DBPIA-NURIMEDIA

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1 KOREAN J. FOOD SCI. TECHNOL. Vol. 43, No. 6, pp. 760~765 (2011) The Korean Society of Food Science and Technology Paraquat 에의해유발된산화적스트레스에의한흑마늘추출물의항산화효과 노병규 이정규 원용덕 1 박현진 이성준 * 고려대학교생명과학대학식품공학부, 1 의성흑마늘영농조합 The Antioxidative Effect of Black Garlic Extract on Paraquat-induced Oxidative Stress in ICR Mice Byung-Kyu Noh, Jung Kyu Lee, Yong-Duk Won 1, Hyun-Jin Park, and Sung-Joon Lee* Division of Food Bioscience and Technology, College of Life Sciences and Biotechnology, Korea University 1 Uiseong Black Garlic Farming Association Abstract We investigated the antioxidative effect of black garlic extract (BGE) on paraquat (PQ)- induced oxidative stress in mice. A DPPH radical scavenging activity assay showed that BGE had potent free radical scavenging activity, comparable to that exhibited by vitamin C. Mice were administered with either vitamin C or two levels of BGE by oral gavage for 10 days, with PQ being injected intraperitoneally on day five. It was found that BGE reduced the liver enzyme levels induced by PQ injection in mice. The concentrations of plasma and hepatic malonedialdehyde were both significantly reduced in the BGE groups compared with the levels in the PQ group, whereas the hepatic superoxide dismutase and catalase activities were significantly increased in the BGE groups compared with the PQ group. These findings suggest that BGE has potent antioxidative activities in vivo and thus could prevent the oxidative stress induced by PQ injection in mice by two mechanisms: the induction of antixoxidative enzyme systems and direct scavenging of reactive oxygen species. Keywords: black garlic, antioxidant, oxidative stress, paraquat 서 생체내에너지대사과정은산소분자를이용한산화작용이끊임없이일어나며이과정중상당량의활성산소종이생성된다. 과량의활성산소는세포내에여러가지유해한결과를초래하는데, 예를들면, 지질과산화반응을촉진하여반응성이매우높은과산화물이생성되고, 이러한물질들은다양한 2 차산물로전환되어면역계손상등각종세포독성을나타낸다. 특히, 생체막의구성성분인인지질의불포화지방산을공격하여세포및조직의기능의저하를통해노화및성인병을유발하는것으로도알려져있다 (1). 이와같은활성산소의생성은다양한환경요인, 식이요인, 유전적요인에의해촉진되는데, 즉헤모글로빈이나티올기같은비단백성대사물질의자가산화과정, 흡연및오존의영향, paraquat, 사염화탄소등과같은독성화합물에의해서도많이생성되며이러한요인에노출이되면활성산소및산화적스트레스증가에따라노화및성인병발병위험이증가되는것으로보고되고있다 (2). *Corresponding author: Sung-Joon Lee, Division of Food Bioscience and Technology, College of Life Sciences and Biotechnology, Korea University, Seoul , Korea Tel: Fax: junelee@korea.ac.kr Received October 29, 2011; revised November 22, 2011; accepted November 22, 2011 론 건강한세포는대사과정중에생성된활성산소를중화하여자기스스로를보호하는방어기전이존재하는데, 이러한방어시스템은크게효소계와비효소계로구성되어있다. 