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1 Korean J Food Cook Sci Vol. 32, No. 6, pp. 818~827 (2016) pissn eissn 천마추출물이알코올을투여한흰쥐의간기능및알코올대사에미치는영향 박순자 강주현 공현주 1 황수정 1 장정현 2 양경미 1 대구한의대학교대학원한방식품학과, 1 대구한의대학교한방식품조리영양학부, 2 부산동의과학대학교피부미용계열 Effects of Gastrodiae rhizoma on the Liver Function and Alcohol Metabolism in Alcohol Treated Rats Soon-Ja Park Ju-Hyeon Kang Hyun-Joo, Kong 1 Su-Jung Hwang 1 Jung-Hyeon Jang 2 Kyung-Mi Yang 1 Department of Herbal Food Science, Graduate School of Daegu Haany University, Gyeongsan 38610, Korea 1 Faculty of Herbal Food Cuisine and Nutrition, Daegu Haany University, Gyeongsan 38610, Korea 2 Division of Cosmetology, Dong-Eui Institute of Technology, Busan 47230, Korea Abstract Purpose: This study was conducted to investigate the effect of freeze-dried Gastrodiae rhizoma powder (GR) on the liver function and alcohol metabolism in alcohol treated rats. Methods: The rats were administered various concentrations of GR (100 mg, 200 mg, 300 mg/kg B.W.) for 3 days 1 hour before 50% (v/v) ethanol (3 g/kg B.W.) administration. Two tests focusing on liver function and alcohol metabolism in acute alcohol treatment were carried out. Results: Glutamic oxaloacetic transaminase activity was significantly increased by alcohol treatment, and was decreased by 100 mg GR administration. Acute ethanol treatment led to significant increase in alcohol and acetaldehyde levels of serum and liver. However, 100 mg GR administration led to a significant reduction in increased alcohol level in the serum with decreased alcohol dehydrogenase (ADH) activity and increased acetaldehyde level in liver was significantly reduced by three levels. Conclusion: These results suggest that GR can be effective in enhancing liver function and alcohol metabolism in the alcohol-treated rats. Studies on the appropriate dosage of GR should further be developed to treat alcohol detoxification and stimulate liver function. Key words: Gastrodiae rhizoma, ethanol, liver function, acetaldehyde, ADH Ⅰ. 서론 현대사회와산업의급격한환경변화로인한스트레스로음주량이증가되면서음주로인한경제적손실은심각한것으로나타났다. 급만성의알코올섭취는숙취와두통으로인한사회활동장애를일으킬뿐만아니라신체적으로간, 위, 식도, 심장이외에사고력이상, 기억장애및말초신경변증과같은신경계손상을유발한다. 특히, 섭취된알코올이주로해독되는간조직에서심각한손상을일으키므로급만성알코올섭취로인한질환중에서알코올성간질환 (alcoholic liver disease) 의발생빈도가가장높은것으로보고되고있다 (Lieber CS 1993). 섭취된알코올의 20% 는위장관에서흡수된후 alcohol dehydrogenase(adh) 에의해대사되며, 나머지약 80% 는소장상부에서확산작용으로흡수된후간조직으로이동되어대사된다. 그리고일부의대사되지않은알코올은폐나소변및땀으로배설된다 (Morris R 1984, Lieber CS & Decarli LM 1991). 