Goldenseal (Hydrastis canadensis L.) Extracts Synergistically Enhance the Antibacterial Activity of Berberine via Efflux Pump Inhibition

Abstract

Goldenseal (Hydrastis canadensis L.) is used to combat inflammation and infection. Its antibacterial activity in vitro has been attributed to its alkaloids, the most abundant of which is berberine. The goal of these studies was to compare the composition, antibacterial activity, and efflux pump inhibitory activity of ethanolic extracts prepared from roots and aerial portions of H. canadensis. Ethanolic extracts were prepared separately from roots and aerial portions of six H. canadensis plants. Extracts were analyzed for alkaloid concentration using LC‐MS and tested for antimicrobial activity against Staphylococcus aureus (NCTC 8325-4) and for inhibition of ethidium bromide efflux. Synergistic antibacterial activity was observed between the aerial extract (FIC 0.375) and to a lesser extent the root extract (FIC 0.750) and berberine. The aerial extract inhibited ethidium bromide efflux from wild-type S. aureus but had no effect on the expulsion of this compound from an isogenic derivative deleted for norA. Our studies indicate that the roots of H. canadensis contain higher levels of alkaloids than the aerial portions, but the aerial portions synergize with berberine more significantly than the roots. Furthermore, extracts from the aerial portions of H. canadensis contain efflux pump inhibitors, while efflux pump inhibitory activity was not observed for the root extract. The three most abundant H. canadensis alkaloids, berberine, hydrastine, and canadine, are not responsible for the efflux pump inhibitory activity of the extracts from H. canadensis aerial portions.

Key words

Hydrastis canadensis - Ranunculaceae - synergy - alkaloid - berberine - Staphylococcus aureus

