Discovery of serum biomarkers of alcoholic fatty liver in a rodent model: C-reactive protein

Journal of Biomedical Science

Jungshan Chang⁷  Ling-Yun Chen⁵  Ai-Sheng Ho⁶  Shui-Cheng Lee¹  Chia-Chi Wang³  Fu-Der Mai⁴  Chun-Chao Chang²  Chun-Chia Cheng¹  Shu-Lin Liu⁵ 

[1] Institute of Nuclear Energy Research, Atomic Energy Council, Taoyuan, Taiwan;Department of Internal Medicine, School of Medicine, College of Medicine, Taipei Medical University Hospital, Taipei, Taiwan;Division of Gastroenterology, Buddhist Tzu Chi General Hospital, Taipei branch, Taiwan;Biomedical Mass Imaging Research Center, Taipei Medical University, Taipei, Taiwan;Institute of Biochemistry and Biotechnology, Chung Shan Medical University, Taichung, Taiwan;Division of Gastroenterology, Cheng Hsin General Hospital, Taipei, Taiwan;Research Center For Biomedical Implants and Microsurgery Devices, Taipei Medical University, Taipei, Taiwan

关键词: two-dimensional differential gel electrophoresis;    haptoglobin;    C - reactive protein;    biomarker;    alcoholic fatty liver;   

Others  :  833154
DOI  :  10.1186/1423-0127-18-52

 received in 2011-03-25, accepted in 2011-08-01,  发布年份 2011

【 摘 要 】

Background

Excessive consumption of alcohol contributes to alcoholic liver disease. Fatty liver is the early stage of alcohol-related liver disease. The aim of this study was to search for specific serological biomarkers of alcoholic fatty liver (AFL) compared to healthy controls, non-alcoholic fatty liver (NAFL) and liver fibrosis in a rodent model.

Methods

Serum samples derived from animals with AFL, NAFL, or liver fibrosis were characterized and compared using two-dimensional differential gel electrophoresis. A matrix-assisted laser desorption ionization-time of flight tandem mass spectrometer in conjunction with mascot software was used for protein identification. Subsequently, Western blotting and flexible multi-analyte profiling were used to measure the expressions of the putative biomarkers present in the serum of animals and clinical patients.

Results

Eight differential putative biomarkers were identified, and the two most differentiated proteins, including upregulated C-reactive protein (CRP) and downregulated haptoglobin (Hp), were further investigated. Western blotting validated that CRP was dramatically higher in the serum of AFL compared to healthy controls and other animals with liver disease of NAFL or liver fibrosis (p < 0.05). Moreover, we found that CRP and Hp were both lower in liver fibrosis of TAA-induced rats and clinical hepatitis C virus-infected patients.

Conclusion

The results suggest that increased levels of CRP are an early sign of AFL in rats. The abnormally elevated CRP induced by ethanol can be used as a biomarker to distinguish AFL from normal or otherwise diseased livers.

【 授权许可】

   
2011 Liu et al; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

Figure 2.

Figure 3.

Figure 4.

