Abstract
Objective
To find new protein biomarkers for the detection and evaluation of liver injury and to analyze the relationship between such proteins and disease progression in concanavalin A (Con A)-induced hepatitis.
Methods
Twenty-five mice were randomly divided into five groups: an untreated group, a control group injected with phosphate buffered saline (PBS), and groups with Con A-induced hepatitis evaluated at 1, 3 and 6 h. Two-dimensional gel electrophoresis (2-DE) and mass spectrometry (MS) were used to identify differences in protein expression among groups. Quantitative real-time polymerase chain reaction (qRT-PCR) was performed to verify the results.
Results
In mice with Con A-induced hepatitis, expression levels of four proteins were increased: RIKEN, fructose bisphosphatase 1 (fbp1), ketohexokinase (khk), and Chain A of class pi glutathione S-transferase. Changes in fbp1 and khk were confirmed by qRT-PCR.
Conclusion
Levels of two proteins, fbp1 and khk, are clearly up-regulated in mice with Con A-induced hepatitis.
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References
Bodzon-Kulakowska, A., Bierczynska-Krzysik, A., Dylag, T., Drabik, A., Suder, P., Noga, M., Jarzebinska, J., Silberring, J., 2007. Methods for samples preparation in proteomic research. J. Chromatogr. B, 849(1–2):1–31. [doi: 10.1016/j.jchromb.2006.10.040]
Bonthron, D.T., Brady, N., Donaldson, I.A., Steinmann, B., 1994. Molecular basis of essential fructosuria: molecular cloning and mutational analysis of human ketohexokinase (fructokinase). Hum. Mol. Genet., 3(9):1627–1631. [doi: 10.1093/hmg/3.9.1627]
Cuesta, E., Boada, J., Calafell, R., Perales, J.C., Roig, T., Bermudez, J., 2006. Fructose 1,6-bisphosphate prevented endotoxemia, macrophage activation, and liver injury induced by D-galactosamine in rats. Crit. Care Med., 34(3):807–814. [doi:10.1097/01.CCM.0000202016.60856.03]
Dominguez, D.C., Lopes, R., Torres, M.L., 2007. Proteomics technology. Clin. Lab. Sci., 20(4):239–244.
Duncan, R., Bazar, L., Michelotti, G., Tomonaga, T., Krutzsch, H., Avigan, M., Levens, D., 1994. A sequence-specific, single-strand binding protein activates the far upstream element of c-myc and defines a new DNA-binding motif. Genes Dev., 8(4):465–480. [doi:10.1101/gad.8.4.465]
Farago, N., Kocsis, G.F., Feher, L.Z., Csont, T., Hackler, L.Jr., Varga-Orvos, Z., Csonka, C., Kelemen, J.Z., Ferdinandy, P., Puskas, L.G., 2008. Gene and protein expression changes in response to normoxic perfusion in mouse hearts. J. Pharmacol. Toxicol. Meth., 57(2):145–154. [doi:10.1016/j.vascn.2008.01.001]
García-Sáez, I., Párraga, A., Phillips, M.F., Mantle, T.J., Coll, M., 1994. Molecular structure at 1.8 A of mouse liver class pi glutathione S-transferase complexed with S-(p-nitrobenzyl) glutathione and other inhibitors. J. Mol. Biol., 237(3):298–314. [doi:10.1006/jmbi.1994.1232]
He, Q.Y., Lau, G.K., Zhou, Y., Yuen, S.T., Lin, M.C., Kung, H.F., Chiu, J.F., 2003. Serum biomarkers of hepatitis B virus infected liver inflammation: a proteomic study. Proteomics, 3(5):666–674. [doi:10.1002/pmic.200300394]
Henkel, C., Roderfeld, M., Weiskirchen, R., Berres, M.L., Hillebrandt, S., Lammert, F., Meyer, H.E., Stühler, K., Graf, J., Roeb, E., 2006. Changes of the hepatic proteome in murine models for toxically induced fibrogenesis and sclerosing cholangitis. Proteomics., 6(24):6538–6548. [doi:10.1002/pmic.200600580]
Horecker, B.L., Melloni, E., Pontremoli, S., 1975. Fructose 1,6-bisphosphatase: properties of the neutral enzyme and its modification by proteolytic enzymes. Adv. Enzymol. Relat. Areas Mol. Biol., 42:193–226. [doi:10.1002/9780470122877.ch4]
Hwa, J.S., Kim, H.J., Goo, B.M., Park, H.J., Kim, C.W., Chung, K.H., Park, H.C., Chang, S.H., Kim, Y.W., Kim, D.R., et al., 2006. The expression of ketohexokinase is diminished in human clear cell type of renal cell carcinoma. Proteomics, 6(3):1077–1084. [doi:10.1002/pmic.200401345]
Kelso, A., Gough, N.M., 1987. Expression of Hemopoietic Growth Factor Genes in Murine T Lymphocytes. In: Webb, D.R., Goeddel, D.V. (Eds.), Lymphokines. Vol. 13, Academic Press Inc., London, p.209–238.
