Quorum Quenching and Antimicrobial Activity of Goldenseal (Hydrastis canadensis) against Methicillin-Resistant Staphylococcus aureus (MRSA)

Abstract

The popular herbal remedy goldenseal (Hydrastis canadensis L.) is traditionally used to treat skin infections. With this study, we show activity of H. canadensis extracts in vitro against methicillin-resistant Staphylococcus aureus (MRSA). An extract from H. canadensis leaves demonstrated more potent antimicrobial activity than the alkaloid berberine alone (MICs of 75 µg/mL and 150 µg/mL, respectively). LC-MS detected alkaloids and efflux-pump inhibitory flavonoids in the extract, and the latter may explain the enhanced efficacy of the extract compared to berberine alone. We also show evidence of anti-virulence activity as a second mechanism by which H. canadensis acts against S. aureus. The H. canadensis leaf extract (but not the isolated alkaloids berberine, hydrastine, and canadine) demonstrated quorum quenching activity against several clinically relevant MRSA isolates (USA300 strains). Our data suggest that this occurs by attenuation of signal transduction through the AgrCA two-component system. Consistent with this observation, the extract inhibited toxin production by MRSA and prevented damage by MRSA to keratinocyte cells in vitro. Collectively, our results show that H. canadensis leaf extracts possess a mixture of constituents that act against MRSA via several different mechanisms. These findings lend support for the traditional application of crude H. canadensis extracts in the prevention of infection.

Key words

Hydrastis canadensis - Ranunculaceae - quorum quenching - berberine - methicillin-resistant Staphylococcus aureus - anti-virulence