【 参考文献 】

• [1]Rouach H, Fataccioli V, Gentil M, French SW, Morimoto M, Nordmann R: Effect of chronic ethanol feeding on lipid peroxidation and protein oxidation in relation to liver pathology. Hepatology 1997, 25:351-355.
• [2]Grunnet N, Kondrup J, Dich J: Effect of ethanol on lipid metabolism in cultured hepatocytes. Biochem J 1985, 228:673-681.
• [3]Maharaj B, Maharaj RJ, Leary WP, Cooppan RM, Naran AD, Pirie D, Pudifin DJ: Sampling variability and its influence on the diagnostic yield of percutaneous needle biopsy of the liver. Lancet 1986, 1:523-525.
• [4]Dey A, Cederbaum AI: Alcohol and oxidative liver injury. Hepatology 2006, 43:S63-74.
• [5]Levitsky J, Mailliard ME: Diagnosis and therapy of alcoholic liver disease. Semin Liver Dis 2004, 24:233-247.
• [6]Fujimoto M, Uemura M, Kojima H, Ishii Y, Ann T, Sakurai S, Okuda K, Noguchi R, Adachi S, Kitano H, et al.: Prognostic factors in severe alcoholic liver injury. Nara Liver Study Group. Alcohol Clin Exp Res 1999, 23:33S-38S.
• [7]Vanbiervliet G, Le Breton F, Rosenthal-Allieri MA, Gelsi E, Marine-Barjoan E, Anty R, Piche T, Benzaken S, Saint-Paul MC, Huet PM, Tran A: Serum C-reactive protein: a non-invasive marker of alcoholic hepatitis. Scand J Gastroenterol 2006, 41:1473-1479.
• [8]Gupta S, Slaughter S, Akriviadis EA, Valenzuela R, Deodhar SD: Serial measurement of serum C-reactive protein facilitates evaluation in alcoholic hepatitis. Hepatogastroenterology 1995, 42:516-521.
• [9]Gonzalez-Quintela A, Mella C, Perez LF, Abdulkader I, Caparrini AM, Lojo S: Increased serum tissue polypeptide specific antigen (TPS) in alcoholics: a possible marker of alcoholic hepatitis. Alcohol Clin Exp Res 2000, 24:1222-1226.
• [10]Hill DB, Marsano L, Cohen D, Allen J, Shedlofsky S, McClain CJ: Increased plasma interleukin-6 concentrations in alcoholic hepatitis. J Lab Clin Med 1992, 119:547-552.
• [11]Unlu M, Morgan ME, Minden JS: Difference gel electrophoresis: a single gel method for detecting changes in protein extracts. Electrophoresis 1997, 18:2071-2077.
• [12]Byrne JC, Downes MR, O'Donoghue N, O'Keane C, O'Neill A, Fan Y, Fitzpatrick JM, Dunn M, Watson RW: 2D-DIGE as a strategy to identify serum markers for the progression of prostate cancer. J Proteome Res 2009, 8:942-957.
• [13]Ho AS, Cheng CC, Lee SC, Liu ML, Lee JY, Wang WM, Wang CC: Novel biomarkers predict liver fibrosis in hepatitis C patients: alpha 2 macroglobulin, vitamin D binding protein and apolipoprotein AI. J Biomed Sci 2010, 17:58. BioMed Central Full Text
• [14]Kondo T, Hirohashi S: Application of 2D-DIGE in cancer proteomics toward personalized medicine. Methods Mol Biol 2009, 577:135-154.
• [15]Orenes-Pinero E, Corton M, Gonzalez-Peramato P, Algaba F, Casal I, Serrano A, Sanchez-Carbayo M: Searching urinary tumor markers for bladder cancer using a two-dimensional differential gel electrophoresis (2D-DIGE) approach. J Proteome Res 2007, 6:4440-4448.
• [16]Keshavarzian A, Farhadi A, Forsyth CB, Rangan J, Jakate S, Shaikh M, Banan A, Fields JZ: Evidence that chronic alcohol exposure promotes intestinal oxidative stress, intestinal hyperpermeability and endotoxemia prior to development of alcoholic steatohepatitis in rats. J Hepatol 2009, 50:538-547.
• [17]Gentile M, Panico S, Rubba F, Mattiello A, Chiodini P, Jossa F, Marotta G, Pauciullo P, Rubba P: Obesity, overweight, and weight gain over adult life are main determinants of elevated hs-CRP in a cohort of Mediterranean women. Eur J Clin Nutr 2010, 64:873-878.
