• 1. West HJ. Clinical and pathological studies in horses with hepatic disease. Equine Vet J 1996; 28:146156.

  • 2. Durham AE, Newton JR, Smith KC, et al. Retrospective analysis of historical, clinical, ultrasonographic, serum biochemical and haematological data in prognostic evaluation of equine liver disease. Equine Vet J 2003; 35:542547.

    • Search Google Scholar
    • Export Citation
  • 3. Hackett ES, Twedt DC, Gustafson DL. Milk thistle and its derivative compounds: a review of opportunities for treatment of liver disease. J Vet Intern Med 2013; 27:1016.

    • Search Google Scholar
    • Export Citation
  • 4. Zhao J, Agarwal R. Tissue distribution of silibinin, the major active constituent of silymarin, in mice and its association with enhancement of phase II enzymes: implications in cancer chemoprevention. Carcinogenesis 1999; 20:21012108.

    • Search Google Scholar
    • Export Citation
  • 5. Huang D, Ou B, Prior RL. The chemistry behind antioxidant capacity assays. J Agric Food Chem 2005; 53:18411856.

  • 6. Kinnunen S, Hyyppa S, Lehmuskero A, et al. Oxygen radical absorbance capacity (ORAC) and exercise-induced oxidative stress in trotters. Eur J Appl Physiol 2005; 95:550556.

    • Search Google Scholar
    • Export Citation
  • 7. Strauss RG, Snyder EL, Wallace PD, et al. Oxygen-detoxifying enzymes in neutrophils of infants and their mothers. J Lab Clin Med 1980; 95:897904.

    • Search Google Scholar
    • Export Citation
  • 8. Vasiliou V, Ross D, Nebert DW. Update of the NAD(P)H:quinone oxidoreductase (NQO) gene family. Hum Genomics 2006; 2:329335.

  • 9. Alidoost F, Gharagozloo M, Bagherpour B, et al. Effects of silymarin on the proliferation and glutathione levels of peripheral blood mononuclear cells from beta-thalassemia major patients. Int Immunopharmacol 2006; 6:13051310.

    • Search Google Scholar
    • Export Citation
  • 10. Kiruthiga PV, Pandian SK, Devi KP. Silymarin protects PBMC against B(a)P induced toxicity by replenishing redox status and modulating glutathione metabolizing enzymes-an in vitro study. Toxicol Appl Pharmacol 2010; 247:116128.

    • Search Google Scholar
    • Export Citation
  • 11. Toklu HZ, Tunali Akbay T, Velioglu-Ogunc A, et al. Silymarin, the antioxidant component of Silybum marianum, prevents sepsis-induced acute lung and brain injury. J Surg Res 2008; 145:214222.

    • Search Google Scholar
    • Export Citation
  • 12. Polyak SJ, Morishima C, Shuhart MC, et al. Inhibition of T-cell inflammatory cytokines, hepatocyte NF-kappaB signaling, and HCV infection by standardized Silymarin. Gastroenterology 2007; 132:19251936.

    • Search Google Scholar
    • Export Citation
  • 13. Au AY, Hasenwinkel JM, Frondoza CG. Silybin inhibits interleukin-1beta-induced production of pro-inflammatory mediators in canine hepatocyte cultures. J Vet Pharmacol Ther 2011; 34:120129.

    • Search Google Scholar
    • Export Citation
  • 14. Tiegs G, Wolter M, Wendel A. Tumor necrosis factor is a terminal mediator in galactosamine/endotoxin-induced hepatitis in mice. Biochem Pharmacol 1989; 38:627631.

    • Search Google Scholar
    • Export Citation
  • 15. Kang YH, Berthiaume F, Yarmush ML. Long-term stable cultures of rat hepatocytes: an in vitro model to study acute and chronic hepatic inflammation. Tissue Eng 2002; 8:681693.

