J Dent Hyg Sci Vol. 13, No. 1, 2013, pp.45-52 RESEARCH ARTICLE 치아우식원인균에대한한약재추출물의항균효과 이희원ㆍ이푸른하늘 1 ㆍ권현정ㆍ한국일ㆍ한만덕 순천향대학교자연과학대학생명과학과, 1 순천향대학교자연과학대학생명공학과 Antimicrobial Activity of Extracts from Some Traditional Oriental Medicinal Plants against Dental Caries Bacteria Hee-Woen Lee, Pureunhaneul Lee 1, Hyun-Jung Kwon, Kook-Il Han and Man-Deuk Han Department of Biology, College of Natural Sciences, Soonchunhyang University, Asan 336-745, 1 Department of Biotechnology, College of Natural Sciences, Soonchunhyang University, Asan 336-745, Korea Antimicrobial activity of extract from some traditional oriental medicinal plants were evaluated for their antimicrobial activity against six oral pathogens, Streptococcus salivarius, Streptococcus oralis, Streptococcus mutans, Streptococcus sanguinis, Lactobacillus acidophilus and Lactobacillus casei, which are associated with caries disease. The antimicrobial activity was examined by determining the inhibition zone using the disc diffusion assay. In antibacterial activity test, extracts of Scutellaria baicalensis, Chrysanthemum indicum, Kochia scoparia, Hydnocarpus anthelmintica and Caesalpinia sappan showed inhibitory effects (40 mg/ml) against tested caries bacteria. Especially, the C. sappan extract showed the strongest activity on S. oralis (40 mm), L. casei (35 mm) and S. mutans (28 mm). Thus, this result suggests that C. sappan may be applicable to preventing dental caries. Key Words: Antimicrobial activity, Dental caries bacteria, Natural product, Oriental medicine 서론 미생물의발육을억제하는물질인항생물질은현재에이르기까지치료용의약품으로대량소비되고있는실정이며근래에와서항생물질의장기간투여로인한내성균으로여러가지부작용이나타나매우심각한문제점으로대두되고있다 1). 즉서양의학의경우질병발생의원인을세균과바이러스에의한것으로인식하고항생물질의개발로이를제거하거나증식을억제할수있다고생각했으나항생제의생체에대한독성과내성으로인한치료효과의감소가단점으로지적되어왔다. 이에따라미생물을이용한내성이적은항생제의개발과천연물이가지는항균활성에대한연구에많은관심이집중되어왔다 2). 천연물유래항균물질은항생 제와합성화학요법제에비해안전성이높고내성세균의출현에대한우려가적다는장점이있다 3). 그러나이러한천연항균물질의항균효과가합성된항생제에비해효과가뛰어나지않기때문에아직까지실용화되는경우는많지않은실정이다. 식물에존재하는항균물질은그대부분이 alkaloid류, flavonoid류, terpenoid류, phenolic compound류, quinone 류및 volatile oil 등의이차대사산물이거나그유도체들로알려져있다 4,5). 특히천연물에포함된성분중에서 alkaloids, polyphenols, glycosides, terpenes의 4가지가중요한작용을하는것으로알려져있으며, alkaloids는동물, 식물, 곰팡이, 박테리아를포함한다양한유기체에의해서만들어지며다른유기체에독성을보여약리효과물질로써사용되 Received: January 16, 2013, Revised: February 28, 2013, Accepted: March 1, 2013 ISSN 1598-4478 (Print) / ISSN 2233-7679 (Online) Correspondence to: Man-Deuk Han Department of Biology, College of Natural Sciences, Soonchunhyang University, 22, Soonchunhyang-ro, Sinchang-myeon, Asan 336-745, Korea Tel: +82-41-530-4702, Fax: +82-41-530-1256, E-mail: mdhan@sch.ac.kr Copyright 2013 by the Korean Society of Dental Hygiene Science This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/ by-nc/3.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
