Antimicrobial Effect of Puerariae thunbergiana Extracts against Oral Microorganism

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(1)Journal of Dental Hygiene Science Vol. 4, No. 1, pp. 45~ 48 (2004). W ºÂb~ \;bö Î. *K ÁB?\ Áªêj Á\ ÁWßz

(2). †. ö% ~* 9 ~* ö%L ~*^ö £¶öB*. 1. 2. Antimicrobial Effect of Puerariae thunbergiana Extracts against Oral Microorganism †. Hyun-Ok Lee , Chang-Hee Kim1, Jin-A Lim2, Mi-Hee Lee2 and Seung-Hwa Baek2 Department of Dental Hygiene, Wonkwang Health Science College, Iksan 570-750, Korea 1 Department of Dental Hygiene, Shinsung College, Dang Jin 343-861, Korea 2 Department of Herbal Resources, Professional Graduate School of Oriental Medicine and Institute of Basic Natural Sciences, Wonkwang University, Iksan 570-749, Korea. " "#! #!#. In the current research for natural products with antimicrobial effects, various extracts of Puerariae thunbergiana and isoflavones against microorganisms were evaluated in terms of the minimum inhibitory concentrations (MIC). In general, Candida albicans was stronger antimicrobial activity than the other microorganisms such as Streptococcus mutans, Staphylococcus epidermidis, and Staphylococcus aureus. The maximum activity was exhibited by methanol extract of the leaves of Puerariae thunbergiana Beth. against Candida albicans(MIC, 400 µg/mL). These results suggest that methanol extract of Puerariae thunbergiana has a potential antimicrobial activity. ,FZ XP XPSE SETT Puerariae thunbergiana, Antimicrobial effects, Minimum inhibitory concentration(MIC). *. b~ Î"ö & º j ç . º ~ òÒ, *Vf ´j FVÏ (n-hexane, chloroform, ethyl acetate, methanol) ³ºÂ~ áf ºÂb" ÒW bî ;bö &~ Î"¢ áîVö ~º : .. . ."(Puerariae Radix)f ÖÒ¾¢¢ j~ 7 5 ¢ öB ¶~º v"b(Leguminosae), j ¿#W # ¾Z (Puerariae thunbergiana Beth.)~ òÒ ¦ªj © . ."öº 10~15%ö ~º *ª" 9 «~~ saponin, .& {Ë, .~ ;~, ·Ï j ¾æ Úº flavonoids, Ò isoflavonoid daidzin" "G~ Z Ë¾ ãNj jzÊº êã·Ïj ¾æÚº daidzein, & Ë ô FB puerarin b W>Ú ® . ~öB ~ ."f ÓïW BBB ç, 7³ ., 7V~ vÛ, ., î, Û, B, Z ö Ï>, $ JÒ¢ nº ² ~º çÎ& ®b, Î £ëj B rJ^ ®. . O~öBº .{, &çÿ ãzÃ, 7Ã, W ö Ï~ ®b , .{;~·Ï , æO~ ÖzÛB· Ï, "·Ï, ë·Ï, ?B~ Î", Öz·Ï 5 * ·Ï ~ Wj &V B : ®b¾. Òò5O» 1.. 2). þÒò. þö ÒÏB (Puerariae thunbergiana Beth.)f 2002 j 10úö ÏÎ ¢Ö W &ÒÖöB j÷êö r ~ ¦;¢ ¦; ê, Blender ªê~ ÒÏ~& .. þö ÒÏB bÚº ö7&v ~*^&ö £¶ö BB"ö &>Ú ®º Òò¢ ÒÏ~& .. 1). 3). þO» ºÂO» ö ÒÏB (òÒ, *V, ´)j ^.~ Blender ªê~ ÒÏ~& . ~ ¦*ê n-hexaneb çNöB 24*ÿn êû~B 3² > ºÂ ê, ÖºÂj 0.4 µm "æ "~ 35 CöB 6{³»~ ÖºÂbj 2.. 4-6). 1). †. Corresponding author Tel: 82-63-840-1265 Fax: 82-63-840-1269 E-mail: [email protected]. o. .