• [18]Oda E: CRP may be superior to anthropometric markers of obesity. Circ J 2007, 71:1332. author reply 1332-1333
• [19]Martinez VB, Gonzalez-Juanatey JR: Markers of inflammation and cardiovascular disease: clinical applications of C-reactive protein determination. Am J Cardiovasc Drugs 2009, 9(Suppl 1):3-7.
• [20]Ridker PM: Clinical application of C-reactive protein for cardiovascular disease detection and prevention. Circulation 2003, 107:363-369.
• [21]Albert MA, Glynn RJ, Ridker PM: Alcohol consumption and plasma concentration of C-reactive protein. Circulation 2003, 107:443-447.
• [22]Pepys MB, Hirschfield GM: C-reactive protein: a critical update. J Clin Invest 2003, 111:1805-1812.
• [23]Das T, Sen AK, Kempf T, Pramanik SR, Mandal C, Mandal C: Induction of glycosylation in human C-reactive protein under different pathological conditions. Biochem J 2003, 373:345-355.
• [24]Laskowska-Klita T, Czerwinska B: [Concentration of C-reactive protein, procalcitonin and alpha-1-antitrypsin in blood of neonates and infants with signs of inflammation]. Med Wieku Rozwoj 2002, 6:5-11.
• [25]Borawski J, Naumnik B, Mysliwiec M: Serum alpha1-antitrypsin but not complement C3 and C4 predicts chronic inflammation in hemodialysis patients. Ren Fail 2003, 25:589-593.
• [26]Cederbaum AI, Lu Y, Wu D: Role of oxidative stress in alcohol-induced liver injury. Arch Toxicol 2009, 83:519-548.
• [27]Nanji AA, Zhao S, Lamb RG, Dannenberg AJ, Sadrzadeh SM, Waxman DJ: Changes in cytochromes P-450, 2E1, 2B1, and 4A, and phospholipases A and C in the intragastric feeding rat model for alcoholic liver disease: relationship to dietary fats and pathologic liver injury. Alcohol Clin Exp Res 1994, 18:902-908.
• [28]Cao Q, Mak KM, Lieber CS: Cytochrome P4502E1 primes macrophages to increase TNF-alpha production in response to lipopolysaccharide. Am J Physiol Gastrointest Liver Physiol 2005, 289:G95-107.
• [29]Lu Y, Cederbaum AI: CYP2E1 and oxidative liver injury by alcohol. Free Radic Biol Med 2008, 44:723-738.
• [30]Naveau S, Cassard-Doulcier AM, Njike-Nakseu M, Bouchet-Delbos L, Barri-Ova N, Boujedidi H, Dauvois B, Balian A, Maitre S, Prevot S, et al.: Harmful effect of adipose tissue on liver lesions in patients with alcoholic liver disease. J Hepatol 2010, 52:895-902.
• [31]Chiellini C, Santini F, Marsili A, Berti P, Bertacca A, Pelosini C, Scartabelli G, Pardini E, Lopez-Soriano J, Centoni R, et al.: Serum haptoglobin: a novel marker of adiposity in humans. J Clin Endocrinol Metab 2004, 89:2678-2683.
• [32]Shu H, Kang X, Guo K, Li S, Li M, Sun L, Gan L, Liu Y, Qin X: Diagnostic value of serum haptoglobin protein as hepatocellular carcinoma candidate marker complementary to alpha fetoprotein. Oncol Rep 2010, 24:1271-1276.
• [33]Lee HH, Seo YS, Um SH, Won NH, Yoo H, Jung ES, Kwon YD, Park S, Keum B, Kim YS, et al.: Usefulness of non-invasive markers for predicting significant fibrosis in patients with chronic liver disease. J Korean Med Sci 2010, 25:67-74.
• [34]Thabut D, Naveau S, Charlotte F, Massard J, Ratziu V, Imbert-Bismut F, Cazals-Hatem D, Abella A, Messous D, Beuzen F, et al.: The diagnostic value of biomarkers (AshTest) for the prediction of alcoholic steato-hepatitis in patients with chronic alcoholic liver disease. J Hepatol 2006, 44:1175-1185.
• [35]Pasceri V, Willerson JT, Yeh ET: Direct proinflammatory effect of C-reactive protein on human endothelial cells. Circulation 2000, 102:2165-2168.