J Dent Hyg Sci Vol. 13, No. 1, 2013 어진다 6). Polyphenols은활성산소에노출되어손상되는 DNA의보호나세포구성단백질및효소를보호하는항산화능력이커서다양한질병에대한위험도를낮춘다고보고되고있다 7). Glycosides는당과다른화합물이결합된물질로식물에서여러가지역할을하며, 약리작용을가지고있는것이많다. 그리고 terpenes는항산화작용과단일산소, 수산화기, 과산화기, 초과산화기와같은자유라디칼산소에의해망가질수있는지질, 혈액그리고체액을보호하는기능을가지고있으며항균작용을한다고알려져있다 8). 치아우식증의원인은구강내에서원인세균이생성하는 glucosyltransferase에의하여당질로부터점착성의불용성글루칸이형성되어치아표면에부착하고, 글루칸에원인세균이증식함에따라국소적으로각종유기산이생성되어치아표면의법랑질을분해하는것이초기발생기전으로알려져있다 9,10). 최근천연물질에대한항우식효과에관한연구로는다양한생약추출물의항균활성 11,12) 과 manuka honey 추출물 13), Vitis amurensis 추출물 14), clove oil 추출물 15), Croton nepataefolius로부터분리된성분 16) 등의구강미생 물에대한항균활성에대한연구가보도된바있다. 본연구에서는국내에서얻을수있는 64종의한약재로부터열수추출물을분리하여치아우식증발생에관여하는 Streptococcus salivarius, Streptococcus oralis, Streptococcus mutans, Streptococcus sanguinis, Lactobacillus acidophilus, Lactobacillus casei 균주에대한항균활성을디스크확산법 (paper disc diffusion method) 으로검색하여항균제와구강청정제로서의이용가능성을알아보았다. 재료및방법 1. 실험재료본실험에사용한한약재는본초강목에서항균작용을가지고있는것으로예상되는 64종의약재를참고하여선정하였고, 실험에사용한한약재는서울경동시장에서구입하여사용하였다. 한약재명, 사용부위그리고약효는 Table 1과같다. Table 1. List of Herbal Medicine Used for Antimicrobial Activity Test Sample Korean name Scientific name Origin Healing properties SIP-1 Goihwa Sophora japonica Los Antifebrile, hypotensor SIP-2 Gilkyung Latycodon grandiflorum Radix Expectorant SIP-3 Duchung Eucommiae ulmoides Cortex Hypotensor, odynolysis SIP-4 Soksae Equisetum hiemale Herba Hypotensor, diuretic SIP-5 Ogapi Acanthopanax sessiliflorum Cortex Sthenia SIP-6 Doin Prunus persica Semen Odynolysis, antiphlogistic, antidote SIP-7 Haedongpi Kalopanax pictum Cortex Odynolysis, antimicrobial SIP-8 Cheonma Gastrodia elata Rhizoma Odynolysis, anticonvulsant SIP-9 Modkdanpi Paeonia moutan Cortex radicis Odynolysis SIP-10 Ganghwal Angekica koreana Radix Sudation, antifebrile, odynolysis SIP-11 Mogun Imperata cylindrica Rhizoma Hemostasis, diuretic SIP-12 Hwanggum Scutellaria baicalensis Radix Antifebrile, diuretic, antimicrobial SIP-13 Dansam Salvia miltiorrhiza Radix Odynolysis, angiectasis SIP-14 Baekdugu Amomum cardamomum Fructus Stomachics, antidiuresis SIP-15 Bokbunja Rubus coreanus Fructus Sthenia, antidiuresis SIP-16 Gamguk Chrysanthemum indicum Flos Antiphlogistic, diuretic, antimycotic SIP-17 Golyonpi Melia azedarrach Cortex Deworming, obstipant SIP-18 Bija Torreya nucifera Semen Deworming SIP-19 Jibuja Kochia scoparia Herba Diuretic, antifebrile, antiphlogistic SIP-20 Daepungja Hydnocarpus anthelmintica Semen Antimycotic SIP-21 Gumaengja Rosa lavigata Fructus Sthenia, cholesterol lowering SIP-22 Binlangja Areca catechu Pericarpium Antifebrile SIP-23 Ganghwang Curcuma longa Rhizoma