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Licensed Unlicensed Requires Authentication Published by De Gruyter February 6, 2018

Determination of selenium during pathogenesis of hepatic fibrosis employing hydride generation and inductively coupled plasma mass spectrometry

  • Joseph George ORCID logo EMAIL logo
From the journal Biological Chemistry


Serum and liver selenium levels were studied during the pathogenesis of N-nitrosodimethylamine (NDMA) induced hepatic fibrosis in rats. The degree of fibrosis was assessed with Masson’s trichrome staining and quantifying collagen content in the liver. Lipid peroxides were measured in blood and liver samples and total glutathione and glutathione peroxidase were assayed in the liver tissue to evaluate oxidative stress. Interleukin-6 (IL-6) and transforming growth factor-β1 (TGF-β1) were measured in the serum. Selenium levels were determined using inductively coupled plasma-mass spectrometry (ICP-MS) after acid digestion and hydride generation of selenium. Serial administrations of NDMA produced well-developed fibrosis and early cirrhosis in the liver with 4-fold increase of total collagen content and deposition of collagen fibers. Blood and hepatic lipid peroxides, serum IL-6 and TGF-β1 were significantly increased. There was significant reduction in hepatic glutathione and glutathione peroxidase levels. Serum and liver selenium were remarkably decreased on all the days studied. The results suggest that decreased selenium and glutathione peroxidase contribute to the impairment of cellular antioxidant defense, which in turn results in oxidative stress and trigger pathogenesis of hepatic fibrosis. The study further demonstrated that ICP-MS with hydride generation technique is a reliable and sensitive method for determination of selenium in biological samples.


The author is thankful to Dr. C. K. Mathews, Director, Chemical Group, Indira Gandhi Center for Atomic Research (IGCAR), Kalpakkam, Tamil Nadu, India for his permission to use the PerkinElmer SCIEX ELAN Inductively Coupled Plasma-Mass Spectrometer for the measurement of selenium. The technical assistance of Dr. Lisa George and Ms. S. Vijayalakshmi, Chemical Group, IGCAR, Kalpakkam is gratefully acknowledged. This work was supported by the Indian Council of Medical Research (Funder Id: 10.13039/501100001411), New Delhi, India, by grant no. 3/1/2/3/(9201540)/92-NCD-III to the author.

  1. Conflicts of interest statement: The author does not have any conflicts of interest to declare in connection with this article.


Alegre, F., Pelegrin, P., and Feldstein, A.E. (2017). Inflammasomes in liver fibrosis. Semin. Liver Dis. 37, 119–127.10.1055/s-0037-1601350Search in Google Scholar

Boosalis, M.G. (2008). The role of selenium in chronic disease. Nutr. Clin. Pract. 23, 152–160.10.1177/0884533608314532Search in Google Scholar

Brenneisen, P., Steinbrenner, H., and Sies, H. (2005). Selenium, oxidative stress, and health aspects. Mol. Asp. Med. 26, 256–267.10.1016/j.mam.2005.07.004Search in Google Scholar

Brigelius-Flohé, R. and Maiorino, M. (2013). Glutathione peroxidases. Biochim. Biophys. Acta 1830, 3289–3303.10.1016/j.bbagen.2012.11.020Search in Google Scholar

Buckley, W.T., Budac, J.J., Godfrey, D.V., and Koenig, K.M. (1992). Determination of selenium by inductively coupled plasma mass spectrometry utilizing a new hydride generation sample introduction system. Anal. Chem. 64, 724–729.10.1021/ac00031a006Search in Google Scholar

Burk, R.F., Early, D.S., Hill, K.E., Palmer, I.S., and Boeglin, M.E. (1998). Plasma selenium in patients with cirrhosis. Hepatology 27, 794–798.10.1002/hep.510270322Search in Google Scholar

Casaril, M., Stanzial, A.M., Gabrielli, G.B., Capra, F., Zenari, L., Galassini, S., Moschini, G., Liu, N.Q., and Corrocher, R. (1989). Serum selenium in liver cirrhosis: correlation with markers of fibrosis. Clin. Chim. Acta 182, 221–227.10.1016/0009-8981(89)90080-6Search in Google Scholar

Dworkin, B.M., Rosenthal, W.S., Stahl, R.E., and Panesar, N.K. (1988). Decreased hepatic selenium content in alcoholic cirrhosis. Dig. Dis. Sci. 33, 1213–1217.10.1007/BF01536668Search in Google Scholar

George, J. (2003). Ascorbic acid concentrations in dimethylnitrosamine-induced hepatic fibrosis in rats. Clin. Chim. Acta 335, 39–47.10.1016/S0009-8981(03)00285-7Search in Google Scholar

George, J. (2008). Elevated serum beta-glucuronidase reflects hepatic lysosomal fragility following toxic liver injury in rats. Biochem. Cell Biol. 86, 235–243.10.1139/O08-038Search in Google Scholar

George, J. and Chandrakasan, G. (1996). Molecular characteristics of dimethylnitrosamine induced fibrotic liver collagen. Biochim. Biophys. Acta 1292, 215–222.10.1016/0167-4838(95)00202-2Search in Google Scholar

