Browse > Article
http://dx.doi.org/10.4062/biomolther.2014.072

Silymarin Inhibits Cytokine-Stimulated Pancreatic Beta Cells by Blocking the ERK1/2 Pathway  

Kim, Eun Jeong (Department of Pharmacology, School of Medicine, Chosun University)
Kim, Jeeho (Department of Pharmacology, School of Medicine, Chosun University)
Lee, Min Young (Department of Pharmacology, School of Medicine, Chosun University)
Sudhanva, Muddenahalli Srinivasa (Department of Pharmacology, School of Medicine, Chosun University)
Devakumar, Sundaravinayagam (Department of Pharmacology, School of Medicine, Chosun University)
Jeon, Young Jin (Department of Pharmacology, School of Medicine, Chosun University)
Publication Information
Biomolecules & Therapeutics / v.22, no.4, 2014 , pp. 282-287 More about this Journal
Abstract
We show that silymarin, a polyphenolic flavonoid isolated from milk thistle (Silybum marianum), inhibits cytokine mixture (CM: TNF-${\alpha}$, IFN-${\gamma}$, and IL-$1{\beta}$)-induced production of nitric oxide (NO) in the pancreatic beta cell line MIN6N8a. Immunostaining and Western blot analysis showed that silymarin inhibits iNOS gene expression. RT-PCR showed that silymarin inhibits iNOS gene expression in a dose-dependent manner. We also showed that silymarin inhibits extracellular signal-regulated protein kinase-1 and 2 (ERK1/2) phosphorylation. A MEK1 inhibitor abrogated CM-induced nitrite production, similar to silymarin. Treatment of MIN6N8a cells with silymarin also inhibited CM-stimulated activation of NF-${\kappa}B$, which is important for iNOS transcription. Collectively, we demonstrate that silymarin inhibits NO production in pancreatic beta cells, and silymarin may represent a useful anti-diabetic agent.
Keywords
Silymarin; Beta cells; NO; iNOS; ERK1/2;
Citations & Related Records
Times Cited By KSCI : 4  (Citation Analysis)
연도 인용수 순위
1 Zhao, J., Sharma, Y. and Agarwal, R. (1999) Signifi cant inhibition by the flavonoid antioxidant silymarin against 12-O-tetradecanoylphorbol 13-acetate-caused modulation of antioxidant and inflammatory enzymes, and cyclooxygenase 2 and interleukin-1alpha expression in SENCAR mouse epidermis: implications in the prevention of stage I tumor promotion. Mol. Carcinog. 26, 321-333.   DOI   ScienceOn
2 Pliskova, M., Vondracek, J., Kren, V., Gazak, R., Sedmera, P., Walterova, D., Psotova, J., Simanek, V. and Machala, M. (2005) Effects of silymarin flavonolignans and synthetic silybin derivatives on estrogen and aryl hydrocarbon receptor activation. Toxicology 215, 80-89.   DOI
3 Soto, C., Mena, R., Luna, J., Cerbon, M., Larrieta, E., Vital, P., Uria, E., Sanchez, M., Recoba, R., Barron, H., Favari, L. and Lara, A. (2004) Silymarin induces recovery of pancreatic function after alloxan damage in rats. Life Sci. 75, 2167-2180.   DOI
4 Soto, C., Raya, L., Juarez, J., Perez, J. and Gonzalez, I. (2014) Effect of Silymarin in Pdx-1 expression and the proliferation of pancreatic beta-cells in a pancreatectomy model. Phytomedicine 21, 233-239.   DOI
5 Takamura, T., Kato, I., Kimura, N., Nakazawa, T., Yonekura, H., Takasawa, S. and Okamoto, H. (1998) Transgenic mice overexpressing type 2 nitric-oxide synthase in pancreatic beta cells develop insulin-dependent diabetes without insulitis. J. Biol. Chem. 273, 2493-2496.   DOI
6 Valenzuela, A. and Garrido, A. (1994) Biochemical bases of the pharmacological action of the flavonoid silymarin and of its structural isomer silibinin. Biol. Res. 27, 105-112.
7 Welsh, N. and Sandler, S. (1992) Interleukin-1 beta induces nitric oxide production and inhibits the activity of aconitase without decreasing glucose oxidation rates in isolated mouse pancreatic islets. Biochem. Biophys. Res. Commun. 182, 333-340.   DOI   ScienceOn
8 Xie, H., Chiles, T. C. and Rothstein, T. L. (1993) Induction of CREB activity via the surface Ig receptor of B cells. J. Immunol. 151, 880-889.
9 Xie, Q. W., Kashiwabara, Y. and Nathan, C. (1994) Role of transcription factor NF-kappa B/Rel in induction of nitric oxide synthase. J. Biol. Chem. 269, 4705-4708.
