 3 and albumin enable early detection of kidney tubular injury. Nat. Biotechnol., 28, 470–477.
Dieterle, F., Sistare, F., Goodsaid, F., Papaluca, M., Ozer, J.S., Webb, C.P., Baer, W., Senagore, A., Schipper, M.J., Vonderscher, J., Sultana, S., Gerhold, D.L., Phillips, J.A., Maurer, G., Carl, K., Laurie, D., Harpur, E., Sonee, M., Ennulat, D., Holder, D., Andrews-Cleavenger, D., Gu, Y.Z., Thompson, K.L., Goering, P.L., Vidal, J.M., Abadie, E., Maciulaitis, R., Jacobson-Kram, D., Defelice, A.F., Hausner, E.A., Blank, M., Thompson, A., Harlow, P., Throckmorton, D., Xiao, S., Xu, N., Taylor, W., Vamvakas, S., Flamion, B., Lima, B.S., Kasper, P., Pasanen, M., Prasad, K., Troth, S., Bounous, D., Robinson-Gravatt, D., Betton, G., Davis, M.A., Akunda, J., McDuffie, J.E., Suter, L., Obert, L., Guffroy, M., Pinches, M., Jayadev, S., Blomme, E.A., Beushausen, S.A., Barlow, V.G., Collins, N., Waring, J., Honor, D., Snook, S., Lee, J., Rossi, P., Walker, E. and Mattes, W. (2010) Renal biomarker qualification submission: a dialog between the FDA-EMEA and Predictive Safety Testing Consortium. Nat. Biotechnol., 28, 455–462.
Mall, C., Rocke, D.M., Durbin-Johnson, B. and Weiss, R.H. (2013) Stability of miRNA in human urine supports its biomarker potential. Biomarkers Med., 7, 623–631.
Parikh, C.R., Butrymowicz, I., Yu, A., Chinchilli, V.M., Park, M., Hsu, C.Y., Reeves, W.B., Devarajan, P., Kimmel, P.L., Siew, E.D. and Liu, K.D. (2014) Urine stability studies for novel biomarkers of acute kidney injury. Am. J. Kidney Dis., 63, 567–572.
Miki, K. and Sudo, A. (1998) Effect of urine pH, storage time, and temperature on stability of catecholamines, cortisol, and creatinine. Clin. Chem., 44, 1759–1762.
Jung, K., Pergande, M., Schreiber, G. and Schröder, K. (1983) Stability of enzymes in urine at 37 degrees C. Clin. Chim. Acta, 131, 185–191.
Hoffmann, D., Fuchs, T.C., Henzler, T., Matheis, K.A., Herget, T., Dekant, W., Hewitt, P. and Mally, A. (2010) Evaluation of a urinary kidney biomarker panel in rat models of acute and subchronic nephrotoxicity. Toxicology, 277, 49–58.
Schentag, J.J., Sutfin, T.A., Plaut, M.E. and Jusko, W.J. (1978) Early detection of aminoglycoside nephrotoxicity with urinary beta-2-microglobulin. J. Med., 9, 201–210.
Davey, P.G. and Gosling, P. (1982) beta 2-Microglobulin instability in pathological urine. Clin. Chem., 28, 1330–1333.
Herget-Rosenthal, S., Feldkamp, T., Volbracht, L. and Kribben, A. (2004) Measurement of urinary cystatin C by particleenhanced nephelometric immunoassay: precision, interferences, stability and reference range. Ann. Clin. Biochem., 41, 111–118.
Park, M.Y., Choi, S.J., Kim, J.K., Hwang, S.D. and Lee, Y.W. (2013) Urinary cystatin C levels as a diagnostic and prognostic biomarker in patients with acute kidney injury. Nephrology (Carlton), 18, 256–262.
Morita, A., Numata, Y., Kosugi, Y., Noto, A., Takeuchi, N. and Uchida, K. (1998) Stabilities of N-acetyl-beta-D-glucos-aminidase (NAG) isoenzymes in urine: advantage of NAG isoenzyme B measurement in clinical applications. Clin. Chim. Acta, 278, 35–43.
Price, R.G. (1982) Urinary enzymes, nephrotoxicity and renal disease. Toxicology, 23, 99–134.
Usuda, K., Kono, K., Dote, T., Nishiura, K., Miyata, K., Nishiura, H., Shimahara, M. and Sugimoto, K. (1998) Urinary biomarkers monitoring for experimental fluoride nephrotoxicity. Arch. Toxicol., 72, 104–109.
Author information
Authors and Affiliations
Corresponding authors
Rights and permissions
This is an Open-Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
About this article
Cite this article
Lee, JM., Han, YH., Choi, SJ. et al. Variation of Nephrotoxicity Biomarkers by Urinary Storage Condition in Rats. Toxicol Res. 30, 305–309 (2014). https://doi.org/10.5487/TR.2014.30.4.305
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.5487/TR.2014.30.4.305