In Vitro and Ex Vivo Effects of Silymarin Derivatives on Proinflammatory Cytokine Secretion Using a Murine Colonic Strip Model

Abstract

Silymarin has anti-inflammatory properties and documented anti-colitis activity. Our prior study determined that in vitro treatment with certain extracted fractions of silymarin inhibited stimulated proinflammatory cytokine secretion from cell lines relevant to colitis. In this study, colitis was induced in mice by giving dextran sulfate sodium drinking water for 6 days. The ex vivo effects of crude silymarin extract, two different silymarin fractions, as well as commercially derived silibinin and isosilibinin were examined by determining the secretion of MIP-2, TNF-α, and IL-17 in cell culture media from colonic strips. Further, the effects of silymarin-derived treatments on IL-8 and TNF-α secretion induced by the colitis supernatant was characterized with HT-29 colonic epithelial and RAW 264.7 macrophage cell lines. Prominent inhibition of MIP-2 and TNF-α secretion from colonic strips of mice with/without dextran sulfate sodium-induced colitis was observed with various silymarin treatments. Further, inhibition of dual (IL-23+IL-1β) cytokine-stimulated secretion of IL-17 from colonic strips of mice was found with certain silymarin treatments. Significant attenuation of TNF-α secretion from colitis supernatant-stimulated RAW 264.7 cells was observed for crude silymarin extract and isosilibinin treatments. Finally, inhibition of IL-8 secretion from the colitis supernatant-stimulated HT29 colonic epithelial cell line was found with isosilibinin. These results contribute to the identification of silymarin-derived flavonoligans with optimal anti-inflammatory properties for further testing in colitis models.

Key words

silymarin - silibinin - isosilibinin - anti-inflammatory - cytokines - inflammatory bowel disease