McBurney, R.N., Hines, W.M., von Tungeln, L.S., Schnackenberg, L.K., Beger, R.D., Moland, C.L., Han, T., Fuscoe, J.C., Chang, C.W., Chen, J.J., et al., 2009. The liver toxicity biomarker study: phase I design and preliminary results. Toxicol. Pathol., 37(1):52–64. [doi:10.1177/0192623308329287]
O’Farrell, P.H., 1975. High resolution two-dimensional electrophoresis of proteins. J. Biol. Chem., 250(10): 4007–4021.
Párraga, A., Garcá-Sáez, I., Walsh, S.B., Mantle, T.J., Coll, M., 1998. The three-dimensional structure of a class-pi glutathione S-transferase complexed with glutathione: the active-site hydration provides insights into the reaction mechanism. Biochem. J., 333(Pt 3):811–816.
Puppala, S., Dodd, G.D., Fowler, S., Arya, R., Schneider, J., Farook, V.S., Granato, R., Dyer, T.D., Almasy, L., Jenkinson, C.P., et al., 2006. A genomewide search finds major susceptibility loci for gallbladder disease on chromosome 1 in Mexican Americans. Am. J. Hum. Genet., 78(3):377–392. [doi:10.1086/500274]
Tiegs, C., Hentschel, J., Wendel, A., 1992. A cell-dependent experimental liver injury in mice inducible by concanavalin A. J. Clin. Invest., 90(1):196–203. [doi:10.1172/JCI115836]
van Belle, W., Ånensen, N., Haaland, I., Bruserud, Ø., Høgda, K.A., Gjertsen, B.T., 2006. Correlation analysis of two-dimensional gel electrophoretic protein patterns and biological variables. BMC Bioinf., 7(1):198. [doi:10.1186/1471-2105-7-198]
Wang, T., Foker, J.E., Tsai, M.Y., 1980. The shift of an increase in phosphofructokinase activity from protein synthesis-dependent to -independent mode during concanavalin A induced lymphocyte proliferation. Biochem. Biophys. Res. Commun., 95(1):13–19. [doi:10.1016/0006-291X(80)90697-X]
Wilm, M., Shevchenko, A., Houthaeve, T., Breit, S., Schweigerer, L., Fotsis, T., Mann, M., 1996. Femtomole sequencing of proteins from polyacrylamide gels by nano-electrospray mass spectrometry. Nature, 379(6564): 466–469. [doi:10.1038/379466a0]
Yu, M., Wang, X.X., Zhang, F.R., Shang, Y.P., Du, Y.X., Chen, H.J., Chen, J.Z., 2007. Proteomic analysis of the serum in patients with idiopathic pulmonary arterial hypertension. J. Zhejiang Univ.-Sci. B, 8(4):221–227. [doi: 10.1631/jzus.2007.B0221]
Yuan, J.S., Reed, A., Chen, F., Stewart, C.N.Jr., 2006. Statistical analysis of real-time PCR data. BMC Bioinf., 7(1):85. [doi:10.1186/1471-2105-7-85]
Zhang, A.S., Xiong, S., Tsukamoto, H., Enns, C.A., 2004. Localization of iron metabolism-related mRNAs in rat liver indicate that HFE is expressed predominantly in hepatocytes. Blood, 103(4):1509–1514. [doi:10.1182/blood-2003-07-2378]
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Project (No. 20082X10002-007) supported by the National Natural Science Foundation of China
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Tan, Xf., Chen, F., Wu, Ss. et al. Proteomic analysis of differentially expressed proteins in mice with concanavalin A-induced hepatitis. J. Zhejiang Univ. Sci. B 11, 221–226 (2010). https://doi.org/10.1631/jzus.B0900351
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DOI: https://doi.org/10.1631/jzus.B0900351