    • Search Google Scholar
    • Export Citation
  • 16. Fisher NC, Neil DA, Williams A, et al. Serum concentrations and peripheral secretion of the beta chemokines monocyte chemoattractant protein 1 and macrophage inflammatory protein 1alpha in alcoholic liver disease. Gut 1999; 45:416420.

    • Search Google Scholar
    • Export Citation
  • 17. Daniluk J, Szuster-Ciesielska A, Drabko J, et al. Serum cytokine levels in alcohol-related liver cirrhosis. Alcohol 2001; 23:2934.

    • Search Google Scholar
    • Export Citation
  • 18. Zi X, Mukhtar H, Agarwal R. Novel cancer chemopreventive effects of a flavonoid antioxidant silymarin: inhibition of mRNA expression of an endogenous tumor promoter TNF alpha. Biochem Biophys Res Commun 1997; 239:334339.

    • Search Google Scholar
    • Export Citation
  • 19. Schümann J, Wolf D, Pahl A, et al. Importance of Kupffer cells for T-cell-dependent liver injury in mice. Am J Pathol 2000; 157:16711683.

    • Search Google Scholar
    • Export Citation
  • 20. Hackett ES, Mama KR, Twedt DC, et al. Pharmacokinetics and safety of silibinin in horses. Am J Vet Res 2013; 74:13271332.

  • 21. Ungvari Z, Sosnowska D, Podlutsky A, et al. Free radical production, antioxidant capacity, and oxidative stress response signatures in fibroblasts from Lewis dwarf rats: effects of life span-extending peripubertal GH treatment. J Gerontol A Biol Sci Med Sci 2011; 66:501510.

    • Search Google Scholar
    • Export Citation
  • 22. Ou B, Hampsch-Woodill M, Prior RL. Development and validation of an improved oxygen radical absorbance capacity assay using fluorescein as the fluorescent probe. J Agric Food Chem 2001; 49:46194626.

    • Search Google Scholar
    • Export Citation
  • 23. Ninfali P, Aluigi G. Variability of oxygen radical absorbance capacity (ORAC) in different animal species. Free Radic Res 1998; 29:399408.

    • Search Google Scholar
    • Export Citation
  • 24. Gustafson DL, Siegel D, Rastatter JC, et al. Kinetics of NAD(P)H: quinone oxidoreductase I (NQO1) inhibition by mitomycin C in vitro and in vivo. J Pharmacol Exp Ther 2003; 305:10791086.

    • Search Google Scholar
    • Export Citation
  • 25. Gustafson DL, Swanson JD, Pritsos CA. Modulation of glutathione and glutathione dependent antioxidant enzymes in mouse heart following doxorubicin therapy. Free Radic Res Commun 1993; 19:111120.

    • Search Google Scholar
    • Export Citation
  • 26. Janiak M, Suska M, Dudzinska W, et al. Blood glutathione status and activity of glutathione-metabolizing antioxidant enzymes in erythrocytes of young trotters in basic training. J Anim Physiol Anim Nutr (Berl) 2010; 94:137145.

    • Search Google Scholar
    • Export Citation
  • 27. Adams AA, Katepalli MP, Kohler K, et al. Effect of body condition, body weight and adiposity on inflammatory cytokine responses in old horses. Vet Immunol Immunopathol 2009; 127:286294.

    • Search Google Scholar
    • Export Citation
  • 28. Breathnach CC, Sturgill-Wright T, Stiltner JL, et al. Foals are interferon gamma-deficient at birth. Vet Immunol Immunopathol 2006; 112:199209.

    • Search Google Scholar
    • Export Citation
  • 29. Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(−Delta Delta C(T)) Method. Methods 2001; 25:402408.

    • Search Google Scholar
    • Export Citation
  • 30. Kiruthiga PV, Shafreen RB, Pandian SK, et al. Silymarin protection against major reactive oxygen species released by environmental toxins: exogenous H2O2 exposure in erythrocytes. Basic Clin Pharmacol Toxicol 2007; 100:414419.