즉, 효소계로서 superoxide dismutase(sod), catalase, glutathione peroxidase(gsh- Px) 등의항산화효소 (3,4) 들이활성산소를제가하는방법이있으며, 비효소계로서항산화활성이강한유기화합물인비타민 C, 토코페롤등에의해활성산소를제거하는메카니즘을가지고있어세포내활성산소생성을효율적으로제어하고있으나, 여러가지요인에의해세포내활성산소종이과다하게생성되거나생체내항산화시스템의활성이저해되어균형이깨어지면, 조직내산화적스트레스가증가하여노화및각종성인병발병의위험이증가하게된다. 따라서노화및성인병예방을위해서는생체및조직내산화적스트레스를효율적으로조절하는것이매우중요하므로다양한방법이연구및개발되고있는데, 항산화물질이포함되어있거나항산화효소계를활성화하는식품을이용한방법도주목받고있으며, 생체내에서이러한활성산소의생성을억제하거나, 생성된활성산소라디컬물질을소거하는식물유래항산화식품에대한연구가전세계적으로많이진행되고있다. Paraquat(PQ, N,N-dimethyl-4,4-bipyridinium dichloride: methyl viologen) 은 1958년영국에서개발하여 Gramoxone이라는상품명으로시판되어 1950년대광범위하게사용된제초제의일종으로미생물이나식물, 동물, 인간등에치명적인독성을나타내는것으로알려져있다. 이러한독성은 PQ가세포내의효소작용에의해 PQ 라디컬로변화되며그후의자동산화반응에서슈퍼옥사이 760

2 흑마늘추출물의항산화효과 761 드 (O 2 ) 의생성에기인하는것으로알려져있으며, 이러한자유라디컬은세포막파괴등여러생화학적경로를통해세포손상을유도하므로인체노출시에강력한독성을지니게되며 (5), 항산화실험에서산화적스트레스를유발하는물질로도널리사용되고있다. 백합과파속에속하는마늘 (Allium sativum L.) 은다년생채소로서강장, 강정식품으로널리이용되어왔으며, 특유의풍미가있고생리활성이우수하여조미료, 라면스프, 소스류등의가공식품의향신료및건강기능성식품으로광범위하게이용되어왔으며그수요가지속적으로증가하고있다. 마늘은주요성분인유기유황화합물의 -SH 기의강력한환원력으로인하여항산화활성이강하며, 이로인해세포의활성을촉진하고항암작용, 동맥경화예방, 항균작용, 항산화작용및중성지질및콜레스테롤수치를낮추는효능등여러가지건강기능성이보고되고있다 (6,7). 최근에는마늘을일정한온도와습도에서장기간숙성시켜만든흑마늘이제품화되고있는데흑마늘은일정한온도와습도를유지하는가공과정을통하여포도당, 과당, 자당및맥아당등유리당의함량이증가되고산도가낮아져서독특한맛을갖게되며, 마늘자체에함유된당과아미노산에의한갈변반응및중합반응으로인해흑색으로변화된다 (8). 이러한흑마늘은생마늘에비해아존성분이많이함유되어있어항혈액응고및뇌혈관, 콜레스테롤저하에도움이되며 (9), 셀레늄은항산화및항암효과, 심혈관계질환예방효과가있다고보고되고있다 (10). 이러한건강기능성에대한기대감으로인해흑마늘환, 흑마늘액기스, 흑마늘파우더등의여러가지가공제품들이유통되고있으며건강기능에대한연구도시작되어왔다. 항산화능과관련된연구로서제 2 형당뇨쥐를대상으로동결건조한마늘과흑마늘을섭취시킨결과항산화효소농도가증가하였고 (9), 흰쥐를대상으로 5 주간마늘분을섭취시킨결과지질과산화가억제되고항산화효소활성이증가하였으며 (11), 흑마늘분말첨가급여가고콜레스테롤흰쥐의지질대사에미치는영향등에대한연구가보고된바있으나흑마늘의생리활성규명및관련물질에대한연구를위해항산화작용에대한다양한연구가보다분명히규명되어야할필요성이있다. 따라서본연구는생쥐를이용한동물실험을통하여 PQ 에의한산화적스트레스에대해서, 흑마늘추출물의섭취가항산화효소계에어떠한영향을미치는지를관찰하고흑마늘추출물의항산화능력을측정함으로써흑마늘추출물의건강기능식품소재로써의가능성을검토하고자하였다. 재료및방법 실험동물및시약실험동물은샘타코 (BioKorea, Osan, Korea) 에서 6 주령의수컷 ICR 생쥐를실험동물용사료로 7 일간사육시킨후, 실온 o C, 습도 50-60%, 12 시간명암주기의사육조건에서사료와물을자유급여하였으며, 체중이 g 인생쥐를사용하였다. 동물실험프로토콜은고려대학교실험동물윤리위원회의승인을받은방법에따라수행하였다 ( 동물실험프로토콜번호 : KUIACUC ). 실험에사용한 vitamin C 와 PQ 는 Sigma Aldrich (St. Louis, MO, USA) 에서구입하였으며흑마늘추출물은의성흑마늘영농조합에서제공받았다. 실험동물처리실험동물군은각군당 ICR 계생쥐 8 마리씩무작위로배정하였으며, 무처리대조군, PQ 투여군, PQ + 흑마늘저농도 (BGL), PQ + 흑마늘고농도처리군 (BGH), 양성대조군인 PQ+vitamin C 처리군등 5 군으로나누어실험을수행하였다. 