Original Article
  

Inhibition of Apoptosis and Proliferation in T Cells by Immunosuppressive Silymarine

Abstract

Silybum marianum, is known to have anti-inflammatory, hepatoprotective and anticarciogenic effects. The aim of this study was to compare effects of Silymarin, Rapamycin and FK506 on proliferation and apoptosis of human T cells stimulated with Con A. Peripheral blood mononuclear cells (PBMC) were stimulated with concavalin A (Con A) (5µg/mL) and then treated with different inhibitors (silymarin, rapamycin and FK506) in various concentrations (5 days). Cells were examined using carboxyfluorescein succinimidyl ester (CFSE) assay for proliferation. Then cell apoptosis was analyzed by FITC annexin V/PI staining and flow cytometry. The effects of drugs on the activation of poly ADP ribose polymerase (PARP) pathway in PBMCs stimulated with Con A and treated with IC50 dose of drugs for 5 days were evaluated using the PathScan cleaved PARP sandwich ELISA kit. The results indicated that silymarin inhibited T cell proliferation. In addition, our results pointed out that 100 μM and 200 μM of silymarin significantly have more inhibitory effect on T cells proliferation than FK506 and rapamycin. None of these drugs at IC50 concentration had affected the level of cleaved PARP. Overall, with superior efficacy and lesser toxicity in comparison with other immunosuppressive drugs, silymarin could be a suitable choice of therapy for certain diseases.