10 Youn, C. K., Park, S. J., Li, M. H., Lee, M. Y., Lee, K. Y., Cha, M. J., Kim, O. H., You, H. J., Chang, I. Y., Yoon, S. P. and Jeon, Y. J. (2013) Radi cicol inhibits iNOS expression in cytokine-stimulated pancreatic beta cells. Korean J. Physiol. Pharmacol. 17, 315-320.   DOI
11 Zhang, W., Hong, R. and Tian, T. (2013) Silymarin's protective effects and possible mechanisms on alcoholic fatty liver for rats. Biomol. Ther. 21, 264-269.   DOI
12 Kvasnicka, F., Biba, B., Sevcik, R., Voldrich, M. and Kratka, J. (2003) Analysis of the active components of silymarin. J. Chromatogr. A 990, 239-245.   DOI
13 Larsen, L., Storling, J., Darville, M., Eizirik, D. L., Bonny, C., Billestrup, N. and Mandrup-Poulsen, T. (2005) Extracellular signal-regulated kinase is essential for interleukin-1-induced and nuclear factor kappaB-mediated gene expression in insulin-producing INS-1E cells. Diabetologia 48, 2582-2590.   DOI   ScienceOn
14 Li, M. H., Kothandan, G., Cho, S. J., Huong, P. T., Nan, Y. H., Lee, K. Y., Shin, S. Y., Yea, S. S. and Jeon, Y. J. (2010) Magnolol inhibits LPS-induced NF-kappaB/rel activation by blocking p38 kinase in murine macrophages. Korean J. Physiol. Pharmacol. 14, 353-358.   과학기술학회마을   DOI
15 Matsuda, T., Ferreri, K., Todorov, I., Kuroda, Y., Smith, C. V., Kandeel, F. and Mullen, Y. (2005) Silymarin protects pancreatic beta-cells against cytokine-mediated toxicity: implication of c-Jun NH2-terminal kinase and janus kinase/signal transducer and activator of transcription pathways. Endocrinology 146, 175-185.   DOI   ScienceOn
16 Mereish, K. A., Bunner, D. L., Ragland, D. R. and Creasia, D. A. (1991) Protection against microcystin-LR-induced hepatotoxicity by Silymarin: biochemistry, histopathology, and lethality. Pharm. Res. 8, 273-277.   DOI   ScienceOn
17 Eldor, R., Yeffet, A., Baum, K., Doviner, V., Amar, D., Ben-Neriah, Y., Christofori, G., Peled, A., Carel, J. C., Boitard, C., Klein, T., Serup, P., Eizirik, D. L. and Melloul, D. (2006) Conditional and specifi c NFkappaB blockade protects pancreatic beta cells from diabetogenic agents. Proc. Natl. Acad. Sci. U.S.A. 103, 5072-5077.   DOI   ScienceOn
18 Mokhtari, M. J., Motamed, N. and Shokrgozar, M. A. (2008) Evaluation of silibinin on the viability, migration and adhesion of the human prostate adenocarcinoma (PC-3) cell line. Cell Biol. Int. 32, 888-892.   DOI   ScienceOn
19 Oh, S. J., Jung, S. P., Han, J., Kim, S., Kim, J. S., Nam, S. J., Lee, J. E. and Kim, J. H. (2013) Silibinin inhibits TPA-induced cell migration and MMP-9 expression in thyroid and breast cancer cells. Oncol. Rep. 29, 1343-1348.   DOI
20 Oleson, B. J., Broniowska, K. A., Schreiber, K. H., Tarakanova, V. L. and Corbett, J. A. (2014) Nitric oxide induces ataxia telangiectasia mutated (ATM) protein-dependent gammaH2AX protein formation in pancreatic beta cells. J. Biol. Chem. 289, 11454-11464.   DOI
21 Flodstrom, M., Tyrberg, B., Eizirik, D. L. and Sandler, S. (1999) Reduced sensitivity of inducible nitric oxide synthase-deficient mice to multiple low-dose streptozotocin-induced diabetes. Diabetes 48, 706-713.   DOI
22 Green, L. C., Wagner, D. A., Glogowski, J., Skipper, P. L., Wishnok, J. S. and Tannenbaum, S. R. (1982) Analysis of nitrate, nitrite, and [15N]nitrate in biological fluids. Anal. Biochem. 126, 131-138.   DOI   ScienceOn
23 Heimberg, H., Heremans, Y., Jobin, C., Leemans, R., Cardozo, A. K., Darville, M. and Eizirik, D. L. (2001) Inhibition of cytokine-induced NF-kappaB activation by adenovirus-mediated expression of a NFkappaB super-repressor prevents beta-cell apoptosis. Diabetes 50, 2219-2224.   DOI   ScienceOn
24 Kang, J. S., Jeon, Y. J., Kim, H. M., Han, S. H. and Yang, K. H. (2002) Inhibition of inducible nitric-oxide synthase expression by silymarin in lipopolysaccharide-stimulated macrophages. J. Pharmacol. Exp. Ther. 302, 138-144.   DOI   ScienceOn
25 Hogan, F. S., Krishnegowda, N. K., Mikhailova, M. and Kahlenberg, M. S. (2007). Flavonoid, silibinin, inhibits proliferation and promotes cell-cycle arrest of human colon cancer. J. Surg. Res. 143, 58-65.   DOI   ScienceOn
26 Huong, P. T., Lee, M. Y., Lee, K. Y., Chang, I. Y., Lee, S. K., Yoon, S. P., Lee, D. C. and Jeon, Y. J. (2012) Synergistic induction of iNOS by IFN-gamma and glycoprotein isolated from Dioscorea batatas. Korean J. Physiol. Pharmacol. 16, 431-436.   과학기술학회마을   DOI   ScienceOn
27 Jeon, Y. J., Yang, K. H., Pulaski, J. T. and Kaminski, N. E. (1996) Attenuation of inducible nitric oxide synthase gene expression by delta 9-tetrahydrocannabinol is mediated through the inhibition of nuclear factor- kappa B/Rel activation. Mol. Pharmacol. 50, 334-341.