무처리대조군에는생리식염수를경구투여하였으며나머지실험군에는 PQ(25 mg/ kg, single dose) 를복강주사를통해투여하였는데, 흑마늘저농도군에는 90 ml/kg BW 의흑마늘추출물, 고농도에서는 180 ml/ kg BW 의흑마늘추출물, vitamin C 투여군은 400 mg/kg BW 의비타민 C 를 PQ 투여전 5 일, 투여후 5 일동안경구투여하여실험하였다. 흑마늘투여양은 65 kg 성인에게흑마늘추출물각각 90 ml(2 포 ) 및 180 ml(4 포 ) 에해당하는양이었다. 분석시료처리흑마늘추출물과 vitamin C 를 5 일간경구투여한후 24 시간절식시킨실험동물은심장에서혈액을채취한후혈중간기능지표효소의활성측정과혈액내지질과산화측정을위해사용하였고, 간조직은적출후간조직의일부분을취하여세번생리적식염수로세척하여혈액을제거한다음 80 o C 에보관하여간조직내지질과산화측정과 SOD, catalase, 글루타치온활성도의측정에사용하였으며조직의단백질함량은 bovine serum albumin 을표준물질로사용하여 Bradford(12) 방법에따라측정하였다. 혈청중간기능지표효소의활성도측정혈청 alanine aminotransferase(alt) 및 aspartate aminotransferase(ast) 활성도는쥐의혈액을채취하여 13,000 g 에서 20 분간원심분리하여혈청을분리한다음 Cobas C111(Roche, Basel, Switzerland) 를사용하여흡광도를이용하여측정하는방법을통하여측정하였다. 자유라디컬소거활성측정실험에사용된 DPPH(2,2-diphenyl-1-picrylhydrazyl) 는 Sigma Aldrich 제품을사용하였다. DPPH 라디컬소거능은 Blois(13) 의방법을응용하여 DPPH 라디컬에전자를공여함으로써라디컬을소거하는효과를 517 nm에서흡광도를측정하여계산한후나타내었다. 즉 0.15 mm DPPH solution(99.8% methanol) 을제조한후, 흑마늘추출물을 0.125, 0.25, 0.5, 1%(v/v) 의농도로증류수에희석하여만든시료 0.1 ml을 DPPH solution 2.9 ml에가하여교반한다음실온에서 30분간반응시킨후 517 nm 에서흡광도를측정하였다. 양성대조군으로항산화제인 vitamin C(100 µg/ml) 를, 증류수를대조군으로사용하여동일한방법으로측정하였다. 측정값은대조군의흡광도에대한흑마늘처리군의상대적흡광도 (%) 값을이용하여다음의공식에대입하여시료의전자공여능으로나타내었다. Antioxidant efficiency (%)= Ab Samples Ab Revision % Ab Standard 혈액과간조직중의과산화지질함량측정혈액과간조직내의지질과산화물의함량은 Ohkawa 등 (14) 의방법에의한 TBA 법을이용하여 sample 의 532 nm 에서의흡광도를이용하여생성된 malonedialdehyde(mda) 양을측정하였다. 혈액과간조직 1g 당생성된 MDA 를 µm 단위로나타내었다. 항산화효소계활성측정 SOD 활성은 SOD assay kit(dojindo Molecular Technologies, Tokyo, Japan) 를이용하고 catalase 활성도는 Catalase assay kit

3 762 한국식품과학회지제 43 권제 6 호 (2011) Table. 1 Experimental groups Group Treatment C No paraquat+h 2 O PQ Paraquat+H 2 O PQ+Vit C Paraquat+Vitamin C (400 mg/kg BW) PQ+BGL Paraquat+Black garlic extract (90 ml/kg BW) PQ+BGH Paraquat+Black garlic extract (180 ml/kg BW) PQ was injected in a single dose (25 mg/kg BW). Vitamin C and black garlic were administered orally before 5 days and after 5 days of PQ injection. Vitamin C was used as a positive control. Group C and PQ was received H 2 O instead of BGE (Abcam, Cambridge, UK) 을이용하여제조사의지시에따라각그룹당 3 번반복하여측정하였다. 글루타치온함량측정글루타치온 (glutathione, GSH) 함량은 GOH assay kit(biovision, San Francisco, CA, USA) 를이용하여제조사의지시에따라서각그룹당 3 번반복하여측정하였으며 1g 간조직당 µmol 의함량으로나타내었다. 