    • Search Google Scholar
    • Export Citation
  • 31. Roozbeh J, Shahriyari B, Akmali M, et al. Comparative effects of silymarin and vitamin E supplementation on oxidative stress markers, and hemoglobin levels among patients on hemodialysis. Ren Fail 2011; 33:118123.

    • Search Google Scholar
    • Export Citation
  • 32. Turgut F, Bayrak O, Catal F, et al. Antioxidant and protective effects of silymarin on ischemia and reperfusion injury in the kidney tissues of rats. Int Urol Nephrol 2008; 40:453460.

    • Search Google Scholar
    • Export Citation
  • 33. Webb CB, McCord KW, Twedt DC. Assessment of oxidative stress in leukocytes and granulocyte function following oral administration of a silibinin-phosphatidylcholine complex in cats. Am J Vet Res 2009; 70:5762.

    • Search Google Scholar
    • Export Citation
  • 34. Valenzuela A, Aspillaga M, Vial S, et al. Selectivity of silymarin on the increase of the glutathione content in different tissues of the rat. Planta Med 1989; 55:420422.

    • Search Google Scholar
    • Export Citation
  • 35. Das SK, Vasudevan DM. Protective effects of silymarin, a milk thistle (Silybium marianum) derivative on ethanolinduced oxidative stress in liver. Indian J Biochem Biophys 2006; 43:306311.

    • Search Google Scholar
    • Export Citation
  • 36. Pradeep K, Mohan CV, Gobianand K, et al. Silymarin modulates the oxidant-antioxidant imbalance during diethylnitrosamine induced oxidative stress in rats. Eur J Pharmacol 2007; 560:110116.

    • Search Google Scholar
    • Export Citation
  • 37. Campos R, Garrido A, Guerra R, et al. Silybin dihemisuccinate protects against glutathione depletion and lipid peroxidation induced by acetaminophen on rat liver. Planta Med 1989; 55:417419.

    • Search Google Scholar
    • Export Citation
  • 38. Yao J, Zhi M, Minhu C. Effect of silybin on high-fat-induced fatty liver in rats. Braz J Biol Med Res 2011; 44:652659.

  • 39. Fahmy SR, Hamdi SA. Curative effect of the Egyptian marine Erugosquilla massavensis extract on carbon tetrachloride-induced oxidative stress in rat liver and erythrocytes. Eur Rev Med Pharmacol Sci 2011; 15:303312.

    • Search Google Scholar
    • Export Citation
  • 40. Haddad Y, Vallerand D, Brault A, et al. Antioxidant and hepatoprotective effects of silibinin in a rat model of nonalcoholic steatohepatitis. Evid Based Complement Alternat Med 2011;2011: nep164.

    • Search Google Scholar
    • Export Citation
  • 41. Horváth ME, González-Cabello R, Blázovics A, et al. Effect of silibinin and vitamin E on restoration of cellular immune response after partial hepatectomy. J Ethnopharmacol 2001; 77:227232.

    • Search Google Scholar
    • Export Citation
  • 42. Al-Anati L, Essid E, Reinehr R, et al. Silibinin protects OTA-mediated TNF-alpha release from perfused rat livers and isolated rat Kupffer cells. Mol Nutr Food Res 2009; 53:460466.

    • Search Google Scholar
    • Export Citation
  • 43. Trappoliere M, Caligiuri A, Schmid M, et al. Silybin, a component of sylimarin, exerts anti-inflammatory and anti-fibrogenic effects on human hepatic stellate cells. J Hepatol 2009; 50:11021111.

    • Search Google Scholar
    • Export Citation
  • 44. Dietzmann J, Thiel U, Ansorge S, et al. Thiol-inducing and immunoregulatory effects of flavonoids in peripheral blood mononuclear cells from patients with end-stage diabetic nephropathy. Free Radic Biol Med 2002; 33:13471354.

    • Search Google Scholar
    • Export Citation
  • 45. Loguercio C, Federico A, Trappoliere M, et al. The effect of a silybin-vitamin E-phospholipid complex on nonalcoholic fatty liver disease: a pilot study. Dig Dis Sci 2007; 52:23872395.

