Triterpenoid Saponins from the Caryophyllaceae Family Modulate the Efflux Activity of the P-Glycoprotein in an In Vitro Model of Intestinal Barrier

Abstract

The oral bioavailability of drugs is often limited due to the presence of the P-glycoprotein, an efflux pump strongly expressed on the luminal side of the intestinal barrier. In an attempt to circumvent drug efflux, strategies consisting in the coadministration of drugs with surface-active agents have been found to be promising. In this context, the role of saponins on the intestinal permeability of a P-glycoprotein substrate was investigated. The P-glycoprotein inhibition activity of three triterpenoid saponins extracted from several plants of the Caryophyllaceae family was evaluated using an intestinal barrier model comprised of Caco-2 cell lines. The results showed a strong effect of two saponins on P-glycoprotein-mediated transport. At a concentration of 15 µM, the efflux ratio was close to 1 for both saponins, thus suggesting a total inhibition of the efflux pump in contrast to verapamil HCl, a conventional P-glycoprotein inhibitor. In addition, measurements of the transepithelial electrical resistance revealed that the integrity of the monolayers was not altered at such concentrations, thereby reducing potential adverse effects. The presence of acetylated sugars in the saponin structure could possibly facilitate interactions with the efflux pump by an ATP-dependent mechanism or by fluidization of cell membranes.

Key words

saponins - Caco-2 cells - P-glycoprotein - efflux - bioavailability

• References

• 1 Nieto Montesinos R, Béduneau A, Pellequer Y, Lamprecht A. Delivery of P-glycoprotein substrates using chemosensitizers and nanotechnology for selective and efficient therapeutic outcomes. J Control Release 2012; 161: 50-61
• 2 Lacaille-Dubois MA. Bioactive saponins from plants: recent developments. In: Yaniv Z, Bachrav U, editors Handbook of medicinal Plants. New York: Food Products Press (FPP), The Haworth Medical Press (HMP); 2005: 399-428
• 3 Onning G, Wang Q, Weström BR, Asp NG, Karlsson BW. Influence of oat saponins on intestinal permeability in vitro and in vivo in the rat. Br J Nutr 1996; 76: 141-151
• 4 Cho SY, Sim JS, Kang SS, Jeong CS, Linhardt RJ, Kim YS. Enhancement of heparin and heparin disaccharide absorption by the Phytolacca americana saponins. Arch Pharm Res 2003; 26: 1102-1108
• 5 Gaidi G, Miyamoto T, Ramezani M, Lacaille-Dubois MA. Glandulosides A–D, triterpene saponins from Acanthophyllum glandulosum . J Nat Prod 2004; 67: 1114-1118
• 6 Pertuit D, Lotfabad TB, Mitaine-Offer AC, Tomofumi M, Tanaka C, Lacaille-Dubois MA. Two new triterpene saponins from Acanthophyllum laxiusculum . Helv Chim Acta 2015; 98: 611-617
• 7 Pertuit D, Avunduk S, Mitaine-Offer AC, Miyamoto T, Tanaka C, Paululat T, Delemasure S, Dutartre P, Lacaille-Dubois MA. Triterpenoid saponins from the roots of two Gypsophila species. Phytochemistry 2014; 102: 182-188
• 8 Francis G, Kerem Z, Makkar HPS, Becker K. The biological action of saponins in animal systems: a review. Br J Nutr 2002; 88: 587-605
• 9 Hsu YL, Kuo PL, Weng TC, Yen MH, Chiang LC, Lin CC. The antiproliferative activity of saponin-enriched fraction from Bupleurum Kaoi is through Fas-dependent apoptotic pathway in human non-small cell lung cancer A549 cells. Biol Pharm Bull 2004; 27: 1112-1115
• 10 Sun Y, Cai TT, Zhou XB, Xu Q. Saikosaponin a inhibits the proliferation and activation of T cells through cell cycle arrest and induction of apoptosis. Int Immunopharmacol 2009; 9: 978-983
• 11 Narai A, Arai S, Shimizu M. Rapid decrease in transepithelial electrical resistance of human intestinal Caco-2 cell monolayers by cytotoxic membrane perturbents. Toxicol In Vitro 1997; 11: 347-354
• 12 Cho SY, Kim JS, Li H, Shim C, Linhardt RJ, Kim YS. Enhancement of paracellular transport of heparin disaccharide across Caco-2 cell monolayers. Arch Pharm Res 2002; 25: 86-92
• 13 Huang C, Xu D, Xia Q, Wang P, Rong C, Su Y. Reversal of P-glycoprotein-mediated multidrug resistance of human hepatic cancer cells by Astragaloside II. J Pharm Pharmacol 2012; 64: 1741-1750
• 14 Le Ferrec E, Chesne C, Artusson P, Brayden D, Fabre G, Gires P, Guillou F, Rousset M, Rubas W, Scarino ML. In vitro models of the intestinal barrier. The report and recommendations of ECVAM Workshop 46. European Centre for the Validation of Alternative methods. Altern Lab Anim 2001; 29: 649-668
• 15 Dudeja PK, Anderson KM, Harris JS, Buckingham L, Coon JS. Reversal of multidrug resistance phenotype by surfactants: relationship to membrane lipid fluidity. Arch Biochem Biophys 1995; 319: 309-315
• 16 Alakhova DY, Rapoport NY, Batrakova EV, Timoshin AA, Li S, Nicholls D, Alakhov VY, Kabanov AV. Differential metabolic responses to pluronic in MDR and non-MDR cells: a novel pathway for chemosensitization of drug resistant cancers. J Control Release 2010; 142: 89-100
• 17 Ozturk B, Argin S, Ozilgen M, McClements DJ. Formation and stabilization of nanoemulsion-based vitamin E delivery systems using natural surfactants: Quillaja saponin and lecithin. J Food Eng 2014; 142: 57-63
• 18 Yang Y, Leser ME, Sher AA, McClements DJ. Formation and stability of emulsions using a natural small molecule surfactant: Quillaja saponin (Q-Naturale®). Food Hydrocoll 2013; 30: 589-596
• 19 Antoine D, Pellequer Y, Tempesta C, Lorscheidt S, Kettel B, Tamaddon L, Jannin V, Demarne F, Lamprecht A, Béduneau A. Biorelevant media resistant co-culture model mimicking permeability of human intestine. Int J Pharm 2015; 481: 27-36
• 20 Béduneau A, Tempesta C, Fimbel S, Pellequer Y, Jannin V, Demarne F, Lamprecht A. A tunable Caco-2/HT29-MTX co-culture model mimicking variable permeabilities of the human intestine obtained by an original seeding procedure. Eur J Pharm Biopharm 2014; 87: 290-298
• 21 Stockert JC, Blázquez-Castro A, Cañete M, Horobin RW, Villanueva A. MTT assay for cell viability: Intracellular localization of the formazan product is in lipid droplets. Acta Histochem 2012; 114: 785-796