• References

• 1 Klevens RM, Morrison MA, Nadle J, Petit S, Gershman K, Ray S, Harrison LH, Lynfield R, Dumyati G, Townes JM, Craig AS, Zell ER, Fosheim GE, McDougal LK, Carey RB, Fridkin SK. Invasive methicillin-resistant Staphylococcus aureus infections in the United States. JAMA 2007; 298: 1763-1771
• 2 Roberts RR, Hota B, Ahmad I, Scott D, Foster SD, Abbasi F, Schabowski S, Kampe LM, Ciavarella GG, Supino M, Naples J, Cordell R, Levy SB, Weinstein RA. Hospital and societal costs of antimicrobial-resistant infections in a chicago teaching hospital: implications for antibiotic stewardship. CID 2009; 49: 1175-1184
• 3 Waldor MK. Disarming pathogens – a new approach for antibiotic development. New Engl J Med 2006; 354: 296-297
• 4 Moran GJ, Krishnadasan A, Gorwitz RJ, Fosheim GE, McDougal LK, Carey RB, Talan DA. Methicillin-resistant S. aureus infections among patients in the emergency department. N Engl J Med 2006; 355: 666-674
• 5 Thoendel M, Kavanaugh JS, Flack CE, Horswill AR. Peptide signaling in the Staphylococci. Chem Rev 2011; 111: 117-151
• 6 Antunes LCM, Ferreira RBR, Buckner MMC, Finlay BB. Quorum sensing in bacterial virulence. Microbiology 2010; 156: 2271-2282
• 7 Mayville P, Ji G, Beavis R, Yang H, Goger M, Novick RP, Muir TW. Structure-activity analysis of synthetic autoinducing thiolactone peptides from Staphylococcus aureus responsible for virulence. Proc Natl Acad Sci USA 1999; 96: 1218-1223
• 8 Park J, Jagasia R, Kaufmann GF, Mathison JC, Ruiz DI, Moss JA, Meijler MM, Ulevitch RJ, Janda KD. Infection control by antibody disruption of bacterial quorum sensing signaling. Chem Biol 2007; 14: 1119-1127
• 9 Blumenthal M. Herb market levels after five years of boom. Herbalgram 1999; 47: 64-65
• 10 Scazzocchio F, Cometa MF, Tomassini L, Palmery M. Antibacterial activity of Hydrastis canadensis extract and its major isolated alkaloids. Planta Med 2001; 67: 561-564
• 11 Jung HA, Beavers R, Parekh H, Coulburn J, Wu CD. Goldenseal (Hydrastis canadensis) leaves possess antimicrobial activity against oral pathogens. J Dent Res 2006; 85: 475
• 12 Mahady GB, Pendland SL, 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
• 13 Villinski JR, Dumas ER, Chai HB, Pezzuto JM, Angerhofer CK, Gafner S. Antibacterial activity and alkaloid content of Berberis thunbergii, Berberis vulgaris and Hydrastis canadensis . Pharm Biol 2003; 41: 551-557
• 14 Yu HH, Kim KJ, Cha JD, Kim HK, Lee YE, Choi NY, You YO. Antimicrobial activity of berberine alone and in combination with ampicillin or oxacillin against methicillin-resistant Staphylococcus aureus . J Med Food 2005; 8: 454-461
• 15 Stermitz FR, Lorenz P, Tawara JN, Zenewicz LA, Lewis K. Synergy in a medicinal plant: antimicrobial action of berberine potentiated by 5′-methoxyhydnocarpin, a multidrug pump inhibitor. Proc Nat Acad Sci USA 2000; 97: 1433-1437
• 16 Stermitz FR, Tawara-Matsuda J, Lorenz P, Mueller P, Zenewicz LA, 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
• 17 Ettefagh KA, Burns JT, Junio HA, Kaatz GW, Cech NB. Goldenseal (Hydrastis canadensis L.) extracts synergistically enhance the antibacterial activity of berberine via efflux pump inhibition. Planta Med 2010; 77: 835-840
• 18 Upton R. Goldenseal root Hydrastis canadensis: standards of analysis, quality control, and therapeutics. Santa Cruz: American Herbal Pharmacopoeia; 2001
• 19 Moore M. Herb formulas for clinic and home. Albuquerque, NM: Southwest School of Botanical Medicine; 1995
• 20 Seybold U, Kourbatova EV, Johnson JG, Halvosa SJ, Wang YF, King MD, Ray SM, Blumberg HM. Emergence of community-associated methicillin-resistant Staphylococcus aureus USA300 genotype as a major cause of health care-associated blood stream infections. Clin Infect Dis 2006; 42: 647-656
• 21 Highlander SK, Hultén KG, Qin X, Jiang H, Yerrapragada S, Mason EO, Shang Y, Williams TM, Fortunov RM, Liu Y, Igboeli O, Petrosino J, Tirumalai M, Uzman A, Fox GE, Cardenas AM, Muzny DM, Hemphill L, Ding Y, Dugan S, Blyth PR, Buhay CJ, Dinh HH, Hawes AC, Holder M, Kovar CL, Lee SL, Liu W, Nazareth LV, Wang Q, Zhou J, Kaplan SL, Weinstock GM. Subtle genetic changes enhance virulence of methicillin resistant and sensitive Staphylococcus aureus . BMC Microbiol 2007; 7: 99
• 22 Lauderdale KJ, Boles BR, Cheung AL, Horswill AR. Interconnections between sigma B, agr, and proteolytic activity in Staphylococcus aureus biofilm maturation. Infect Immun 2009; 77: 1623-1635
• 23 Boles BR, Thoendel M, Roth AJ, Horswill AR. Identification of genes involved in polysaccharide-independent Staphylococcus aureus biofilm formation. PloS ONE 2010; 5: e10146
• 24 Malone CL, Boles BR, Horswill AR. Biosynthesis of Staphylococcus aureus autoinducing peptides by using the synechocystis DnaB mini-intein. Appl Environ Microbiol 2007; 73: 6036
• 25 Kirchdoerfer RN, Garner AL, Flack CE, Mee JM, Horswill AR, Janda KD, Kaufmann GF, Wilson IA. Structural basis for ligand recognition and discrimination of a quorum quenching antibody. J Biol Chem 2011; 286: 17351-17358
• 26 Kennedy AD, Wardenburg JB, Gardner DJ, Long D, Whitney AR, Braughton KR, Schneewind O, DeLeo FR. Targeting of alpha-hemolysin by active or passive immunization decreases severity of USA300 skin infection in a mouse model. J Infect Dis 2010; 202: 1050-1058
• 27 Thoendel M, Horswill AR. Identification of Staphylococcus aureus AgrD residues required for autoinducing peptide biosynthesis. J Biol Chem 2009; 284: 21828-21838
• 28 Pang YY, Schwartz J, Thoendel M, Ackermann LW, Horswill AR, Nauseef WM. agr-Dependent interactions of Staphylococcus aureus USA300 with human polymorphonuclear neutrophils. J Innate Immun 2010; 2: 546-559
• 29 Kiedrowski MR, Kavanaugh JS, Malone CL, Mootz JM, Voyich JM, Smeltzer MS, Bayles KW, Horswill AR. Nuclease modulates biofilm formation in community-associated methicillin-resistant Staphylococcus aureus . PLoS ONE 2011; 6: e26714
• 30 Junio HA, Sy-Cordero AA, Ettefagh KA, Burns JT, Micko KT, Graf TN, Richter SJ, Cannon RE, Oberlies NH, Cech NB. Synergy directed fractionation of botanical medicines: a case study with Goldenseal (Hydrastis canadensis). J Nat Prod 2011; 74: 1621-1629
• 31 Cech RA. Making plant medicine. Williams: Horizon Herbs; 2000: 276
• 32 Methods for dilution antimicrobial susceptibility tests for bacteria that grow aerobically. 3rd. edition. Villanova, PA: National Committee for Clinical Laboratory Standards; 1993
• 33 Boukamp P, Petrussevska RT, Breitkreutz D, Hornung J, Markham A, Fusenig NE. Normal keratinization in a spontaneously immortalized aneuploid human keratinocyte cell line. J Cell Biol 1988; 106: 761-771
• 34 Amin AH, Subbaiah TV, Abbasi KM. Berberine sulfate: antimicrobial activity, bioassay, and mode of action. Can J Microbiol 1969; 15: 1067-1076
• 35 Yu HH, Kim KJ, Cha JD, Kim HK, Lee YE, Choi NY, You YO. Antimicrobial activity of berberine alone and in combination with ampicillin or oxacillin against methicillin-resistant Staphylococcus aureus . J Med Food 2005; 8: 454-461
• 36 Coulburn J, Chin YW, Parekh H, Yildiz S, Beaver R, Kinghorn AD, Wu CD. Oral antimicrobial compounds from Hydrastis canadensis L. (Goldenseal) leaves. J Dent Res 2007; 86: 2156
• 37 McGraw JB, Sanders SM, Van der Voort M. Distribution and abundance of Hydrastis canadensis L. (Ranunculaceae) and Panax quinquefolius L. (Araliaceae) in the central Appalachian region. J Torrey Bot Soc 2003; 130: 62-69
• 38 Holtfreter S, Grumann D, Schmudde M, Nguyen HTT, Eichler P, Strommenger B, Kopron K, Kolata J, Giedrys-Kalemba S, Steinmetz I, Witte W, Bröker BM. Clonal distribution of superantigen genes in clinical Staphylococcus aureus isolates. J Clin Microbiol 2007; 45: 2669-2680