    • Search Google Scholar
    • Export Citation
  • 46. Federico A, Trappoliere M, Tuccillo C, et al. A new silybin-vitamin E-phospholipid complex improves insulin resistance and liver damage in patients with non-alcoholic fatty liver disease: preliminary observations. Gut 2006; 55:901902.

    • Search Google Scholar
    • Export Citation
  • 47. Federico A, Niosi M, Vecchio Blanco CD, et al. Emerging drugs for non-alcoholic fatty liver disease. Expert Opin Emerg Drugs 2008; 13:145158.

    • Search Google Scholar
    • Export Citation

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Evaluation of antioxidant capacity and inflammatory cytokine gene expression in horses fed silibinin complexed with phospholipid

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  • 1 Department of Clinical Sciences, College of Veterinary Medicine and Biological Sciences, Colorado State University, Fort Collins, CO. 80523.
  • | 2 Department of Clinical Sciences, College of Veterinary Medicine and Biological Sciences, Colorado State University, Fort Collins, CO. 80523.
  • | 3 Department of Clinical Sciences, College of Veterinary Medicine and Biological Sciences, Colorado State University, Fort Collins, CO. 80523.
  • | 4 Department of Clinical Sciences, College of Veterinary Medicine and Biological Sciences, Colorado State University, Fort Collins, CO. 80523.

Abstract

Objective—To evaluate antioxidant capacity and inflammatory cytokine gene expression in horses fed silibinin complexed with phospholipid.

Animals—5 healthy horses.

Procedures—Horses consumed increasing orally administered doses of silibinin phospholipid during 4 nonconsecutive weeks (0 mg/kg, 6.5 mg/kg, 13 mg/kg, and 26 mg/kg of body weight, twice daily for 7 days each week). Dose-related changes in plasma antioxidant capacity, peripheral blood cell glutathione concentration and antioxidant enzyme activities, and blood cytokine gene expression were evaluated.

Results—Plasma antioxidant capacity increased throughout the study period with increasing dose. Red blood cell nicotinamide adenine dinucleotide phosphate:quinone oxidoreductase I activity decreased significantly with increasing doses of silibinin phospholipid. No significant differences were identified in glutathione peroxidase activity, reduced glutathione or oxidized glutathione concentrations, or expression of tumor necrosis factor α, interleukin-1, or interleukin-2.

Conclusions and Clinical Relevance—Minor alterations in antioxidant capacity of healthy horses that consumed silibinin phospholipid occurred and suggest that further study in horses with liver disease is indicated.

Abstract

Objective—To evaluate antioxidant capacity and inflammatory cytokine gene expression in horses fed silibinin complexed with phospholipid.

Animals—5 healthy horses.

Procedures—Horses consumed increasing orally administered doses of silibinin phospholipid during 4 nonconsecutive weeks (0 mg/kg, 6.5 mg/kg, 13 mg/kg, and 26 mg/kg of body weight, twice daily for 7 days each week). Dose-related changes in plasma antioxidant capacity, peripheral blood cell glutathione concentration and antioxidant enzyme activities, and blood cytokine gene expression were evaluated.

Results—Plasma antioxidant capacity increased throughout the study period with increasing dose. Red blood cell nicotinamide adenine dinucleotide phosphate:quinone oxidoreductase I activity decreased significantly with increasing doses of silibinin phospholipid. No significant differences were identified in glutathione peroxidase activity, reduced glutathione or oxidized glutathione concentrations, or expression of tumor necrosis factor α, interleukin-1, or interleukin-2.

Conclusions and Clinical Relevance—Minor alterations in antioxidant capacity of healthy horses that consumed silibinin phospholipid occurred and suggest that further study in horses with liver disease is indicated.

Contributor Notes

Address correspondence to Dr. Hackett (Eileen.Hackett@colostate.edu).