References

• 1 Klevens R M, Morrison M A, Nadle J, Petit S, Gershman K, Ray S, Harrison L H, Lynfield R, Dumyati G, Townes J M, Craig A S, Zell E R, Fosheim G E, McDougal L K, Carey R B, Fridkin S K. Invasive methicillin-resistant Staphylococcus aureus infections in the United States.  JAMA. 2007;  298 1763-1771
• 2 Spelman K, Duke J A, Bogenschutz-Godwin M J. The synergy principle at work with plants, pathogens, insects, herbivores and humans. Cseke L, Kirakosyan A, Kaufman PB Natural products from plants, 2nd edition. Boca Raton; CRC Taylor and Francis 2006: 475-501
• 3 Wagner H, Ulrich-Merzenich G. Synergy research: approaching a new generation of phytopharmaceuticals.  Phytomedicine. 2009;  16 97-110
• 4 Bannerman J E. Goldenseal in world trade: pressures and potentials.  Herbalgram. 1997;  2 51-53
• 5 Sinclair A, Catling P M. Cultivating the increasingly popular medicinal plant, goldenseal: Review and update.  Am J Altern Agric. 2001;  16 131-140
• 6 Blumenthal M. Herb market levels after five years of boom.  Herbalgram. 1999;  47 64-65
• 7 Hwang B Y, Roberts S K, Chadwick L, Wu C D, Kinghorn D A. Antimicrobial constitutents from goldenseal (the rhizomes of Hydrastis canadensis) against selected oral pathogens.  Planta Med. 2003;  69 623-627
• 8 Khosla P K, Neeraj V I, Gupta S K, Satpathy G. Berberine, a potential drug for trachoma.  Rev Int Trach Pathol Ocul Trop Subtrop Sante Publique. 1992;  69 147-165
• 9 Knight S E. Goldenseal (Hydrastis canadensis) versus penicillin: a comparison of effects on Staphylococcus aureus, Streptococcus pyogenes, and Pseudomonas aeruginosa.  Bios. 1999;  70 3-10
• 10 Mahady G B, Pendland S L, Stoia A, Chadwick L. In vitro suceptibility of Helicobacter pylori to isoquinoline alkaloids from Sanguinaria canadensis and Hydrastis canadensis.  Phytother Res. 2003;  17 217-221
• 11 Scazzocchino F, Cometa M F, Tomassini L, Palmery M. Antibacterial activity of Hydrastis canadensis extract and its major isolated alkaloids.  Planta Med. 2001;  67 561-564
• 12 Chadwick L R, Wu C D, Kinghorn A D. Isolation of alkaloids from goldenseal (Hydrastis canadensis rhizomes) using pH-zone refining countercurrent chromatography.  J Liq Chromatogr Relat Technol. 2001;  24 2445-2453
• 13 Villinski J R, Dumas E R, Chai H B, Pezzuto J M, Angerhofer C K, Gafner S. Antibacterial activity and alkaloid content of Berberis thunbergii, Berberis vulgaris and Hydrastis canadensis.  Pharm Biol. 2003;  41 551-557
• 14 Gentry E J, Jampani H B, Keshavarz-Shokri A, Morton M D, Velde D V, Telikepalli H, Mitscher L A, Shawar R, Humble D, Baker W. Antitubercular natural products: Berberine from the roots of commercial Hydrastis canadensis powder. Isolation of inactive 8-oxotetrahydrothalifendine, canadine, b-hydrastine, and two new quinic acid esters, hycandinic acid esters-1 and 2.  J Nat Prod. 1998;  61 1187-1193
• 15 Nikaido H. Multiple antibiotic resistance and efflux.  Curr Opin Microbiol. 1998;  1 516-523
• 16 Webber M A, Piddock L J V. The importance of efflux pumps in bacterial antibiotic resistance.  J Antimicrob Chemother. 2003;  51 9-11
• 17 Lewis K, Ausubel F M. Prospects for plant-derived antibacterials.  Nat Biotechnol. 2006;  24 1504-1507
• 18 Novick R. Properties of cryptic high-frequency transducing phage of Staphylococcus aureus.  Virology. 1967;  33 155-166
• 19 Price C T, Kaatz G W, Gustafson J E. The multidrug efflux pump NorA is not required for salicylate induced reduction in drug accumulation by Staphylococcus aureus.  Int J Antimicrob Agents. 2002;  20 206-213
• 20 Cech R A. Making plant medicine. Williams; Horizon Herbs 2000: 276
• 21 No author. Methods for dilution antimicrobial susceptibility tests for bacteria that grow aerobically; approved standard, Vol. 26, 7th edition. Wayne; Clinical and Laboratory Standards Institute 2006: M7-A7
• 22 Eliopolous G M, Moellering R C. Antimicrobial combinations. Lorian V Antibiotics in laboratory medicine. Baltimore, MD; Williams and Wilkins 1996: 330-396
• 23 Amin A H, Subbaiah T V, Abbasi K M. Berberine sulfate: antimicrobial activity, bioassay, and mode of action.  Can J Microbiol. 1969;  15 1067-1076
• 24 Yu H H, Kim K J, Cha J D, Kim H K, Lee Y E, Choi N Y, You Y O. Antimicrobial activity of berberine alone and in combination with ampicillin or oxacillin against methicillin-resistant Staphylococcus aureus.  J Med Food. 2005;  8 454-461
• 25 Scazzocchio F, Cometa M F, Tomassini L, Palmery M. Antibacterial activity of Hydrastis canadensis extract and its major isolated alkaloids.  Planta Med. 2001;  67 561-564
• 26 Monkovic I, Spenser I D. Biosynthesis of berberine and berverastine.  Can J Chem. 1965;  43 2017-2026
• 27 Upton R. Goldenseal root Hydrastis canadensis: Standards of analysis, quality control, and therapeutics. Santa Cruz; American Herbal Pharmacopoeia 2001
• 28 Odds F C. Synergy, antagonism, and what the chequerboard puts between them.  J Antimicrob Chemother. 2003;  52 1
• 29 Stermitz F R, Tawara-Matsuda J, Lorenz P, Mueller P, Zenewicz L A, Lewis K. 5′Methoxyhydnocarpin-D and phenophorbide A: Berberis species components that potentiate berberine growth inhibition of resistant Staphylococcus aureus.  J Nat Prod. 2000;  63 1146-1149
• 30 Wang D, Yu L, Xiang H, Fan J, He L, Guo N, Feng H, Deng X. Global transcriptional profiles of Staphylococcus aureus treated with berberine chloride.  FEMS Microbiol Lett. 2008;  279 217-225
• 31 Yu H-H, Kim K-J, Cha J-D, Kim H-K, Lee Y-E, Choi N-Y, You Y-O. Antimicrobial activity of berberine alone and in combination with ampicillin or oxacillin against methicillin-resistant Staphylococcus aureus.  J Med Food. 2005;  8 454-461
• 32 Tegos G, Stermitz F R, Lomovskaya O, Lewis K. Multidrug pump inhibitors uncover remarkable activity of plant antimicrobials.  Antimicrob Agents Chemother. 2002;  46 3133-3141
• 33 Stermitz F R, Lorenz P, Tawara J N, Zenewicz L A, Lewis K. Synergy in a medicinal plant: antimicrobial action of berberine potentiated by 5′-methoxyhydnocarpin, a multidrug pump inhibitor.  Proc Nat Acad Sci. 2000;  97 1433-1437