28 Kim, S., Choi, J. H., Lim, H. I., Lee, S. K., Kim, W. W., Kim, J. S., Kim, J. H., Choe, J. H., Yang, J. H., Nam, S. J. and Lee, J. E. (2009a) Silibinin prevents TPA-induced MMP-9 expression and VEGF secretion by inactivation of the Raf/MEK/ERK pathway in MCF-7 human breast cancer cells. Phytomedicine 16, 573-580.   DOI   ScienceOn
29 Kim, S., Choi, M. G., Lee, H. S., Lee, S. K., Kim, S. H., Kim, W. W., Hur, S. M., Kim, J. H., Choe, J. H., Nam, S. J., Yang, J. H., Lee, J. E. and Kim, J. S. (2009b) Silibinin suppresses TNF-alpha-induced MMP-9 expression in gastric cancer cells through inhibition of the MAPK pathway. Molecules 14, 4300-4311.   DOI   ScienceOn
30 Baker, M. S., Chen, X., Cao, X. C. and Kaufman, D. B. (2001) Expression of a dominant negative inhibitor of NF-kappaB protects MIN6 beta-cells from cytokine-induced apoptosis. J. Surg. Res. 97, 117-122.   DOI
31 Corbett, J. A., Wang, J. L., Hughes, J. H., Wolf, B. A., Sweetland, M. A., Lancaster, J. R., Jr. and McDaniel, M. L. (1992) Nitric oxide and cyclic GMP formation induced by interleukin 1 beta in islets of Langerhans. Evidence for an effector role of nitric oxide in islet dysfunction. Biochem. J. 287 ( Pt 1), 229-235.   DOI
32 Bektur, N. E., Sahin, E., Baycu, C. and Unver, G. (2013) Protective effects of silymarin against acetaminophen-induced hepatotoxicity and nephrotoxicity in mice. Toxicol. Ind. Health, 1-12.
33 Broniowska, K. A., Oleson, B. J. and Corbett, J. A. (2014) beta-cell responses to nitric oxide. Vitam. Horm. 95, 299-322.   DOI
34 Cetkovic-Cvrlje, M. and Eizirik, D. L. (1994) TNF-alpha and IFN-gamma potentiate the deleterious effects of IL-1 beta on mouse pancreatic islets mainly via generation of nitric oxide. Cytokine 6, 399-406.   DOI   ScienceOn
35 Cristofalo, R., Bannwart-Castro, C. F., Magalhaes, C. G., Borges, V. T., Peracoli, J. C., Witkin, S. S. and Peracoli, M. T. (2013) Silibinin atte nuates oxidative metabolism and cytokine production by monocytes from preeclamptic women. Free Radic. Res. 47, 268-275.   DOI
36 Darville, M. I. and Eizirik, D. L. (1998) Regulation by cytokines of the inducible nitric oxide synthase promoter in insulin-producing cells. Diabetologia 41, 1101-1108.   DOI   ScienceOn
37 Dunnett, M. (1955) A multiple comparison procedure for comparing several treatments with a control. J. Am. Stat. Assoc. 50, 1096-1121.   DOI   ScienceOn
38 Letteron, P., Labbe, G., Degott, C., Berson, A., Fromenty, B., Delaforge, M., Larrey, D. and Pessayre, D. (1990) 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. 39, 2027-2034.   DOI   ScienceOn
39 Yamada, K., Otabe, S., Inada, C., Takane, N. and Nonaka, K. (1993) Nitric oxide and nitric oxide synthase mRNA induction in mouse islet cells by interferon-gamma plus tumor necrosis factor-alpha. Biochem. Biophys. Res. Commun. 197, 22-27.   DOI   ScienceOn