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Antibacterial Activity of Sophoraflavanone G Isolated from the Roots of Sophora Flavescens and Red Ginseng Extract

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Antibacterial Activity of Sophoraflavanone G Isolated from the Roots of Sophora Flavescens and Red Ginseng Extract International Journal of Medicinal Plants. Photon 106 (2014) 418-429 https://sites.google.com/site/photonfoundationorganization/home/international-journal-of-medicinal-plants Original Research Article. 6672-4384 International Journal of Medicinal Plants Ph ton Antibacterial activity of Sophoraflavanone G isolated from the roots of Sophora flavescens and Red Ginseng Extract Jeong-Dan Cha a, Mi-Ran Jeong a, Seung-Mi Hwang a, Ji-Ye Lim a, Jea-Ran Kang a, Tae-Bum Lee b, Min-Jung Lee b, Hoo-Kil Jung c, Joo-Hee Lee c, Kyung-Min Choi a* a Institute of Jinan Red Ginseng, 520-9 Banwol-ri, Jinan-eup, Jinan-gun, Jeollabuk-do, 567-801 South Korea b Gochang Black Raspberry Research Institute, 558 Bokbunja-ro, Buan-myun, Gochang-gun, Jeollabuk-do, South Korea c Imsil Research Institute of Cheese Science, 50 Doin 2-gil, Seongsu-myeon, Imsil-gun, Jeollabuk-do 566-881, South Korea The authors receive Thomas Edison Award-2014 in Lee M.J. Medicinal Plants for Inspiration and Knowledge Researcher Distribution among young research scholars. Email: [email protected] Article history: Jung H.K. Received: 20 June, 2013 President Accepted: 26 June , 2013 Email: [email protected] Available online: 30 January, 2014 Lee J.H. Abbreviations: Researcher MICs: Minimum Inhibitory Concentrations, MBCs: Email: [email protected] Minimum Bactericidal Concentrations, CFU: Colony Forming Unit, FIC index: Fractional Inhibitory Abstract Concentration, FBC index: Fractional Bactericidal The objective of this study was to investigate the Concentration index, RGE: Red Ginseng Extract antibacterial activities of sophoraflavanone G from Sophora flavescens and Red Ginseng Extract Keywords: Sophora flavescens , Sophoraflavanone G, Red Ginseng (RGE) in combination with two antimicrobial agents extract, antibacterial activity, Checker board method, time against oral bacteria. The effects of kill method, synergic effect sophoraflavanone G + RGE combination was synergistic against all tested oral bacteria by FICI Corresponding Author: <0.5, except S. sanguinis and A. Choi K.M.* Ph.D actinomycetemcomitans and against S. sanguinis, Lab Head S. ratti, S. anginosus, F. nucleatum, P. intermedia, Email: [email protected] and P. gingivalis (FBCI <0.5). In particular, the Phone: +82634320913 Fax No: +82634320910 MICs/MBCs were reduced to one half ~ one sixteenth as a result of the combination for all Cha J.D. Ph.D bacteria. Furthermore, a time-kill study showed that Email: [email protected] the growth of the tested bacteria was completely attenuated after 2-6 h of treatment with the 1/2 MIC Jeong M.R. Ph.D of sophoraflavanone G and RGE, regardless of Email: [email protected] whether it was administered alone or combination or with ampicillin or gentamicin. The in vitro data Hwang S.M. Researcher suggest that sophoraflavanone G combined with Email: [email protected] RGE or other antibiotics may be microbiologically beneficial and not antagonistic. Lim J.Y. Researcher Citation: Email: [email protected] Cha J.D., Jeong M.R., Hwang S.M., Lim J.Y., Kang J.R., Lee T.B., Lee M.J., Jung H.K., Lee J.H., Choi K.M., 2014. Kang J.R. Antibacterial activity of Sophoraflavanone G isolated from Researcher the roots of Sophora flavescens and Red Ginseng Extract. Email: [email protected] International Journal of Medicinal Plants. Photon 106, 418-429. Lee T.B. Ph.D Email: [email protected] Ph ton 418 1. Introduction Dental caries and periodontal disease are have a radioprotective effect against radiation- prevalent worldwide. Bacteria existing in the induced double-strand breaks in DNA and dental plaque or biofilm play an important role immunomodulatory activity, as evidenced by in the development of both dental caries and its stimulation of natural killer cells (Kenarova periodontal disease (Iwano et al., 2010; et al., 1990; Seo et al., 2013). Ginsenosides Marsh, 2005). Specific bacteria are also exhibit less potent but broad-spectrum closely related to specific dental diseases, for antibacterial activity against Gram-positive and example, Streptococcus mutans and Gram-negative bacterial strains, including the Porphyromonas gingivalis are associated with clinical isolates of MRSA (Kochan et al., 2013; dental caries and periodontal disease, Sung, Lee, 2008). respectively (Iwano et al., 2010; Burton et al., 2011; Sambunjak et al., 2011). Dental caries Sophorae radix , the dried root of Sophora and periodontitis are common oral diseases flavescens AITON (Leguminosae), is an caused by bacterial infection and the important herbal medicine, which has been development of dental plaque (Marsh, 1992; traditionally employed as an antipyretic, Loesche, 