알코올이 acetaldehyde 로산화되는대사체계는 ADH, microsomal ethanol oxidizing system (MEOS) 의 cytochrome P450 2E1(CYP2E1) 및 catalase 효소에의해주로간조직에서이루어지므로간은알코올대사에중요한작용기관으로작용한다 (Lands WE 1998, Das SK & Vasudevan DM 2007). ADH 는알코올에대한 Km 의농도가 mm 으로체내알코올의농도가낮을때작용하며, MEOS 는 ethanol 에대한 Km 의농도가약 10.0 mm 으로알코올의농도가높을때작용하는것으로 Corresponding author: Kyung-Mi Yang, Department of Herbal Food Science, Graduate School of Daegu Haany University, 1, Hanuidae-ro, Gyeongsan-si, Gyeongbuk 38610, Korea ORICID: Tel: , Fax: , jiboosin@dhu.ac.kr Korean Society of Food and Cookery Science This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License ( nses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

2 Korean J Food Cook Sci 천마추출물이알코올을투여한흰쥐의간기능및알코올대사에미치는영향 819 알려져있다 (Jung BS 1991). 간조직에서알코올대사에중요한경로는 ADH 와 MEOS 체계이며, catalase 의역할에대해서는아직까지명확하게밝혀지지않고있다 (Gill K 등 1996). 알코올의산화로생성된 acetaldehyde 는 acetaldehyde dehydrogenase(aldh) 에의해산화되며, 산화물의일부는혈액으로이동된다 (Jung BS 1991). 급만성적으로과잉의알코올을섭취할경우간조직에서알코올대사효소체계의활성변화로알코올대사가저해되어알코올, acetaldehyde, 대사과정중생성되는유해대사산물, 활성산소 (reactive oxygen radicals, ROS) 및 nicotinamide adenine dinucleotide(nadh) 의과잉생성등으로부터야기되는다양하고복잡한기전을통하여알코올성지방간, 간염, 간경화증및간암을유발해사망을초래하게된다 (Tsukamoto H 등 1986, Lieber CS & Decarli LM 1991, Keshavarzian A 등 1994, Kim IH 2008). 따라서알코올성간손상정도는알코올및 acetaldehyde 의대사효소의활성조절을통한이들의분해정도에따라영향을받는다. 현재까지체내에서알코올과 acetaldehyde 농도를빠르고안전하게낮추어숙취해소와간기능회복에도움을준다고밝혀진천연물로는헛개나무와천마혼합추출물 (Jeon TW 등 2002), 오미자농축액 (Han CK 등 2014), 민들레즙 (Noh KH 등 2009), 보이차 (Park SH 등 2004) 등이보고되었다. 난초과 (Orchidaceae, 蘭草科 ) 에속하는다년생초본 ( 多年生草本 ) 인천마 ( 天麻, Gastrodiae rhizoma) 는약성이평무독 ( 平無毒 ) 하며미 ( 味 ) 는감 ( 甘 ) 한것으로알려져있다 (Kwon HJ 등 2014). 천마의효능에관한국내연구로는뇌혈류순환개선과항고혈압 (Park SH 등 2005, Hong HD 등 2007), 혈당과혈청지질성분조절 (Kim KI 등 2003, Yang KM 2008), 항산화와항염증반응을통한간보호 (Kim HJ 등 2016), 간과뇌조직의불포화지방산농도조절 (Park PS & Park MY 2001) 및알코올대사활성 (Shim EJ 2006) 등이활발히진행되고있다. 위에서살펴본바와같이천마는항산화력을기반으로혈류량과혈압을조절하고지질대사를개선하여간조직을보호하는것으로보고되고있지만알코올성간질환에대한천마를연구한보고는거의없는실정이다. 이에본연구에서는동결건조시킨천마분말이흰쥐의간기능과알코올대사에미치는영향을구명하여알코올성간손상에대한건강기능성식품의소재로서천마의활용방안을탐색하고자한다. Ⅱ. 재료및방법 1. 실험재료및시약천마는전라북도무주에서무주생천마를구입하여이용하였다. 실험동물의사료는일반배합고형사료 (Jeil Feed Co., Daejeon Korea) 을사용하였다. 간기능은 glutamic oxaloacetate transaminase(got) 와 glutamic pyruvate transaminase(gpt) 활성으로측정하였으며, 측정용 kit 는 Asan Pharma Co.(AM101, Seoul, Korea) 제품으로구입하였다. 알코올대사를측정하기위한알코올 (N7160, Sigma-Aldrich Co., St. Louis, MO, USA) 과 acetaldehyde(ab112113, Abcam, Cambridge, UK) 농도및 ADH(K , Biovision, Milpitas, CA, USA) 와 ALDH(K , Biovision) 의효소활성은상업용으로제조된측정용 kit 를구입하여분석하였다. 