(3) . Antimicrobial Effect of Puerariae thunbergiana Extracts against Oral Microorganism. áî . *f ?f O»b ª, ö j^rÞ, z êR ºÂ~, ª ºÂb, ö j^rÞ ºÂb, zêR ºÂbj áî . ."û" ßî."ûf 80 C~85 Cö B 4*ÿn ~~ºÂ~ b ºÂb" 50% öêR ºÂb j áî . 2) VV 5 £ Rotary vaccum evaporator(EYELA : N-N Series, Swizerland), freezing dryer(-50 C ~, vaccum 5 microns Hg ~ : F. D8510, Ilshin Lb Co.), CO incubator (NUAIRE), o. o. o. 2. ELISA reader(Molecular devices, spectra MAX 340, USA), Microscope(Olyimpus, CK2, USA). n -hexane, chloroform, ethyl acetate, methanol(DAEJUNG) , 3 . Daidzein quercetin Aldrich (U.S.A) . 3) Streptococcus mutans(S. mutans, JC-2), Staphylococcus epidermidis(S. epidermidis, ATCC 12228), Staphylococcus aureus (S. aureu, ATCC 29213) Candida albicans (C. albicans, KCTC 1940) . S. mutans, S. epidermidis S. aureus Brain heart infusion (BHI, Difco) , C. albicans Sabouroud( 1L Bacto-peptone 10 g, Bacto - dextrose40g) . 4) (minimum inhib7-9) itory concentration, MIC) . , 96-microwell plate(Nunclon, Denmark) 3,200 µg/mL 100 µg/ mL 2 . , 37oC 18 105 cells/ml 37oC 24 , ELISA(Spectra Max 250, Molecula Deveces, USA) 630 nm , MIC , MIC . 5) 4oC , 10% DMSO .. f ÒÃ~~ ÒÏ~&b bf N Ã~>¢ ÒÏ~&. " f Ò öB «~ ÒÏ~&. ÒÏ" 5 Væ. þö ÒÏB "º f ¢ ÒÏ~&. ^V·ö ÒÏB Væº f º ¢ ÒÏ~& º Ã~> ö ¢ ÒÏ~&. K G; Î"¢ 2k~V *~ ²ÛB³ê ¢ G;~& ÚVæ \C» j Ï~&b ö' ºÂb~ ³ê¢ öB &³ê ræ VO ³'b \C~& ' "º ¢ R î¢ ÚVæö 7«~ V·VöB *ÿn V· "¢ 7«~ V·VöB * V· ê ö ~ W>2Ë öB W7ê¢ G;~ Væ~ çê¢ {~& B> V·~ W7ê 8" ?f Ö"¢ áf ©j Ö;~&b ~ >~& Ôf ©j Î"& ¸f ©b 6;~&. ò~ ¾Ò B òº ¯ ïËö &Ë~& ÒÏç*ö ö \C~ þö ÒÏ~&. ö & ²ÛB³êº 3,200 µg/mL çb W ¾æ¾æ p~ . ¾ C. albicansº ª ºÂböB ²ÛB³ê& 800 µg/mLb êW &V >î . ö j^rÞ ºÂb" zêR ºÂbö &~ S. mutans, S. epidermidisf S. aureus & ²ÛB³êº 3,200 µg/mLb W ¾æÒ . ê C. albicansº Ö ºÂbj B~º Î ºÂböB ²ÛB³ê& ¸f 800 µg/mLb êW &V>î . òÒ ºÂ º Âbf ÎÎ; ê C. albicansö ê Î"j F ÒWbî >Ú ®bÒ¢ 'B (Table 1). S. aureus. 5BCMF Minimal inhibitory concentrations(MIC) of the root. of Puerariae thunbergiana Beth Tested material HXRP CFRP EARP MTRP *. 13). >3,200 >3,200 3,200 3,200. S. aureus. C. albicans. >3,200 >3,200 3,200 3,200. >3,200 800 800 800. >3,200 >3,200 3,200 3,200. Each extract was examined in tripliacte experiments. Plant extracts: HXRP; n-Hexane extract of the root of Puerariae thunbergiana Beth. CFRP; Chloroform extract of the root of Puerariae thunbergiana Beth. EARP; Ethyl acetate extract of the root of Puerariae thunbergiana Beth. MTRP; Methanol extract of the root of Puerariae thunbergiana Beth.. 5BCMF Minimal inhibitory concentrations(MIC) of the trunk. of Puerariae thunbergiana Beth Tested material HXTP CFTP EATP MTTP. 10-12). 