2007). Several antibacterial agents analgesic, antihelminthic, and stomachic, and including, fluorides, phenol derivatives, has been shown to exert antibacterial, anti- ampicillin, erythromycin, penicillin, tetracycline, inflammatory, antioxidant, and antimalarial and vancomycin have been used widely in effects (Kuroyanagi et al., 1999; Kim et al., dentistry to inhibit bacterial growth (Levi, 2004; Hsiang et al., 2001). S. flavescens is Eusterman, 2011; Nguyen, Marqui, 2011; De also known to harbor numerous flavonoids, Poi, 2001; Emilson, 1994). However, including formononetin, kushenol E, kushenol excessive use of these chemicals can result in B, sophoraflavanone G, kushenol L, kushenol derangements of the oral and intestinal flora M, kuraridin, kurarinone, kushenol N, and and cause side effects such as microorganism kushenol F (Jin et al., 2010; DeNaeyer et al., susceptibility, vomiting, diarrhoea and tooth 2004; Ryu et al., 1960; Shen et al., 2006). staining (Schaeken et al., 1991; Ekstrand et Recently, several researchers have reported al., 1998; Feres et al., 2010). These problems that these compounds have proven effective necessitate further search for natural against tumors, arrhythmia, and antibacterial agents that are safe for humans immunodeficiency, antitumor, antioxidant, and and specific for oral pathogens. Natural antibacterial activity and thus have generated products have recently been investigated more a great deal of attention and interest (Jin et al., thoroughly as promising agents to prevent oral 2010; DeNaeyer et al., 2004; Kim et al., 2002; diseases, especially plaque-related diseases Zhang et al., 2006). We reported previously such as dental caries (Feldman et al., 2011; that sophoraflavanone G isolated from the Greenberg et al., 2008). roots of Sophora flavescens exerts antibacterial effects against oral bacteria (Cha Ginseng root (Korean ginseng, Panax ginseng et al., 2007). C.A. Meyer) has been used for thousands of years in the traditional medical system in 2. Objective of Research oriental countries (Park, 1996; Sung, 1986). The main active components of P. ginseng are In this study, the antimicrobial activities of the triterpenoid saponins, called ginseng sophoraflavanone G isolated from Sophora saponins or ginsenosides, which have been flavescens roots and red ginseng extract shown to have a variety of biological against oral pathogens when used alone and properties including anti-inflammatory, in combination with antibiotics were assessed. antioxidant, and anticancer effects (Jung and Jin, 1996; Shibata et al., 1965; Kaneko and 3. Experimentals Nakanishi, 2004; Song, 2004). Red ginseng is made from steam heat treatment of fresh 3.1 Sample preparation ginseng roots. The red ginseng contains RGE derived from the root of P. ginseng was specific ginsenoside-Rh1, ginsenoside-Rh2, provided from KUNBO Corporation (Jinangun, 20(S)-ginsenoside Rg2, 20(S)-ginsenoside South Korea). RGE is made through a Rg3, and these are not detected or even as secondary process that concentrates the water trace amount in fresh and dried ginseng roots extract of six-year-old RG roots, and it was (Kim et al., 1991; Kim et al., 2013). These dissolved directly in 5% diluted ethanol, (1.0% specific ginsenosides of red ginseng can be as final concentration). The general used for important medicine. Ginsenosiedes composition of the product offered by KUNBO Ph ton 419 Corporation is as follows: moisture 34%, solid inhibited, were determined. Following volume 64%, ash 2.3%, total fat 0.02%, total anaerobic incubation of MICs plates, the crude saponin 70 mg/g, and ginsenosides minimum bactericidal concentrations (MBCs) (Rg1+Rb1) 20 mg/g. were determined on the basis of the lowest concentration of CT that kills 99.9% of the test The S. flavescens roots were collected in bacteria by plating out onto each appropriate October 2001, in Jinan, Jeonbuk Province, agar plate. Ampicillin (Sigma) and gentamicin Korea. The identity of the roots was confirmed (Sigma) were used as standard antibiotics in by Dr. Bong-Seop Kil at the College of Natural order to compare the sensitivity of Science, Wonkwang University. A voucher sophoraflavanone G and RGE against test specimen (JS01-3) has been deposited at the bacteria. Herbarium of the Department of Biomedicinal Chemistry and Cosmetics, at the College of 3.4 Checker-board dilution assay Technology and Natural Science of Mokwon The antibacterial effects of a combination of University. The structure of sophoraflavanone sophoraflavanone G and RGE, which G isolated from S. flavescens roots was exhibited the highest antimicrobial activity, and identified via comparisons of their spectral antibiotics were assessed by the checkerboard data (mp, MS, [ α], 1D NMR and 2D NMR) with test as previously described (Cha et al., 2007; those reported in the relevant literature (Cha et Chatterjee et al., 2009). The antimicrobial
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