통계처리모든실험은 3 반복으로측정하여측정치를 mean±sem 으로나타내었고실험결과의통계적유의성은 Student s t-test 를이용하여 p 값이 0.05 미만일때통계적으로유의성이있다고판단하였다. 결과및고찰 DPPH 라디컬소거능세포내자유라디컬은약 90% 가미토콘드리아에서일어나는산화적인산화과정에서일어나며그외에도식세포작용, 세포질중 xanthine oxidase 나 glutathione reductase 등의 flavoenzyme 에의한대사과정등여러가지생물학적반응에의해형성되며전자공여작용은이런산화성생물활성자유라디컬에전자를공여하여산화를억제하는항산화작용의주요한척도중하나이다 (15). DPPH 는짙은자색을띄는비교적안정한자유라디컬로항산화물질에의한전자공여에의해지질과산화연쇄반응에관여하는산화성자유라디컬의억제정도를측정할수있다 (16). 본실험에서사용한흑마늘추출물을증류수에희석하여농도별전자공여능을조사한결과는 Fig. 1 과같다. 흑마늘추출물은 0.125%(v/ v) 에서 1%(v/v) 까지의농도에서대조군인증류수와비교하여유의적인차이를나타내었으며 0.125% 의농도에서는, 흑마늘추출물의전자공여능은대조군에비해 0.125% 에서 10%, 1%(v/v) 의농도에서 50% 높게농도의존적으로증가하는경향을나타내었다. 또한 1% 의농도에서는양성대조군으로사용된 vitamin C(100 µg/ml) 보다도약 2 배정도의더높은전자공여효과를나타내었다. 마우스체중의변화 PQ 와흑마늘추출물의투여가마우스의체중에미치는영향을측정하기위하여 PQ 투여 5 일전과투여후 5 일동안 vitamin C, 흑마늘저농도와고농도를경구투여한후마우스체중의변화를측정한결과, PQ 투여군은투여전과비교하여평균체중이약 10% 정도감소하였으나, PQ 를투여하지않은대조군은유의적으로체중이증가하였으며, 흑마늘투여군은유의성을나타내지는않았지만평균체중이무처리대조군과유사하게증가 Fig. 1. DPPH radical scavenging activity of BGE. Radical scavenging activity was assessed as described in Materials and Methods using DPPH. H 2 O was used as a control for group C. The data are means±sem (n=3 per group). *p<0.05; **p<0.01; ***p<0.001 compared with PQ group. 하는경향을나타내었다 (Fig. 2A). 이실험결과를통해, PQ 에의한급격한산화적스트레스가마우스의체중감소를유발하였으나, 흑마늘추출물을투여하면 PQ 의산화적스트레스를감소시켜, 보호효과를지니면서체중의감소를억제한것으로보인다. 혈액내간기능지표측정흑마늘추출물의섭취가 PQ 의산화적스트레스에의해서유발된간손상에대한영향을검토하고자 PQ 투여전 5 일, 투여후 5 일동안흑마늘추출물을투여후, 혈청 AST, ALT 의활성을측정하였다 (Fig. 2B). PQ 투여군은무처리대조군에비해 AST 농도는약 2 배, ALT 농도는약 1.5 배의유의적인증가를보였으며, 흑마늘저농도와저농도처리군은대조군과비슷한수준의 AST 와 ALT 의값을나타내었다. 이결과는 PQ 의투여는산화적스트레스를증가시켜간조직손상을일으켜 AST 와 ALT 를증가시키지만, 흑마늘을투여하면 PQ 에의한산화적스트레스를감소시켜간조직손상을억제하고따라서혈중간조직손상지표인 AST 와 ALT 농도를감소시킨다는것을의미한다. 즉, 흑마늘투여는산화적스트레스에의한간조직손상을억제할수있음을보여주며, 저농도투여로유의적인효과를볼수있음을보여준다. 혈액과간에서의지질과산화물함량변화지질과산화반응은여러가지독성화합물이나약물에의한간손상발생의가장중요한기전으로써인정되어지고있고 (16), 이러한기전은세포내산화적스트레스의증가, 즉자유라디컬생성의증가및항산화적방어력의감소에의해야기된다고볼수있다 (17). 마우스에 PQ 를투여한후, 흑마늘추출물을경구투여하여산화적스트레스에대한회복효과를연구하기위해마우스의혈액과간조직에서지질과산화반응의부산물인 MDA 함량을 Thiobarbituric acid reactive substances(tbars) 정량법을이용하여측정하였다 (Fig. 3). 혈액에서흑마늘추출물저농도와고농도처리군은 PQ 처리군에비해 MDA 함량이유의적으로낮게나타났으며, 조직에서는흑마늘추출물고농도처리군에서 PQ 처리군에비해서 MDA 함량이 60% 정도낮게나타났다. 지질과산화는활성산소매개체가세포자체의국소적인방어작용을초

흑마늘추출물의항산화효과 763 Fig. 2 The effect of BGE on paraquat-induced toxicity in mice. (A) Mouse body weight, BGE and vitamin C were orally administrated for 10 days and PQ was injected on day 5.