Odynolysis, cholagogue, uterus tonica SIP-24 Baeklyom Ampelosis japonica Radix Protective liver function SIP-25 Baeksunpi Dictanmus albus Cortex Antifebrile, antimycotic SIP-26 Baekgulchae Chelidonium majus Herba Spasmolysis, inhibiting center SIP-27 Gosam Sophora flavescens Radix Indigestion, neuralgia, hepatitis SIP-28 Danggui Angelica gigas Radix Nourishing of the blood SIP-29 Guchuk Cibotum barometz Rhizoma Backache, arthritis SIP-30 Sansaja Cartaegus pinatifida Fructus Strengthen bowels 46
이희원외 : 치아우식원인균에대한한약재추출물의항균효과 Table 1. Continued Sample Korean name Scientific name Origin Healing properties SIP-31 Sanyak Discorea japonica Rhizoma Backache, diarrhea SIP-32 Sandugun Sophora subprotrata Radi Pain removal, swelling SIP-33 Sukchangpo Acorus gramineus Rhizoma Digestive fluid secretagog SIP-34 Sokdan Dipsacus asperoides Radix Blood circulation control SIP-35 Siho Bupleurum falcatum Radix Antifebrile, odynolysis, sthenia SIP-36 Sungma Cimicifuga heracleifolia Rhizoma Antifebrile, antidote SIP-37 Sini Magnolia dehunata Flos Sinus infection, chill SIP-38 GungSain Amomum xanthiodes Fructus Indigestion, diarrhea SIP-39 Yonkyo Forsythia viridisima Fructus Pus removal, diuretic SIP-40 Aeyop Artemisia asiatica Herba Hemostasis, cramps SIP-41 Omija Schizandra chinensis Fructus Stamina, sthenia, cough relax SIP-42 Osuu Evodia officinalis Fructus Headache, cold, cramps SIP-43 Indong Lonicera japonica Folium Antifebrile, enteritis, itching SIP-44 Jeonho Anthiriscus sylvestris Radix Expectorant, cough relax SIP-45 Gungang Zingerber officinalis Rhizoma Stomachache, vomiting, diarrhea SIP-46 Eumyangguk A Epimedium koreanum Herba Stamina, hypotensor, dizziness SIP-47 Eumyangguk B Epimedium koreanum Herba Stamina, hypotensor, dizziness SIP-48 Jogudong Ramulus uncariae cum uncis Lignum Hypotensor SIP-49 Jinbum Aconitum pseudolaeve nakai Radix Odynolysis, stroke SIP-50 Magamok Sorbus commixta Fructus Cough relax, arthritis SIP-51 Mahwang Ephedra sinica Herba Chill, headache, asthma SIP-52 Jinpi Fraxinus rhynchophylla hance Cortex Antiphlogistic, odynolysis SIP-53 Daehwang Rheum rhabarbarum Rhizoma Antifebrile, bile secretagog SIP-54 Chuncho Rubia akane Radix Hemostasis, pain lowering SIP-55 Chunlunja Melia azedarach L. Fructus Stomachache, didymalgia SIP-56 Chungsangja Celosia argentea L. Semen Congestion, coating on the tongue SIP-57 Chukbaeyop Thuja orientalis L. Cacumen Expectorant, hypotensor SIP-58 Donggoaja Benincasa hispida Semen Obesity, excretion SIP-59 Choyongdam Gentiana scabra Radix Indigestion, inappetence SIP-60 Chodugu Alpinia katsumadai Hayata Semen Vomiting, inappetence SIP-61 Pohwang Typha orientalis