2. 천마동결건조분말제조생천마는일반음용수로세척하여흙과먼지등의이물질을제거한다음수세미로껍질을벗겨내고깨끗이씻었다. 그런다음 5-10 mm 두께로절편한천마를 100 C 에서 40분간증자한것을 80 C에서 8시간건조한다음, 분쇄한시료의 10배량의증류수를넣고 3시간동안 85 C 에서추출하였다. 그런다음회전식증발농축기 (Rotavapor R-210, Buchi, Flawil, Switzerland) 를이용하여감압농축하였다. 그리고동결건조기 (FD 5510 SPT, Ilshin, Seoul, Korea) 에서동결건조시킨후 -70 C의초저온냉동고 (MDF- U52V, SANYO Electric Co., Ltd., Osaka, Japan) 에보관한다음시료로사용하였다. 만들어진천마추출물동결건조분말은생리적식염수를이용하여 1%, 2%, 3% 로만든다음실험동물에게경구투여하였다. 3. 실험동물및알코올투여실험동물은 g 정도의생후 10주령이된 Sprague Dawley(SD) 종수컷흰쥐로 ( 주 ) 효창사이언스 (Daegu, Korea) 에서분양받았다. 먼저 2주동안일반배합고형사료로환경에적응시킨다음체중에따라각실험군당 7 마리씩 5군으로각각의 cage에분리하고완전임의배치하여사육하였다. 사육환경조건으로온도는 22±1 C, 상대습도는 65±5%, 명암은 12시간주기 (09:00-21:00) 로조절하였다. 물은임의로섭취하도록하였다. 실험군은 Table 1과같이정상군 (NC), 알코올단독투여군 (EC) 및체중 kg 당각각 100 mg(eg100), 200 mg(eg200), 300 mg(eg300) 천마분말과알코올투여군으로나누었다. 천마분말과알코올은생리적식염수에희석시켜이용하였다. 천마분말시료와알코올투여는매일일정한시간에 3일동안위관경구투여 (oral administration) 로이루어졌으며, 천마분말 (100 mg/kg B.W., 200 mg/kg B.W., 300 mg/kg B.W.) 을투여한지 1시간이지난다음생리식염수로희석된에탄올 (50%, v/v) 을체중 kg당 3 g을투여하였다. 이때정상군은알코올대신에생리적식염수만을경구투여하였다. 마지막 3일째되는날은알코올을투여한지 1시 ; 32(6):

3 820 박순자등 Korean J Food Cook Sci Table 1. Classification of experimental groups Gruop 1) Ethanol (g/kg B.W.) Gastrodiae rhizoma (mg/kg B.W.) NC - - EC 3 - EG EG EG ) NC: 0.9% saline + 0.9% saline; EC: ethanol 3 g/kg B.W % saline; EG100: ethanol 3 g/kg B.W. + G. rhizoma 100 mg/kg B.W.; EG200: ethanol 3 g/kg B.W. + G. rhizoma 200 mg/kg B.W.; EG300: ethanol 3 g/kg B.W. + G. rhizoma 300 mg/kg B.W. 간이지난다음희생시켜시료를채취하였다. 4. 시료채취 천마분말과알코올을급성투여한흰쥐는 CO 2 (Shinsung, Gimhae, Korea) 로가볍게마취시켜개복한즉시복부대동맥에서혈액을채취하였다. 혈액은실온에서약 30 분간방치한후 3,000 rpm 에서 15 분간냉장상태에서원심분리 (VS-550, Vision Scientific Co., Daejeon, Korea) 하여얻은혈청을일정량씩나눈다음액체질소탱크 (XC Millennium 20, Chart/Mve, Buffalo, MN, USA) 에서급속동결한후분석전까지 -70 C 에서초저온냉동고 (SANYO Electric Co., Ltd.) 에서냉동보관하였다. 간조직의일부를취해서조직균질기 (HS-30E, Daihan Scientific, Wonju, Korea) 를사용하여냉장상태에서 1.15% KCl 완충용액을제조하여 10%(w/v) 마쇄균질액을만든다음실험에사용하였다. 5. 실험방법급성알코올투여에의한간손상지표로혈청중 GOT 와 GPT 활성은각기질과효소반응을이용한비색법에의해제조된 GOT와 GPT 활성측정용 kit를이용하여측정하였으며, 활성은 Karmen unit/ml로나타내었다. 혈청과원심분리로얻은간조직균질액의알코올농도는 96-well plate를이용한알코올측정용 kit로반응시킨다음 microplate reader(n10583, Thermo Scientic, Waltham, MA, USA) 에서측정하였다. Acetaldehyde 농도는 acetaldehyde 가 ALDH에의해산화되어 acetic acid가만들어지는과정에서생성된 NADH의농도를측정하는원리로제조된 acetaldehyde quantification assay kit를사용하여측정하였다. ADH와 ALDH 활성은 NAD와반응하여생성된 NADH의농도를측정하는원리로제조된분석용 kit를이용하여 450 nm에서측정하였다. ADH와 ALDH 활성은분당 ml에서생성된 NADH 양을 nmol로나타내었다. 6. 