days and injected with PQ on day 5. (B) Plasma AST and ALT levels. The data are means±sem (n=7 per group). *p<0.05; **p<0.01; ***p<0.001 compared with PQ group. Fig.

4 흑마늘추출물의항산화효과 763 Fig. 2 The effect of BGE on paraquat-induced toxicity in mice. (A) Mouse body weight, BGE and vitamin C were orally administrated for 10 days and PQ was injected on day 5. C, control; PQ, mice injected with PQ alone; PQ+VitC, mice administered vitamin C for 10 days and injected with PQ on day 5; PQ+BGL and PQ+BGH, mice administered low and high levels of BGE for 10 days and injected with PQ on day 5. (B) Plasma AST and ALT levels. The data are means±sem (n=7 per group). *p<0.05; **p<0.01; ***p<0.001 compared with PQ group. Fig. 3 The effect of BGE on MDA concentrations in the plasma and the livers in mice injected with PQ. MDA concentrations were quantified with TBARS assay. Group designation is the same as shown in Fig. 2. *p<0.05; **p<0.01; ***p<0.001 compared with PQ group. 과함으로써발생되는세포손상의주된형태이다. 이결과를통하여 PQ 의투여가혈액에서의지질과산화를유발하며흑마늘액기스의섭취가이러한지질과산화의생성에의한세포나조직의손상을억제함으로써산화적스트레스에대한방어작용을일으킨다고결론내릴수있다. 간조직내항산화효소활성변화급성간손상과정에서는글루타치온함량감소와대사활성체에의한지질과산화반응이일어난다. 지질과산화반응은간기능의부전을야기하는기본적인기전중의하나로중요시되어왔으며 (18), 이러한지질과산화반응에의한간손상예방과관련된기전은글루타치온대사와밀접하게관련되어있을뿐아니라 (19), 간조직내에서 superoxide 를제거하는 SOD, 그리고과산화수소를제거하는 catalase 와같이산소라디컬을제거하여최 종적으로지질과산화를감소시키는항산화효소들도산화적스트레스에의한세포나조직의손상의방어작용에중요한역할을담당한다 (20). 마우스의간조직에서 PQ 대사과정에서생성된 superoxide 라디컬의소거기능을가진 SOD 의활성도를측정하여단위 mg 단백질당 SOD 의활성도 (U) 로나타내었다 (Fig. 4A). 대조군과 PQ 군간에는유의적인차이가나타나지않았으며, 흑마늘추출물투여군은 PQ 를단독투여한군과비교하여저농도와고농도투여군모두 SOD 활성도가유의적으로증가하였다. Catalase 의활성은단위 g 단백질당 catalase 의활성도 (mu) 로나타내었으며 SOD 측정값과유사한경향을나타내었다 (Fig. 4B). 흑마늘투여군들은 PQ 투여군에비하여유의적으로 catalase 활성이증가하는경향을보였으며흑마늘고농도투여군은대조군과비교하여약 20% 정도의높은 catalase 활성을보였다. 위

764 한국식품과학회지제 43 권제 6 호 (2011) Fig. 4 The effect of BGE on the antioxidant enzyme activities and the plasma GSH levels in the livers of PQ-injected mice.