Presl Pollen Hypotensor, cholesterol lowering SIP-62 Pogongyung Taraxacum mongolicaum Herba Strengthen immunity deficiency, bile secretion SIP-63 Somok Caesalpinia sappan Lignum Blood circulation, pain removal SIP-64 Tosaja Cuscuta japonica Choisy Semen Antioxidant, anti-inflammatory SIP: sample list number. Table 2. List of Strains and Media Used for Antimicrobial Activity Test Tested strains Aerobic condition Culture media Streptococcus mutans KCTC 3065 Facultative anaerobic Brain heart infusion agar Lactobacillus casei KCTC 13086 Facultative anaerobic MRS medium Streptococcus salivarius KCTC 5512 Facultative anaerobic Nrain heart infusion agar Lactobacillus acidophilus KCTC 3140 Facultative anaerobic MRS medium Streptococcus sanguinis KCTC 3284 Facultative anaerobic Brain heart infusion agar Streptococcus oralis KCTC 13048 Facultative anaerobic Trypticase soy yeast extract medium KCTC: Korean Collection for Type Cultures. Brain heart infusion agar (Difco Lab., Detroit, MI, USA), MRS medium (Difco Lab.), trypticase soy yeast extract medium (Difco Lab.). 2. 사용균주및배지본실험에사용한균주는한국생물자원센터 (Korean Collection for Type Cultures) 에서분양받아사용하였다. 각균주에대한생육배지는 S. mutans, S. salivarius, S. sanguinis 는 brain heart infusion agar (Difco Lab., Detroit, MI, USA) 를사용하였고, L. casei, L. acidophilus는 MRS medium (Difco Lab.) 을사용하였으며, S. oralis는 trypticase soy yeast extract medium (Difco Lab.) 을사용하였다. 47
J Dent Hyg Sci Vol. 13, No. 1, 2013 사용할균주는혐기자에 Anaerobe Container System (BD GasPaK EZ, Becton, Dickinson and Co., Sparks, MD, USA) 과함께넣어 37 o C에서 24시간 3번계대배양하였다. 배양한모든균주는분광광도계 (Libra S12, Biochrom Ltd., Cambridge, UK) 를이용하여흡광도 (optical density) 값을 0.5로맞춘후사용하였다 (Table 2). 3. 항균력측정을위한한약재추출물의조제한약재의추출물은앞서언급한한약재분쇄물을열탕추출하여항균활성검색을위한시험재료로사용하였다. 열수추출법은천연물로부터극성물질뿐만아니라많은양의비극성물질도수용액중에용출되므로 1차생리활성검색을위한시료조제법으로활용된다. 따라서한약재의항균력을알아보기위한추출물조제는각각의한약재 10 g 당증류수 300 ml을혼합하여 100 o C water bath에서약 3시간중탕한다음 Whatman No. 2 (Whatman International Ltd, Maidstone, UK) 여과지를사용하여 1차여과과정을거친후 pore size 0.45 μm syringe용멸균필터를사용하여무균적으로한약재열수추출액을조제하였다. 위과정을거쳐얻어진시료를감압농축시킨후동결건조하여 4 o C에서보관하였다 (Fig. 1). 4. 치아우식균에대한한약재의항균효과측정한약재의생리활성검색은디스크확산법으로측정하였다. 즉각각의균주들을 20 ml 생육배지에서 18시간정도배양하여병원성균배양액을 620 nm에서흡광도 (optical density) 값 0.5가되게분광광도계로조절후멸균된면봉으 Fig. 1. Extraction and fractionation of medicinal herbs. 로회전하면서배지전면에균을골고루도말하였다. 각각의시료는멸균된 Eppendorf tube에 40 mg/ml이되도록멸균된증류수로녹여시료를만들었다. 시료는멸균된 paper disc; 8 mm (Advantec, Toyo Roshi Kaisha Ltd., Tokyo, Japan) 에 100 μg씩흡수시킨후평판배지에 paper disc를밀착시키고, 37 o C incubator에서 24시간배양한다음 paper disc 주변의생육저해환 (inhibitory zone, clear zone) 의직경을측정하여항균력을비교하였다. 결과 1. 