13,14). S. mutans S. epidermidis. ~ *V¢ b" FVÏ ºÂ~ áf ºÂböB Î"¢ G; Ö", ÖºÂb, ª ºÂb" ö j^rÞºÂbö &~ S. mutans, S. epidermidisf S. aureus ~ ²ÛB³êº 3,200 µg/mL çb W ¾æ¾æ p~ . ¾ C. albicansº ö j^rÞ ºÂ böB ²ÛB³ê& 1,600 µg/mLb W &V>î. . W ¸f zêRºÂb~ ãÖ S. mutansf C. albicans º ²ÛB³ê& 800 µg/mLî, S. epidermidisf S. aureus~ ²ÛB³êº 3,200 µg/mL ¾æÒ . V¢B WÏ ºÂ ºÂböB 5 êbî F>îb Ò¢ 'B (Table 2).. Ö5V. ~ òÒ, *Vf ´j FVÏ ³ºÂ~ áf º Âb Ï~ ;b 7 ~jÖö S. mutans , ; vf b¦ö ç"B z³W"Ã~ ö S. epidermidisf S. aureus , ;ö ç"~B K & ~B Ò²ö² V²6"j ¾æÚº ö C. albicans ö &~ ê Î"¢ 2k~& . ~ òÒ¢ b" FVÏ ¢ ÒÏ~ W Ôf Ï ¦V W ¸f Ï çNöB ³ºÂ~ áf ºÂb öB Î"¢ G; Ö", W Ôf Ö" ª Ï ºÂ ºÂbö &~ S. mutans, S. epidermidisf. MIC50 (µg/mL)*. *. MIC50 (µg/mL)* S. mutans >3,200 >3,200 >3,200 800. S. epidermidis >3,200 >3,200 >3,200 3,200. S. aureus >3,200 >3,200 >3,200 3,200. C. albicans >3,200 >3,200 1,600 800. Each extract was examined in tripliacte experiments. Plant extracts: HXTP; n-Hexane extract of the trunk of Puerariae thunbergiana Beth. CFTP; Chloroform extract of the trunk of Puerariae thunbergiana Beth. EATP; Ethyl acetate extract of the trunk of Puerariae thunbergiana Beth. MTTP; Methanol extract of the trunk of Puerariae thunbergiana Beth.. ~ ´j b" FVÏ ºÂ~ áf ºÂböB Î"j G; Ö", ÖºÂb, ª ºÂb" ö j^rÞºÂbö & S. mutans, S. epidermidisf S..

(4) . Journal of Dental Hygiene Science Vol. 4, No. 1, pp. 45~48 (2004). ~ ²ÛB³êº 3,200 µg/mL çb W ¾æ¾æ p~ . ê C. albicansº Ö ºÂböBò ²ÛB³ê& 3,200 µg/mLb êW &V>î . ~ ´ zêR ºÂb~ ãÖ, C. albicans~ ²ÛB³êº 400 µg/mLî, S. mutans~ ²ÛB³êº 1,600 µg/mL, S. epidermidisf S. aureus~ ²ÛB³êº 3,200 µg/mLb ¾æÒ . ~ òÒ, *V 5 ´j b" FVÏ ºÂ ºÂb~ 5 êK~ ²ÛB³ê 7ö ´~ z êRºÂböB ÎÎ; ê C. albicanöB &Ë ¸f ²ÛB³ê 400 µg/mL& &V>î (Table 3). aureus. 5BCMF Minimal inhibitory concentrations(MIC) of the leaves. of Puerariae thunbergiana Beth. *. MIC50 (µg/mL)*. Tested material. S. mutans. HXTP CFTP EATP MTTP. >3,200 >3,200 >3,200 1,600. S. epidermidis >3,200 >3,200 >3,200 3,200. S. aureus C. albicans >3,200 >3,200 >3,200 3,200. 3,200 >3,200 >3,200 400. Each extract was examined in tripliacte experiments. Plant extracts: HXTP; n-Hexane extract of the leaves of Puerariae thunbergiana Beth. CFLP; Chloroform extract of the leaves of Puerariae thunbergiana Beth. EALP; Ethyl acetate extract of the leaves of Puerariae thunbergiana Beth. MTLP; Methanol extract of the leaves of Puerariae thunbergiana Beth.. ."j Vb ¾OB ."û" ßî."ûj b" 50% öêR ºÂ ºÂbö & Î"¢ G; Ö", ¢> 'b ;~ Î "ö &~ ."