  1. Wattenberg L. Inhibition of Carcinogenesis by Naturally-Occurring and Synthetic Compounds. In: Kuroda Y, Shankel D, Waters M, editors. Antimutagenesis and Anticarcinogenesis Mechanisms II. Basic Life Sciences. 52: Springer US; 1990. p. 155-66.
  2. Valenzuela A, Garrido A. Biochemical bases of the pharmacological action of the flavonoid silymarin and of its structural isomer silibinin. Biol Res. 1994;27(2):105-12.
  3. Ahmed B, Ahmed B, Khan SA, Alam T. Synthesis and antihepatotoxic activity of some heterocyclic compounds containing the 1,4-dioxane ring system. Die Pharmazie - An International Journal of Pharmaceutical Sciences. 2003;58(3):173-6.
  4. Ball KR, Kowdley KV. A Review of Silybum marianum (Milk Thistle) as a Treatment for Alcoholic Liver Disease. Journal of Clinical Gastroenterology. 2005;39(6):520-8.
  5. Gazak R, Walterova D, Kren V. Silybin and silymarin--new and emerging applications in medicine. Curr Med Chem. 2007;14(3):315-38.
  6. Saller R, Meier R, Brignoli R. The Use of Silymarin in the Treatment of Liver Diseases. Drugs. 2001;61(14):2035-63.
  7. Polyak SJ, Morishima C, Shuhart MC, Wang CC, Liu Y, Lee DYW. Inhibition of T-Cell Inflammatory Cytokines, Hepatocyte NF-κB Signaling, and HCV Infection by Standardized Silymarin. Gastroenterology. 2007;132(5):1925.
  8. Alidoost F, Gharagozloo M, Bagherpour B, Jafarian A, Sajjadi SE, Hourfar H, et al. Effects of silymarin on the proliferation and glutathione levels of peripheral blood mononuclear cells from β-thalassemia major patients. International Immunopharmacology. 2006;6(8):1305-10.
  9. Gharagozloo M, Javid EN, Rezaei A, Mousavizadeh K. Silymarin inhibits cell cycle progression and mTOR activity in activated human T cells: therapeutic implications for autoimmune diseases. Basic & clinical pharmacology & toxicology. 2013;112(4):251-6.
  10. Wilasrusmee C, Kittur S, Shah G, Siddiqui J, Bruch D, Wilasrus- mee S et al. Immunostimulatory effect of Silybum marianum (milk thistle) extract. Med Sci Monit 2002;8:BR439–43.
  11. Gharagozloo M, Velardi E, Bruscoli S, Agostini M, Di Sante M, Donato V et al. Silymarin suppress CD4+ T cell activation and proliferation: effects on NF-kappaB activity and IL-2 production. Pharmacol Res 2010;61:405–9. 10
  12. Gharagozloo M, Javid EN, Rezaei A, Mousavizadeh K. Silymarin inhibits cell cycle progression and mTOR activity in activated human T cells: therapeutic implications for autoimmune diseases. Basic Clin Pharmacol Toxicol 2012;12:251–6. 11
  13. Morishima C, Shuhart MC, Wang CC, Paschal DM, Apodaca MC, Liu Y et al. Silymarin inhibits in vitro T-cell proliferation and cytokine production in hepatitis C virus infection. Gastroenterology 2010;138:671–81.
  14. Frohn C, Fricke L, Puchta JC, Kirchner H. The effect of HLA‐C matching on acute renal transplant rejection. Nephrology Dialysis Transplantation. 2001;16(2):355-60.
  15. Gharagozloo M, Velardi E, Bruscoli S, Agostini M, Di Sante M, Donato V, et al. Silymarin suppress CD4+ T cell activation and proliferation: Effects on NF-κB activity and IL-2 production. Pharmacological Research. 2010;61(5):405-9.
  16. Schümann J, Prockl J, Kiemer AK, Vollmar AM, Bang R, Tiegs G. Silibinin protects mice from T cell-dependent liver injury. Journal of Hepatology. 2003;39(3):333-40.
  17. Meroni PL, Barcellini W, Borghi MO, Vismara A, Ferraro G, Ciani D, et al. Silybin inhibition of human T-lymphocyte activation. Int J Tissue React. 1988;10(3):177-81.
  18. Morishima C, Shuhart MC, Wang CC, Paschal DM, Apodaca MC, Liu Y, et al. Silymarin Inhibits In Vitro T-Cell Proliferation and Cytokine Production in Hepatitis C Virus Infection. Gastroenterology. 2010;138(2):671-81.e2.
  19. Cruz T, Galvez J, Crespo E, Ocete MA, Zarzuelo A. Effects of silymarin on the acute stage of the trinitrobenzenesulphonic acid model of rat colitis. PlantaMed 2001;67(1):94–6.
  20. Johnson VJ, He Q, Osuchowski MF, Sharma RP. Physiological responses of a natural antioxidant flavonoid mixture, silymarin, in BALB/c mice: III. Silymarin inhibits T-lymphocyte function at low doses but stimulates inflammatory processes at high doses. Planta Med 2003;69(1):44–9.
  21. Gharagozloo M, Javid EN, Rezaei A, Mousavizadeh K. Silymarin inhibits cell cycle progression and mTOR activity in activated human T cells: therapeutic implications for autoimmune diseases. Basic & clinical pharmacology & toxicology. 2013;112(4):251-6.
  22. Gharagozloo M, Jafari S, Esmaeil N, Javid EN, Bagherpour B, Rezaei A. Immunosuppressive effect of silymarin on mitogen-activated protein kinase signalling pathway: the impact on T cell proliferation and cytokine production. Basic & clinical pharmacology & toxicology. 2013;113(3):209-14.
  23. Thornberry N and Lazebnik Y: Caspases: enemies within. Science 281: 1312-1316, 1998.
  24. Degen WG, Pruijn GJ, Raats JM and van Venrooij WJ: Caspase-dependent cleavage of nucleic acids. Cell Death Differ 7: 616-627, 2000
  25. Slee EA, Adrian C and Martin SJ: Serial killers: ordering caspase activation events in apoptosis. Cell Death Differ 6: 1067-1074, 1999
  26. Huppertz B, Frank H-G and Kaufmann P: The apoptosis cascade – morphological and immunohistochemical methods for its visualization. Anat Embryol 200: 1-18,1999.
  27. Herr I and Debatin KM: Cellular stress response and apoptosis in cancer therapy. Blood 98: 2603-2614, 2001.
  28. Debatin KM: Anticancer drugs, programmed cell death and the immune system: defining new roles in an old play. J Natl Cancer Inst 89: 750-753, 1997.
  29. Jagtap, P.; Szabo, C. Poly. (ADP-ribose) polymerase and the therapeutic effects of its inhibitors. Nat. Rev. Drug Discov. 2005, 4, 421–440.

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IssueVol 16, No 2 (2017) QRcode
SectionOriginal Article(s)
Keywords
Apoptosis FK506 Immunosuppressive Lymphocyte Proliferation Rapamycin Silymarin
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How to Cite
1.
Almasi E, Gharagozloo M, Eskandari N, Almasi A, Sabzghabaee AM. Inhibition of Apoptosis and Proliferation in T Cells by Immunosuppressive Silymarine. Iran J Allergy Asthma Immunol. 2017;16(2):107-119.