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Planta Med 2013; 79(12): 997-1002
DOI: 10.1055/s-0032-1328704
Biological and Pharmacological Activity
Original Papers
Georg Thieme Verlag KG Stuttgart · New York

Alterations in Sulfur Amino Acid Metabolism in Mice Treated with Silymarin: A Novel Mechanism of Its Action Involved in Enhancement of the Antioxidant Defense in Liver

Do Young Kwon
1   College of Pharmacy, Seoul National University, Seoul, Korea
,
Young Suk Jung
1   College of Pharmacy, Seoul National University, Seoul, Korea
,
Sun Ju Kim
1   College of Pharmacy, Seoul National University, Seoul, Korea
,
Young Soon Kim
2   Department of Food & Nutrition, College of Health Sciences, Korea University, Seoul, Korea
,
Dal Woong Choi
3   Department of Environmental Health, College of Health Sciences, Korea University, Seoul, Korea
,
Young Chul Kim
1   College of Pharmacy, Seoul National University, Seoul, Korea
4   Research Institute of Pharmaceutical Sciences, Seoul National University, Seoul, Korea
› Author Affiliations
Further Information

Publication History

received 14 March 2013
revised 20 May 2013

accepted 24 May 2013

Publication Date:
27 June 2013 (online)

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Abstract

It has been known that silymarin exhibits protective activity against oxidative liver injury induced by various hepatotoxicants, but the underlying mechanism of its beneficial action remains unclear. We determined the alterations in sulfur-containing amino acid metabolism induced by silymarin in association with its effects on the antioxidant capacity of liver. Male mice were treated with silymarin (100 or 200 mg/kg, p. o.) every 12 h for a total of 3 doses, and sacrificed 6 h after the final dosing. The hepatic methionine level was increased, but the activity and protein expression of methionine adenosyltransferase were decreased by silymarin in a dose-dependent manner. S-Adenosylmethionine or homocysteine concentration was not changed, whereas the sulfur-containing metabolites generated from homocysteine in the transsulfuration pathway including cystathionine, cysteine, and glutathione were increased significantly. Cystathionine β-synthase was induced, but cysteine dioxygenase was downregulated, both of which would contribute to the elevation of cysteine and its product, glutathione, in liver. Oxygen radical scavenging capacity of liver cytosol against peroxyl radical and peroxynitrite was increased, and also hepatic lipid peroxidation was diminished in the silymarin-treated mice. Taken together, the results demonstrate that silymarin enhances hepatic glutathione generation by elevating cysteine availability via an increment in cysteine synthesis and an inhibition of its catabolism to taurine, which may subsequently contribute to the antioxidant defense of liver.

Key words

Silybum marianum - Asteraceae - silymarin - glutathione - sulfur amino acid metabolism - antioxidant
 