û ßî."û. 5 êK jv' ¸² ¾æÒ . ¯ ~ òÒ, * V 5 ´öB ºÂ ºÂb~ êÎ"& &VB ©" ? ÎÎ; ê C. albicanöB jv' ¸f êÎ"& ¾æÒ . f ?f þÖ"º ."û" ßî."ûö ¾O B £Ò& ." ~ £Òö VB êWbîö ~ ©b '>, b ."û" ßî."ûö Fê £Òö & ¦Æ& Ú^¢ ~Ò¢ 'B (Table 4). 5BCMF Minimal inhibitory concentrations(MIC) of galgeuntang. and seungma-galgeuntang Tested material WTGT ETGT WTSM ETSM ETRP *. MIC50 (µg/mL)* S. mutans. S. epidermidis. 1,600 1,600 3,200 3,200 3,200. 3,200 1,600 3,200 3,200 3,200. S. aureus C. albicans 3,200 1,600 1,600 3,200 3,200. 800 800 800 1,600 800. Each extract was examined in tripliacte experiments. Plant extracts: WTGT; Water extract of galgeuntang. ETGT; 50% Aqueous ethanol extract of galgeuntang. WTSM; Water extract of seungmagalgeuntang. ETSM; 50% Aqueous ethanol extract of seungmagalgeuntang. ETRP; Ethanol extract of the root of Puerariae thunbergiana Beth.. ."öB ªÒB isoflavonoid puerarin, daidzinö &~ S. mutans, S. epidermidisf S. aureusö & ²ÛB ³êº 3,200 µg/mLb £ W ¾æÒ . º â~ ª ººÂb, zêR ºÂb" öj^rÞ 18). 15). ºÂbö &~ S. mutans, S. epidermidisf S. aureusö & ²ÛB³ê& 25 µg/mLî ~&, f VzN®V~ ª ºÂböB ²ÛB³ê& 100 µg/mLî ~&b, f WÂ~ öêR ºÂb, ª ºÂb" öj^rÞ ºÂb S. mutansö &~ ²ÛB³ê& 500 µg/mLî : ® . f?f Ö"¢ jv~, ºÂb â~ ºÂb ~ Î"& z Ö> ©b 'B . ~ WWª daidzein" quercetin~ ²ÛB³êº 3,200 µg/mL çb W ¾æ¾æ p~b¾, ê C. albicansö & ²ÛB³êº 800 µg/mL-3,200 µg/mLb ¾æÒ (Table 5). Ö"'b ~ òÒ, *Vf ´~ ³ºÂböB êÎ "º ê C. albicansö &~ S. mutans, S. epidermidis f S. aureus ¸f W &V>îb, ~ WWª b B isoflavonoid W ¸² G;>î . f ?f þÖ"º ö êWj ¾æÚº ÒWbî puerarin, daidzin, daidzein, quercetin º Wbî F>Ú ¾æ¾º ©b 'B . Ëê ºÂbö & 5 êbîj ªÒ 5 ªC~, Wbîö & ;b~ ÛBÎ"¢ ~¶ . 16). 17). 5BCMF Minimal inhibitory concentrations (MIC) of pure. compounds MIC50 (µg/mL)*. Tested material. S. mutans. S. epidermidis. Puerarin18,19) Daidzin18,19) Daidzein Quercetin. 3,200 3,200 >3,200 >3,200. 3,200 3,200 >3,200 >3,200. *. S. aureus C. albicans 3,200 3,200 >3,200 >3,200. 3,200 800 >3,200 >3,200. Each extract was examined in tripliacte experiments.. Ö. . öBº ~ òÒ, *Vf ´j FVÏ ³ºÂ ~ áf ºÂb" WWªbîj Ï~ ;bö & 5 êÎ"¢ ¦ï~ r" ?f þÖ"¢ áî . ~ *Vf ´~ zêRºÂböB S. mutans ö & Î"º ²ÛB³ê& 800 µg/mLf 1,600 µg/mLb ¾æ Òb¾, ~ òÒöB~ ²ÛB³êº 3,200 µg/mL ¾æ Ò . ~ òÒ 50% öêR ºÂb" ßî."û bºÂbö B S. epidermidis, S. aureusö & ²ÛB³êº 1,600 µg/mLb W ¾æÒb, ´~ zêRºÂbö & C. albicans~ ²ÛB³êº 400 µg/mLb &Ë ¸ f êÎ"& &V>î . Ö"'b ~ òÒ, *Vf ´~ ³ºÂböB Î"º S. mutans, S. epidermidisf S. aureus ê C. albicansö &~ ¸f êW &V>îb, $ ~ WWªb B isoflavonoid êW ¸² G;>î . f ?f êKf ö êWj ¾æÚº ÒWbî isoflavonoid ¾ flavonoid º Wbî F>Ú ¾æ¾º ©b 'B ..