한약재추출물의 Streptococcus 속 ( 연쇄상구균속 ) 균주에대한항균성검색한약재 64종류로부터얻은열수추출물의 4종의 Streptococcus 속우식유발균주에대한항균활성은디스크확산법을이용하여확인한결과, 다음과같다. S. mutans에항균활성을나타낸한약재추출물의생육저해환의직경은황금 (SIP-12, Scutellaria baicalensis) 에서 8 mm, 소목 (SIP-63, Caesalpinia sappan) 에서 28 mm로나타났다. 초기평활면우식증에영향을주는 S. sanguinis 또한 64종의한약재추출물가운데황금 (SIP-12, S. baicalensis) 과소목 (SIP-63, C. sappan) 만이항균활성을나타냈으며, 황금 (SIP-12, S. baicalensis) 에서 10 mm, 소목 (SIP-63, C. sappan) 에서 17 mm의생육저해환의직경을측정하였다. S. oralis에대한한약재추출물의항균활성결과, 64종가운데 16개의한약재가항균활성을보였다. 항균활성이가장우수한한약재는소목 (SIP-63, C. sappan); 40 mm이었고, 다음으로감국 (SIP-16, Chrysanthemum indicum); 27 mm, 황금 (SIP-12, S. baicalensis); 19 mm, 지부자 (SIP-19, Kochia scoparia); 17 mm, 대풍자 (SIP-20, Hydnocarpus anthelmintica); 17 mm 순으로나타났다. 이밖에도괴화 (SIP-1, Sophora japonica); 14 mm, 오가피 (SIP-5, Acanthopanax sessiliflorum); 9 mm, 목단피 (SIP-9, Paeonia moutan) 14 mm, 단삼 (SIP-13, Salvia miltiorrhiza); 10 mm, 비자 (SIP-18, Torreya nucifera); 10 mm, 고삼 (SIP-27, Sophora flavescens); 10 mm, 시호 (SIP-35, Bupleurum falcatum); 11 mm, 인동 (SIP-43, Lonicera japonica); 10 mm, 음양곽A (SIP-46, Epimedium koreanum); 13 mm, 음양곽B (SIP-47, E. koreanum); 11 mm, 대황 (SIP-53, Rheum rhabarbarum); 13 mm이항균활성을보였다. S. salivarius 는천마 (SIP-8, Gastrodia elata); 9 mm, 소목 (SIP-63, C. sappan); 20 mm의생육저해환의직경을측정하였다 (Table 3, Fig. 2). 48
이희원외 : 치아우식원인균에대한한약재추출물의항균효과 Table 3. Antimicrobial Activity of Extracts from the Oriental Medicinal Plants against Caries Bacteria by Disc Diffusion Assay a SIP Zone of inhibition b (mm) (40 mg/ml) 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 Strains 1 - - - - - - - + - - - - - - - 2 ++ - - - + - - - ++ - - +++ + - - 3 - - - - - - - - - - - + - - - 4 - - - - - - - - - - - + - - - 5 - - - - - - - - - - - - - - - 6 - - - - - - - - - - - + - - - SIP 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 Strains 1 - - - - - - - - - - - - - - - - 2 ++++ - + +++ +++ - - - - - - + - - - - 3 - - - - - - - - - - - - - - - - 4 - - - - - - - - - - - - - - - - 5 - - - - - - - - - - - - - - - - 6 - - - - - - - - - - - - - - - - SIP 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 Strains 1 - - - - - - - - - - - - - - - - 2 - - - ++ - + - ++ ++ + ++ + - - ++ ++ 3 - - - - - - - - - - - - - - - - 4 - - - - - - - - - - - - - - - - 5 - - - - - - - - - - - - - - - - 6 - - - - - - - - - - - - - - - - SIP 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 64 Strains 1 - - - - - - - - - - - - - - - +++ - 2 - - - - - ++ - - - - - - - - - ++++ - 3 - - - - - - - - - - - - - - - ++++ - 4 - - - - - - - - - - - - - - - +++ - 5 - - - - - - - - - - - - - - - ++ - 6 - - - - - - - - - - - - - - - ++++ - a The antimicrobial activity was represented as followed: -, no inhibitory zone; +, <11 mm; ++, <15 mm; +++, 20 mm; ++++, >20 mm, b Diameter of clear zone including disc diameter of 8.0 mm. SIP: sample list number, strains 1: Streptococcus salivarius, 2: Streptococcus oralis, 3: Streptococcus mutans, 4: Streptococcus sanguinis, 5: Lactobacillus acidophilus, 6: Lactobacillus casei. 