George, J., Rao, K.R., Stern, R., and Chandrakasan, G. (2001). Dimethylnitrosamine-induced liver injury in rats: the early deposition of collagen. Toxicology 156, 129–138.10.1016/S0300-483X(00)00352-8Search in Google Scholar

George, J., Tsutsumi, M., and Tsuchishima, M. (2017). MMP-13 deletion decreases profibrogenic molecules and attenuates N-nitrosodimethylamine induced liver injury and fibrosis in mice. J. Cell. Mol. Med. 21, 3821–3835.10.1111/jcmm.13304Search in Google Scholar

Gheita, T.A. and Kenawy, S.A. (2014). Measurement of malondialdehyde, glutathione, and glutathione peroxidase in SLE patients. Methods Mol. Biol. 1134, 193–199.10.1007/978-1-4939-0326-9_14Search in Google Scholar

González-Reimers, E., Galindo-Martín, L., Santolaria-Fernández, F., Sánchez-Pérez, M.J., Alvisa-Negrín, J., García-Valdecasas-Campelo, E., González-Pérez, J.M., and Martín-González, M.C. (2008). Prognostic value of serum selenium levels in alcoholics. Biol. Trace Elem. Res. 125, 22–29.10.1007/s12011-008-8152-5Search in Google Scholar

Gutiérrez, R., Alvarado, J.L., Presno, M., Pérez-Veyna, O., Serrano, C.J., and Yahuaca, P. (2010). Oxidative stress modulation by Rosmarinus officinalis in CCl4-induced liver cirrhosis. Phytother. Res. 24, 595–601.10.1002/ptr.2997Search in Google Scholar

Jamall, I.S., Finelli, V.N., and Que Hee, S.S. (1981). A simple method to determine nanogram levels of 4-hydroxyproline in biological tissues. Anal. Biochem. 112, 70–75.10.1016/0003-2697(81)90261-XSearch in Google Scholar

Khan, N., Jeong, I.S., Hwang, I.M., Kim, J.S., Choi, S.H., Nho, E.Y., Choi, J.Y., Park, K.S., and Kim, K.S. (2014). Analysis of minor and trace elements in milk and yogurts by inductively coupled plasma-mass spectrometry (ICP-MS). Food Chem. 147, 220–224.10.1016/j.foodchem.2013.09.147Search in Google Scholar PubMed

Korpela, H., Kumpulainen, J., Luoma, P.V., Arranto, A.J., and Sotaniemi, E.A. (1985). Decreased serum selenium in alcoholics as related to liver structure and function. Am. J. Clin. Nutr. 42, 147–151.10.1093/ajcn/42.1.147Search in Google Scholar PubMed

Laclaustra, M., Navas-Acien, A., Stranges, S., Ordovas, J.M., and Guallar, E. (2009). Serum selenium concentrations and hypertension in the US Population. Circ. Cardiovasc. Qual. Outcomes 2, 369–376.10.1161/CIRCOUTCOMES.108.831552Search in Google Scholar PubMed PubMed Central

Lin, X., Chen, Y., Lv, S., Tan, S., Zhang, S., Huang, R., Zhuo, L., Liang, S., Lu, Z., and Huang, Q. (2015). Gypsophila elegans isoorientin attenuates CCl4-induced hepatic fibrosis in rats via modulation of NF-κB and TGF-β1/Smad signaling pathways. Int. Immunopharmacol. 28, 305–312.10.1016/j.intimp.2015.06.021Search in Google Scholar PubMed

Liu, R.M. and Gaston Pravia, K.A. (2010). Oxidative stress and glutathione in TGF-β-mediated fibrogenesis. Free Radic. Biol. Med. 48, 1–15.10.1016/j.freeradbiomed.2009.09.026Search in Google Scholar PubMed PubMed Central

Lu, C.W., Chang, H.H., Yang, K.C., Kuo, C.S., Lee, L.T., and Huang, K.C. (2016). High serum selenium levels are associated with increased risk for diabetes mellitus independent of central obesity and insulin resistance. BMJ Open Diabetes Res. Care 4, e000253.10.1136/bmjdrc-2016-000253Search in Google Scholar PubMed PubMed Central

Lubos, E., Loscalzo, J., and Handy, D.E. (2011). Glutathione peroxidase-1 in health and disease: from molecular mechanisms to therapeutic opportunities. Antioxid. Redox Signal. 15, 1957–1997.10.1089/ars.2010.3586Search in Google Scholar PubMed PubMed Central

Massadeh, A., Gharibeh, A., Omari, K., Al-Momani, I., Alomary, A., Tumah, H., and Hayajneh, W. (2010). Simultaneous determination of Cd, Pb, Cu, Zn, and Se in human blood of jordanian smokers by ICP-OES. Biol. Trace Elem. Res. 133, 1–11.10.1007/s12011-009-8405-ySearch in Google Scholar PubMed

Mormone, E., George, J., and Nieto, N. (2011). Molecular pathogenesis of hepatic fibrosis and current therapeutic approaches. Chem. Biol. Interact. 193, 225–231.10.1016/j.cbi.2011.07.001Search in Google Scholar PubMed PubMed Central