(5) . Antimicrobial Effect of Puerariae thunbergiana Extracts against Oral Microorganism. 6Ò~. ö ÒÏB ."û" ßî."ûj B~ " ã Î cö JJ~öö² 6ÒÒ, 2003jê ö7& jö ~ Úrb, ö 6ÒÖ . ^^ò. Ëç^ £¶öb ^Â$. 1. 4 : , , 1996. 2. Huh J: Donguibogam, Namsandang, Vol. 3, 1984. 3. Fan LL, Dennis DO and Wand WJP: Pharmacologic studies on Radix of Puerariae. Effect of Puerariae on reginal myocardial blood-flow and cardic hemodynamics in dogs with acute myocardial ischemia. Chin Med J 98: 821-832, 1985. 4. Oh MJ, Lee KS, Son HY and Kim SY: Antioxidative components of Pueraria root, Korean J Food Sci Technol 22: 793-798, 1995. 5. Han SH, Kin JB, Min SG and Lee CH: The effects of Puerariae Radix catechins administration on liver function in carbon tetrachloride-treated rats. Korean J Soc Food Nutr 24: 713-720, 1995. 6. Lee JS, Lee KH and Jeong JH: Effects of extracts of Puerariae Radix on lipid metabolism in rats fed high fat diet. J Korean Soc Food Sci Nutr 28(1): 218-224, 1999. 7. Cho H, Weon RS, Yang EY and Kim JS: Antimicrobial effect of the extract of Sophora flavescens Ait (I). J Pharm Soc Korea 43: 419-422, 1999.. 8. Kim KY, Chung DO and Chung HJ: Chemical composition and antimicrobial activities of Houttuynia cordata Thunb. Korean J Food Sci Technol 29(3): 400-406, 1997. 9. Chung BS, Kim HS and Kim SK: Susceptibility tests of Candida species isolated from vagina. J Korean Soc Microbiol 24(5): 523526, 1989. 10. Hamada S, Koga T and Ooshima T: Virulence factors of Streptococcus mutans and dental caries prevention. J Dent Res 63: 407-411, 1984. 11. De Stoppelaar JD, Konig KG, Plasschaert AJM, van der Hoeven JJ: Derease in cariogenicity of Streptococcus mutans. Arch Oral 53: 1355-1360, 1974. 12. Loesche WJ: Role of Streptococcus mutans in human dental decay. Microbial Rev 50: 353-380, 1986. 13. , : , , 183-184, 1998. 14. , , : , , 382-383, 1990. 15. : . , 39-40, 2001. 16. 7 : . 32(1): 55-60, 2001. 17. 8 : . 16(2): 348-352, 2002. 18. Lee SJ, Baek HJ, Lee CH and Kim HP: Antiinflammatory activity of isoflavonoids from Pueraria Radix and biochanin A derivatives. Arch Pharm Res 17: 31-35, 1994. 19. Oh MJ, Lee KS, Son HY and Kim SY: Antioxidative components of Pueraria root. Kor J Food Sci Technol 22: 793-798, 1990. (Received June 9, 2004; Accepted June 16, 2004). B'² ò{ ;b ^Ò "ã\ B~ã `"8 &b ö *K £ÏbR¦8 ºÂ Âbî~ Î"f zW ö7&v ;Ò* ¢^ "v VzN®8 ºÂb~ ^

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