• References

• 1 Fitzpatrick LR. Novel pharmacological approaches for inflammatory bowel disease: Targeting key intracellular pathways and the IL-23/IL-17 axis. Int J Inflam 2012; 2012: 389404
• 2 Zhu HJ, Brinda BJ, Chavin KD, Bernstein HJ, Patrick KS, Markowitz JS. An assessment of pharmacokinetics and antioxidant activity of free silymarin flavonoligans in health volunteers: A dose escalation study. Drug Metab Dispos 2013; 41: 1479-1485
• 3 Biijak M. Silybin, a major bioactive component of milk thistle (Silybum marianum L Gaernt.)-chemistry, bioavailability and metabolism. Molecules 2017; 22: E1942
• 4 Abenavoli L, Capasso R, Milic N, Capasso F. Milk thistle in liver diseases: Past, present, future. Phytother Res 2010; 24: 1423-1432
• 5 Neha Jaggi AS, Singh N. Silymarin and Its Role in Chronic Diseases. Adv Exp Med Biol 2016; 929: 25-44
• 6 Polachi N, Bai G, Li T, Chu Y, Wang X, Li S, Gu N, Wu J, Li W, Zhang Y, Zhou S, Sun H, Liu C. Modulatory effects of silibinin in various cell signaling pathways against liver disorders and cancer. Eur J Med Chem 2016; 123: 577-595
• 7 Scambia G, De Vincenzo R, Ranelletti FO, Panici PB, Ferrandina G, D'Agostino G, Fattorossi A, Bombardelli E, Mancuso S. Antiproliferative effect of silybin on gynaecological malignancies: Synergism with cisplatin and doxorubicin. Eur J Cancer 1996; 32A: 877-882
• 8 Rastegarpanah M, Malekzadeh R, Vahedi H, Mohammadi M, Elahi E, Chaharmahali M, Safarnavadeh T, Abdollahi M. A randomized, double blinded, placebo-controlled clinical trial of silymarin in ulcerative colitis. Chin J Integr Med 2015; 21: 902-906
• 9 Talbott G, Woldemariam T, Fitzpatrick LR. Effects of silymarin fractions on pro-inflammatory cytokine secretion from macrophage and colonic epithelial cell lines. Pharm Pharmacol Int J 2017; 5: 00128
• 10 Fitzpatrick LR, Stonesifer E, Small JS, Liby KT. The synthetic triterpenoid (CDDO-Im) inhibits STAT3, as well as IL-17, and improves DSS-induced colitis in mice. Inflammopharmacology 2014; 22: 341-349
• 11 Ohstuka Y, Sanderson IR. Dextran sulfate-sodium induced inflammation is enhanced by intestinal epithelial cell chemokine expression in mice. Pediatr Res 2003; 53: 143-147
• 12 Kleinscheck MA, Boniface K, Sadevoka S, Grein J, Murphy EE, Turner SP, Raskin L, Desai B, Faubion WA, de Waal Malefyt R, Pierce RH, McClanahan T, Kastelein RA. Circulating and gut resident human Th17 cells express CD161 and promote intestinal inflammation. J Exp Med 2009; 206: 525-534
• 13 Coccia M, Harrison OJ, Schiering C, Asquith MJ, Becher B, Powrie F, Maloy KJ. IL-1β mediates chronic intestinal inflammation by promoting the accumulation of IL-17A secreting innate lymphoid cells and CD4(+) Th17cells. J Exp Med 2012; 209: 1595-1609
• 14 Withers DR, Hepworth MR, Wang X, Mackley EC, Halford EE, Dutton EE, Marriott CL, Brucklacher-Waldert V, Veldhoen M, Kelsen J, Baldassano RN, Sonnenberg GF. Transient inhibition of ROR-γT therapeutically limits intestinal inflammation by reducing TH17 cells and preserving group 3 innate lymphoid cells. Nat Med 2016; 22: 319-323
• 15 Fitzpatrick LR, Talbott G, Buhr CA, Alton G, Zapf J. Ex vivo effects of ROR-gamma T inhibitors on proinflammatory cytokine secretion from colonic strips of mice with DSS-induced colitis. Gastroenterology 2018; 154: S1007-S1008
• 16 Fitzpatrick LR, Wang J, Le T. In vitro and in vivo effects of gliotoxin, a fungal metabolite: efficacy against dextran sodium sulfate-induced colitis in rats. Dig Dis Sci 2000; 45: 2327-2336
• 17 Fitzpatrick LR, Wang T, Le T. Caffeic acid phenethyl ester, an inhibitor of nuclear factor-kappaB, attenuates bacterial peptidoglycan polysaccharide-induced colitis in rats. J Pharmacol Exp Ther 2001; 299: 915-920
• 18 Fitzpatrick LR, Talbott G, Woldemariam T. Effects of silymarin fractions on pro-inflammatory cytokine secretion from macrophage and colonic epithelial cell lines. Gastroenterology 2016; 150: S376-S377
• 19 Surai PF. Silymarin as a Natural Antioxidant: An Overview of the Current Evidence and Perspective. Antioxidants (Basel) 2015; 4: 204-247
• 20 Raina K, Agarwal C, Agarwal R. Effect of silibinin in human colorectal cancer cells: targeting the activation of NF-κB signaling. Mol Carcinog 2013; 52: 195-206
• 21 Esmaily H, Hosseini-Tabatabaei A, Rahimian R, Khorasani R, Baeeri M, Barazesh-Morgani A, Yesa N, Khademi Y, Abdollahi M. On the benefits of silymarin in murine colitis by improving balance of destructive cytokines and reduction of toxic stress in the bowel cells. Cent Eur J Biol 2009; 4: 204-213
• 22 Esmaily H, Vaziri-Bami A, Miroliaee AE, Baeeri M, Abdollahi M. The correlation between NF-κB inhibition and disease activity by coadministration of silibinin and ursodeoxycholic acid in experimental colitis. Fundam Clin Pharmacol 2011; 25: 723-733
• 23 Woldemariam TZ, Malekakhlagh A, Bett C, Pearson D. Evaluation of the antitumor activity of selected herbs and spices. J Pharm Sci Pharmacol 2014; 1: 296-303