2. 한약재추출물의 Lactobacillus 속 ( 유산간균속 ) 균주에대한항균성검색한약재 64종류로부터얻은열수추출물의 2종의 Lactobacillus 속우식유발균주에대한항균활성결과는다음과같다. 치면열구우식증에연관성이높은 L. casei에대한항균활성은 64종의한약재추출물가운데황금과소목에서뚜렷한생육억제활성을나타냈다. L. casei는황금 (SIP-12, S. baicalensis); 9 mm, 소목 (SIP-63, C. sappan); 35 mm의생육저해환의직경을측정하였다. 소와열구및상아질우식에관여하는 L. acidophilus는 64종한약재추출물가운데 소목 (SIP-63, C. sappan) 만이항균성을보였다 (Table 3, Fig. 2). 3. 소목과황금의우식유발균주에대한항균활성항균활성시험검색결과, 소목 (SIP-63, C. sappan) 과황금 (SIP-12, S. baicalensis) 은대부분의 Streptococcus 속 ( 연쇄상구균속 ) 균주와 Lactobacillus 속 ( 유산간균속 ) 균주에대해활성을나타내는것으로나타났다. 특히소목 (SIP-63, C. sappan) 의경우 S. oralis와 L. casei에서 40 mm와 35 mm의생육저해환의직경을측정하였고, 황금 (SIP-12, S. 49
J Dent Hyg Sci Vol. 13, No. 1, 2013 Fig. 2. Antimicrobial activity of medical herb against Streptococcus salivarius (A), Streptococcus mutans (B), Streptococcus sanguinis (C), Streptococcus oralis (D G), and Lactobacillus casei (H) by the disc diffusion assay. Table 4. Antimicrobial Activity of SIP-63 (Caesalpinia sappan) and SIP-12 (Scutellaria baicalensis) against caries Bacteria by Disc Diffusion Assay Tested strain SIP-63 SIP-12 Zone of inhibition a (mm) Streptococcus salivarius 20 - Streptococcus oralis 40 19 Streptococcus mutans 28 8 Streptococcus sanguinis 17 10 Lactobacillus acidophilus 13 - Lactobacillus casei 35 9 a Diameter of clear zone including disc diameter of 8.0 mm. -, no inhibitory zone. baicalensis) 의경우 S. oralis에서 19 mm의생육저해환을측정하였다 (Table 4). 고찰 치아우식증과치주질환은주요구강내질환으로, 치아우식증원인균으로 Streptococcus species (spp.) 및 Lactobacillus spp. 에속하는일부의종들이보고되고있으며, 그중 S. mutans가가장밀접한관련이있는것으로알려져있다 17,18). 이전의연구에의하면 Lactobacillus 종들은초기치아우식증보다는진행된치아우식증병소와더연관성이높고, 치근면치아우식증을포함한모든치면의치아우식증에는 S. mutans가주요한원인세균으로알려져있다 19,20). 치아우식증과치주병은항균및항우식성분이포함된구강위생용품을적절히이용한다면예방이가능하다. 천연식물추출물이포함된구강위생용품은독성이있고, 치아에착색을유발하는 triclosan, cetylpyridinium chloride, chlorhexidine 그리고 amine fluorides와같은항균제제가포함된구강위생용품에비해독성이낮기때문에관심이높아지고있다 21,22). 약재내성의문제와구강위생용품에의적용을위한새로운항균물질로서 tannins, terpenoids, alkaloids, 및 flavonoids 와같은식물추출물에의해생성되는많은대사물질이개발되고있는실정이다 23-25). 본연구는 6종의치아우식유발균주에대한 64종의한약재추출물의균별항균력을조사하기위해각각의추출물을 40 mg/ml로희석하여디스크확산법으로 clear zone을확인한결과, 소목추출물은모든균주에대해항균활성을나타내었고특히 S. oralis와 L. casei의경우다른균주와비교해볼때 40 mm, 35 mm로가장큰 clear zone을형성하였다. 권등 26-28) 의연구는소목추출물중에틸아세테이트 (EtOAc) 분획에존재하는 brazilin이병원성균주에대한항균력을가지고있으며, 소목의에틸아세테이트분획물이 S. mutans의항균활성및부착억제효과를가진다고보고하였다. 또한소목에서분리된 brazilin이 S. mutans의생장을억제하는효과를가지는것으로나타나우식원성세균의억제물질로서의활용가능성을제시한바있다. 황금추출물은 S. 50