  • References

  • 1 Song Z, Deaciuc I, Song M, Lee DY, Liu Y, Ji X, McClain C. Silymarin protects against acute ethanol-induced hepatotoxicity in mice. Alcohol Clin Exp Res 2006; 30: 407-413
    CrossrefPubMedGoogle Scholar
  • 2 Pradeep K, Mohan CV, Gobianand K, Karthikeyan S. Silymarin modulates the oxidant-antioxidant imbalance during diethylnitrosamine induced oxidative stress in rats. Eur J Pharmacol 2007; 560: 110-116
    CrossrefPubMedGoogle Scholar
  • 3 Lettéron P, Labbe G, Degott C, Berson A, Fromenty B, Delaforge M, Larrey D, Pessayre D. Mechanism for the protective effects of silymarin against carbon tetrachloride-induced lipid peroxidation and hepatotoxicity in mice. Evidence that silymarin acts both as an inhibitor of metabolic activation and as a chain-breaking antioxidant. Biochem Pharmacol 1990; 39: 2027-2034
    CrossrefPubMedGoogle Scholar
  • 4 Bindoli A, Cavallini L, Siliprandi N. Inhibitory action of silymarin of lipid peroxide formation in rat liver mitochondria and microsomes. Biochem Pharmacol 1977; 26: 2405-2409
    CrossrefPubMedGoogle Scholar
  • 5 Varga Z, Seres I, Nagy E, Ujhelyi L, Balla G, Balla J, Antus S. Structure prerequisite for antioxidant activity of silybin in different biochemical systems in vitro . Phytomedicine 2006; 13: 85-93
    CrossrefPubMedGoogle Scholar
  • 6 Dehmlow C, Murawski N, de Groot H. Scavenging of reactive oxygen species and inhibition of arachidonic acid metabolism by silibinin in human cells. Life Sci 1996; 58: 1591-1600
    CrossrefPubMedGoogle Scholar
  • 7 Comoglio A, Tomasi A, Malandrino S, Poli G, Albano E. Scavenging effect of silipide, a new silybin-phospholipid complex, on ethanol-derived free radicals. Biochem Pharmacol 1995; 50: 1313-1316
    CrossrefPubMedGoogle Scholar
  • 8 Valenzuela A, Lagos C, Schmidt K, Videla LA. Silymarin protection against hepatic lipid peroxidation induced by acute ethanol intoxication in the rat. Biochem Pharmacol 1985; 34: 2209-2212
    CrossrefPubMedGoogle Scholar
  • 9 Valenzuela A, Aspillaga M, Vial S, Guerra R. Selectivity of silymarin on the increase of the glutathione content in different tissues of the rat. Planta Med 1989; 55: 420-422
    Article in Thieme ConnectPubMedGoogle Scholar
  • 10 Kim SJ, Lee JW, Jung YS, Kwon DY, Park HK, Ryu CS, Kim SK, Oh GT, Kim YC. Ethanol-induced liver injury and changes in sulfur amino acid metabolomics in glutathione peroxidase and catalase double knockout mice. J Hepatol 2009; 50: 1184-1191
    CrossrefPubMedGoogle Scholar
  • 11 Ratnam S, Maclean KN, Jacobs RL, Brosnan ME, Kraus JP, Brosnan JT. Hormonal regulation of cystathionine beta-synthase expression in liver. J Biol Chem 2002; 277: 42912-42918
    CrossrefPubMedGoogle Scholar
  • 12 Pradhan SC, Girish C. Hepatoprotective herbal drug, silymarin from experimental pharmacology to clinical medicine. Indian J Med Res 2006; 124: 491-504
    PubMedGoogle Scholar
  • 13 Stipanuk MH, Hirschberger LL, Londono MP, Cresenzi CL, Yu AF. The ubiquitin-proteasome system is responsible for cysteine-responsive regulation of cysteine dioxygenase concentration in liver. Am J Physiol Endocrinol Metab 2004; 286: E439-E448
    PubMedGoogle Scholar
  • 14 Kwon YH, Stipanuk MH. Cysteine regulates expression of cysteine dioxygenase and gamma-glutamylcysteine synthetase in cultured rat hepatocytes. Am J Physiol Endocrinol Metab 2001; 280: E804-E815
    PubMedGoogle Scholar
  • 15 Ueki I, Stipanuk MH. 3 T3-L1 adipocytes and rat adipose tissue have a high capacity for taurine synthesis by the cysteine dioxygenase/cysteinesulfinate decarboxylase and cysteamine dioxygenase pathways. J Nutr 2009; 139: 207-214
    PubMedGoogle Scholar