5 764 한국식품과학회지제 43 권제 6 호 (2011) Fig. 4 The effect of BGE on the antioxidant enzyme activities and the plasma GSH levels in the livers of PQ-injected mice. SOD (A) and catalase (B) activities were normalized with cellular protein concentrations. (C) The reduced form of GSH concentration. Group designation is the same as shown in Fig. 2. *p<0.05; **p<0.01; ***p<0.001 compared with PQ group. 의결과를통해서, 흑마늘추출물이간조직에서 SOD 의활성을증가시켜 PQ 의투여로인해서생성된 superoxide 라디컬을소거하고, SOD 의활성결과생성된과산화수소를 catalase 의활성유도를통해서제거함으로써 PQ 에의해유발된산화적스트레스로부터간을보호한것으로사료된다. 간조직내글루타치온함량측정간조직내글루타치온함량을측정하여 1g 간조직당 µmol 농도로나타내었다 (Fig. 4C). 글루타치온함량은대조군에서가장높게나타났으며, 흑마늘추출물투여군이 PQ 군보다농도의존적인경향으로높은글루타치온함량을나타내었지만 p<0.05 수준에서의유의성은나타나지않았다. 이실험결과, 흑마늘추출물의투여가 PQ 의체내유입에따른해독작용에사용된 GSH 의원활한환원을촉진한것으로추측되며따라서흑마늘추출물 GSH 의함량을증가시켜산화적스트레스에대해서간을보호하는기능을지닌다고추측할수있다. 직에서항산화효소인 SOD, catalase 활성을유의적으로증가시켰다. 또한, 흑마늘추출물은자유라디컬에의해생성된지질과산화의최종산물인 MDA 의함량을혈액과간조직에서유의적으로감소시키는효과를보였으며, 산화적스트레스에의한방어작용을하는 GSH 농도를유의적으로증가시켰다. 이와같이흑마늘추출물은항산화관련효소의활성도를증가시키고생체내에서내인성항산화물질의합성능력을강화시킴으로써산화적스트레스로인한생체손상에대한회복작용을향상시킨다고결론내릴수있으며, 항산화건강기능식품으로써의기능성이기대된다. 감사의글 본연구는보건산업진흥원이지원하는건강기능식품센터사업 (A ) 과산림청산림과학특정연구사업 (S120911L130110) 의지원을통해수행되었습니다. 요 약 문 헌 본연구에서는 PQ 의독성으로유발된산화적스트레스에대한흑마늘추출물의항산화활성효과를검정하여건강기능식품으로써의흑마늘추출물기능성을알아보기위해흑마늘추출물의전자공여능을측정하였으며마우스를이용하여 PQ 투여전 5 일, 투여후 5 일간흑마늘추출물을경구투여한후, PQ 독성에의한체중변화관찰과혈중간기능지표인 AST 와 ALT 값을측정하였다. 또한간조직에서 SOD, catalase 의활성과 GSH, MDA 함량등을측정하여비교함으로써흑마늘추출물의황산화활성성분이 PQ 의독성에대한회복작용그리고지질과산화와의상호관련성을검토하였다. 흑마늘추출물의투여는간조 1. Rubin H. Cell aging in vivo and in vitro. Mech. Aging Dev. 98: 1-35 (1997) 2. Kelner MJ, Alexander NM. Inhibition of erythrocyte superoxidedismutase by diethyldithiocarbamate also results in oxyhemoglobin-catalyzed glutathione depletion and methemoglobin production. J. Biol. Chem. 261: (1986) 3. Scandalios JG. Oxygen stress and superoxide dismutases. Plant Physiol. 101: 7-12 (1993) 4. Harris ED. Regulation of antioxidant enzymes. FASEB J. 6: (1992) 5. Suntres ZE. Role of antioxidants in paraquat toxicity. Toxicology 180: (2002) 6. Lee SJ, Kim JH, Kim MJ, Yoon SM, Jeong JC, Sung NJ. Effect

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Journal of Life Science 2011, Vol. 21. No μ μ

Journal of Life Science 2011 Vol. 21. No. 8. 1120~1126 ISSN : 1225-9918 DOI : http://dx.doi.org/10.5352/jls.2011.21.8.1120 μ μ μ α β Journal of Life Science 2011, Vol. 21. No. 8 1121 μ μ 1122 생명과학회지 2011,