이희원외 : 치아우식원인균에대한한약재추출물의항균효과 salivarius와 L. acidophilus를제외한나머지균주에서모두항균활성을나타냈으며, S. oralis에서 19 mm로가장큰활성을보여주었다. 백등 29) 은황금추출물 50 mg/ml을투여한배지에서 S. mutans에대한생장을지연시켰으며, 최소억제농도인 125 mg/ml에서 hydroxyapatite (HA) 에 S. mutans의부착억제효과를가지는것으로나타났다. 이는본연구와유사한농도에서항균활성을가지는것으로판단되며, 최소억제농도에대한추가적인실험이필요할것으로사료된다. 가장많은종류의천연물에생육저해활성을보인 S. oralis는황금 (19 mm), 감국 (27 mm), 지부자 (17 mm), 대풍자 (17 mm), 소목 (40 mm) 에서높은항균활성을나타냈다. 따라서다른균주와비교하였을때천연물에대한민감도높은것으로보인다. 본연구의제한점으로는실험방법으로서 disc diffusion assay의경우배지내습도등으로인하여 disc에주입한추출물의확산이불규칙할수있기때문에다양한방법으로항균물질의항균력을측정할필요가있는것으로생각된다. 따라서다양하고새로운유용식물자원을발굴하여체계적인생리활성검색과활성물질의분리및구조규명작업이앞으로지속적으로이행해나가야할것이다. 본연구를통해소목과황금추출물은구강내치아우식유발균주의생육억제활성이우수하여차후다양한방법으로천연항균물질로서의구강내제재개발의사용가능성에대한연구가계속되어야할것으로생각된다. 요약 본연구에서는, 구강내세균에대한항세균제로서의이용가치를확인하고, 치아우식예방및치료제로활용될수있는가능성이있는물질을탐색하고자 64종의한약재추출물을사용하여치아우식증의원인이되는 6개균주를대상으로항균활성을검색하였다. 1. 구강내상주균인 S. oralis, 치아우식증에가장큰영향을미치는 S. mutans, 치면열구우식증에연관성이높은 L. casei와초기평활면우식증에영향을주는 S. sanguinis에모두항균활성을보인한약재추출물은 64종가운데황금과소목이었다. 2. 치아우식증원인균중하나인 S. oralis에대하여한약재추출물의항균활성을알아본결과 64종가운데 22개의한약재가항균활성을보였다. 항균활성이가장우수한한약재는소목 (40 mm)>감국 (27 mm)>황금 (19 mm)>지부자 (17 mm), 대풍자 (17 mm) 순이었다. 그외에괴화 (14 mm), 오가피 (9 mm), 목단피 (14 mm), 단삼 (10 mm), 비자 (10 mm), 고삼 (10 mm), 시호 (11 mm), 인동 (10 mm), 음양곽 A (13 mm), 음양곽 B (11 mm), 대황 (13 mm) 이항균활성을보였다. 3. 소와열구및상아질우식에관여하는 L. acidophilus는 64종한약재추출물가운데소목만이항균성을보였다. 치아우식증원인세균인 S. salivarius는천마 (9 mm), 소목 (20 mm) 의생육저해환의직경을측정할수있었다. 4. 결론적으로, 치아우식증의원인이되는 6종균주에대해가장우수한활성을보인한약재는소목으로최대 40 mm의생육저해환이측정되었다. 이상의결과를볼때, 한약재추출물은구강내우식유발균주의생육을억제하는효과가있어, 향후구강내우식원성세균의치료물질로활용을제시한다. 감사의글 본연구는순천향대학교학술연구비의일부지원으로수행하였음. 참고문헌 1. Stuart BL: The antibiotic paradox: How miracle drugs are destroying the miracle. Plenum publishing Co., New York, pp.114-129, 1992. 2. Brumfitt W, Hamilton-Miller J: Methicillin-resistant Staphylococcus aureus. N Engl J Med 320: 1188-1193, 1989. 3. Cho JY, Choi I, Hwang EK: Antimicrobial activity of extracts from medicinal herbs against Escherichia coli. Kor J Vet Res 43: 625-631, 2003. 4. Mitscher LA, Park YH, Clark D: Antimicrobial agents from higher plants, antimicrobial isoflavonoids and related substances from Glycyrrhiza glabra L. var Typica. J Nat Prod 43: 259-269, 1980. 5. Lee BW, Shin DH: Screening of natural antimicrobial plant extract on food spoilage microorganism. Kor J Food Sci Technol 23: 200-204, 1991. 6. Lee MK, Sung HG, Hwang WG, Kim IH: Alkaroids from the tuber of Aconitum chiisanense. Korean J Pharmaceutial 41: 161-173, 1997. 7. Bub A, Watzl B, Blockhaus M, et al.: Fruit juice consumption modulates antioxidative status, immune status and DNA damage. J Nutr Biochem 14: 90-98, 2003. 8. Osbourn AE, Lanzotti V: Plant-derived natural products: 51
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