  • 16 Ueki I, Roman HB, Hirschberger LL, Junior C, Stipanuk MH. Extrahepatic tissues compensate for loss of hepatic taurine synthesis in mice with liver-specific knockout of cysteine dioxygenase. Am J Physiol Endocrinol Metab 2012; 302: E1292-E1299
    CrossrefPubMedGoogle Scholar
  • 17 Hosokawa Y, Yamaguchi K, Kohashi N, Kori Y, Ueda I. Decrease of rat liver cysteine dioxygenase (cysteine oxidase) activity mediated by glucagon. J Biochem 1978; 84: 419-424
    PubMedGoogle Scholar
  • 18 Crocenzi FA, Basiglio CL, Pérez LM, Portesio MS, Pozzi EJ, Roma MG. Silibinin prevents cholestasis-associated retrieval of the bile salt export pump, Bsep, in isolated rat hepatocyte couplets: possible involvement of cAMP. Biochem Pharmacol 2005; 69: 1113-1120
    CrossrefPubMedGoogle Scholar
  • 19 Scharf G, Prustomersky S, Knasmuller S, Schulte-Hermann R, Huber WW. Enhancement of glutathione and γ-glutamylcysteine synthetase, the rate limiting enzyme of glutathione synthesis, by chemoprotective plant-derived food and beverage components in the human hepatoma cell line HepG2. Nutr Cancer 2003; 45: 74-83
    CrossrefPubMedGoogle Scholar
  • 20 Myhrstad MC, Carlsen H, Nordström O, Blomhoff R, Moskaug JØ. Flavonoids increase the intracellular glutathione level by transactivation of the gamma-glutamylcysteine synthetase catalytical subunit promoter. Free Radic Biol Med 2002; 32: 386-393
    CrossrefPubMedGoogle Scholar
  • 21 Griffith OW. Determination of glutathione and glutathione disulfide using glutathione reductase and 2-vinylpyridine. Anal Biochem 1980; 106: 207-212
    CrossrefPubMedGoogle Scholar
  • 22 Gaitonde MK. A spectrophotometric method for the direct determination of cysteine in the presence of other naturally occurring amino acids. Biochem J 1976; 104: 627-633
    PubMedGoogle Scholar
  • 23 Rajendra W. High performance liquid chromatographic determination of amino acids in biological samples by precolumn derivatization with O-phthaldehyde. J Liq Chromatogr 1987; 10: 941-955
    PubMedGoogle Scholar
  • 24 She QB, Nagao I, Hayakawa T, Tsuge H. A simple HPLC method for the determination of S-adenosylmethionine and S-adenosylhomocysteine in rat tissues: the effect of vitamin B6 deficiency on these concentrations in rat liver. Biochem Biophys Res Commun 1994; 205: 1748-1754
    CrossrefPubMedGoogle Scholar
  • 25 Nolin TD, McMenamin ME, Himmelfarb J. Simultaneous determination of total homocysteine, cysteine, cysteinylglycine, and glutathione in human plasma by high-performance liquid chromatography: application to studies of oxidative stress. J Chromatogr B Analyt Technol Biomed Life Sci 2007; 852: 554-561
    PubMedGoogle Scholar
  • 26 Ericson LE, Harper AE. Effect of diet on the betaine-homocysteine transmethylase activity of rat liver. I. Amino acids and protein. J Biol Chem 1956; 219: 49-58
    PubMedGoogle Scholar
  • 27 Kashiwamata S, Greenberg DM. Studies on cystathionine synthase of rat liver. Properties of the highly purified enzyme. Biochim Biophys Acta 1970; 212: 488-500
    CrossrefPubMedGoogle Scholar
  • 28 Matsuo Y, Greenberg DM. A crystalline enzyme that cleaves homoserine and cystathionine. I. Isolation procedure and some physicochemical properties. J Biol Chem 1958; 230: 545-560
    PubMedGoogle Scholar
  • 29 Germanò MP, De Pasquale R, DʼAngelo V, Catania S, Silvari V, Costa C. Evaluation of extracts and isolated fraction from Capparis spinosa L. buds as an antioxidant source. J Agric Food Chem 2002; 50: 1168-1171
    CrossrefPubMedGoogle Scholar
  • 30 Regoli F, Winston GW. Quantification of total oxidant scavenging capacity of antioxidants for peroxynitrite, peroxyl radicals, and hydroxyl radicals. Toxicol Appl Pharmacol 1999; 156: 96-105
    CrossrefPubMedGoogle Scholar