Nakahara, T. (1983). Applications of hydride generation techniques in atomic absorption, atomic fluorescence and plasma atomic emission spectroscopy. Prog. Anal. At. Spectrosc. 6, 163–223.Search in Google Scholar

Ojeda, M.L., Carreras, O., Sobrino, P., Murillo, M.L., and Nogales, F. (2017). Biological implications of selenium in adolescent rats exposed to binge drinking: oxidative, immunologic and apoptotic balance. Toxicol. Appl. Pharmacol. 329, 165–172.10.1016/j.taap.2017.05.037Search in Google Scholar PubMed

Pemberton, P.W., Smith, A., and Warnes, T.W. (2005). Non-invasive monitoring of oxidant stress in alcoholic liver disease. Scand. J. Gastroenterol. 40, 1102–1108.10.1080/00365520510023495Search in Google Scholar PubMed

Petrovski, B.E., Pataki, V., Jenei, T., Adány, R., and Vokó, Z. (2012). Selenium levels in men with liver disease in Hungary. J. Trace Elem. Med. Biol. 26, 31–35.10.1016/j.jtemb.2012.01.001Search in Google Scholar PubMed

Prystupa, A., Kiciński, P., Luchowska-Kocot, D., Błażewicz, A., Niedziałek, J., Mizerski, G., Jojczuk, M., Ochal, A., Sak, J.J., and Załuska, W. (2017). Association between serum selenium concentrations and levels of proinflammatory and profibrotic cytokines-interleukin-6 and growth differentiation factor-15, in patients with alcoholic liver cirrhosis. Int. J. Environ. Res. Public Health 14, E437.10.3390/ijerph14040437Search in Google Scholar PubMed PubMed Central

Ribas, V., García-Ruiz, C., and Fernández-Checa, J.C. (2014). Glutathione and mitochondria. Front. Pharmacol. 5, 151.10.3389/fphar.2014.00151Search in Google Scholar

Richter, K. and Kietzmann, T. (2016). Reactive oxygen species and fibrosis: further evidence of a significant liaison. Cell Tissue Res. 365, 591–605.10.1007/s00441-016-2445-3Search in Google Scholar

Rua, R.M., Ojeda, M.L., Nogales, F., Rubio, J.M., Romero-Gómez, M., Funuyet, J., Murillo, M.L., and Carreras, O. (2014). Serum selenium levels and oxidative balance as differential markers in hepatic damage caused by alcohol. Life Sci. 94, 158–163.10.1016/j.lfs.2013.10.008Search in Google Scholar

Sánchez-Valle, V., Chávez-Tapia, N.C., Uribe, M., and Méndez-Sánchez, N. (2012). Role of oxidative stress and molecular changes in liver fibrosis: a review. Curr. Med. Chem. 19, 4850–4860.10.2174/092986712803341520Search in Google Scholar

Schmidt-Arras, D. and Rose-John, S. (2016). IL-6 pathway in the liver: from physiopathology to therapy. J. Hepatol. 64, 1403–1415.10.1016/j.jhep.2016.02.004Search in Google Scholar

Seki, E. and Brenner, D.A. (2015). Recent advancement of molecular mechanisms of liver fibrosis. J. Hepatobiliary Pancreat. Sci. 22, 512–518.10.1002/jhbp.245Search in Google Scholar

Thuluvath, P.J. and Triger, D.R. (1992). Selenium in chronic liver disease. J. Hepatol. 14, 176–182.10.1016/0168-8278(92)90155-ISearch in Google Scholar

Tinggi, U. (2008). Selenium: its role as antioxidant in human health. Environ. Health Prev. Med. 13, 102–108.10.1007/s12199-007-0019-4Search in Google Scholar

Valimaki, M.J., Harju, K.J., and Ylikahri, R.H. (1983). Decreased serum selenium in alcoholics – a consequence of liver dysfunction. Clin. Chim. Acta 130, 291–296.10.1016/0009-8981(83)90303-0Search in Google Scholar

Vijayalakshmi, S., Prabhu, R.K., Mahalingam, T.R., and Mathews, C.K. (1992). A simple gas-liquid separator for continuous hydride introduction in ICP-MS. At. Spectrosc. 13, 26–28.Search in Google Scholar

Watkinson, J.H. (1966). Fluorometric determination of selenium in biological material with 2,3-diaminonaphthalene. Anal. Chem. 38, 92–97.10.1021/ac60233a025Search in Google Scholar PubMed

Zhang, C.Y., Yuan, W.G., He, P., Lei, J.H., and Wang, C.X. (2016). Liver fibrosis and hepatic stellate cells: etiology, pathological hallmarks and therapeutic targets. World J. Gastroenterol. 22, 10512–10522.10.3748/wjg.v22.i48.10512Search in Google Scholar PubMed PubMed Central

Received: 2017-10-04
Accepted: 2018-01-11
Published Online: 2018-02-06
Published in Print: 2018-04-25

©2018 Walter de Gruyter GmbH, Berlin/Boston

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