동물실험윤리승인본연구는동물보호법 ( 제정 1991년 5월 31일법률제 4397호, 일부개정 2008년 2월 29일법률제8852호 ) 을근거하여대구한의대학교동물실험윤리위원회에의해승인받아수행되었다 (Approval Number: DHU ). 7. 통계처리실험을통하여얻어진자료는 SPSS Statistics(ver. 15.0, SPSS Inc., Chicago, IL, USA) 를이용하여분석하였으며그결과는평균 ± 표준편차로표시하였다. 실험결과는일원배치분산분석 (one way analysis of variance) 을한후 Duncan's multiple range test에의해 p<0.05 수준에서각실험군의평균치의통계적유의성을검정하였다. 천마분말과알코올투여로인한간손상지표, 알코올과 acetaldehyde 농도및 ADH와 ALDH 활성과의상관성은 Pearson's correlation coefficient(r) 를구한뒤 p<0.05 수준에서유의성을검증하였다. Ⅲ. 결과및고찰 1. 혈청중의 GOT 와 GPT 활성천마분말과알코올을투여한흰쥐의간손상의지표인자로알려진혈청중의 GOT와 GPT 활성에미치는영향은 Fig. 1과같다. GOT 활성은 NC군의 251.2±31.3 unit 에비해 EC군은 313.8±13.0 unit로알코올급여로약 24.9% 의증가를보였다. 천마분말을급여한 EG100군은 288.0±36.6 unit, EG200군은 311.3±40.8 unit, EG300군은 309.1±34.3 unit의 GOT 활성을보여 EC군에비해각각 8.2%, 0.80%, 1.50% 의감소를보였으나유의적인차이는없었다. 또한 GPT 활성은 NC군은 186.9±28.7 unit, EC군은 187.3±30.4 unit, EG100군은 195.9±14.1 unit, EG200군은 207.9±29.7 unit, EG300군은 188.8±14.3 unit로모든군간에유의적인차이는없었다. 따라서알코올투여로증가된 GOT 활성으로볼때알코올성간손상을받은것으로판단되며, 이때유의적이지는않지만낮은활성을보인 EG100군에서알코올성간손상에대해보호받은것으로추측된다. 급만성의과다음주는간조직의섬유화나불포화지방산조성비의불균형에의한세포막의구조및기능의변화를유발하게된다. 그결과, 간손상이진행되면서혈중알코올, acetaldehyde, GOT 및 GPT의수치가상승되는것으로여러연구자들에의해보고되었다 (Kwon YO 2003, Lee SY 등 2008, Choi GH 등 2011, Kim HJ 등 2016). 이상태가지속이될경우에는광범위한간조직의괴사와염증으로유발되는알코올성간염상태에서재생반응인섬유화과정을거치면서알코올성간경변증 2016; 32(6):

4 Korean J Food Cook Sci 천마추출물이알코올을투여한흰쥐의간기능및알코올대사에미치는영향 821 Fig. 1. Effects of Gastrodiae rhizoma on GOT and GPT activities in serum of alcohol treated rats. NC: 0.9% saline + 0.9% saline; EC: ethanol 3 g/kg B.W % saline; EG100: ethanol 3 g/kg B.W. + G. rhizoma 100 mg/kg B.W.; EG200: ethanol 3 g/kg B.W. + G. rhizoma 200 mg/kg B.W.; EG300: ethanol 3 g/kg B.W. + G. rhizoma 300 mg/kg B.W.. The results indicate the mean±sd of 7 rats per each group. Means with different letters (a,b) among group are significantly different by Duncan's multiple range test (p<0.05). NS: not significant. 각약 26.0% 와 29.0% 의억제를보여알코올성간독성의예방에생약제가유용하다는결과를밝혔다. 또한 Kim HJ 등 (2016) 은 lipopolysaccharide 의복강투여로유발된생쥐의간손상에대해천마및 6 번증숙발효천마를체중 kg 당 200 mg 투여하였을때 GOT 와 GPT 의활성이유의적으로 (p<0.001) 감소되었고, 그결과천마및증숙발효천마분말의간보호효과를밝혔다. 그러나본연구에서는알코올의투여로증가된 GOT 의활성에대해유의적이지는않지만 100 mg 의천마분말의투여로경미한 GOT 활성억제효과가있다는결론을얻을수있었다. Fig. 2. Correlation between serum alcohol level and GOT activity. 으로발전하게된다 (Kim IH 2008). 본연구에서도 Fig. 2 와같이혈청중의알코올농도가높을수록 GOT 활성이높은양의상관관계 (r=0.531, p<0.01) 를보임에따라서체내알코올농도가높을수록간조직의손상이심한것으로보여진다. Choi GH 등 (2011) 은체중 kg 당 3 g 의알코올 (50%, v/v) 을투여한지 8 시간이지난후에 GOT 와 GPT 활성을측정한결과, 정상군에비해각각약 81% 와 69% 의증가를보여알코올투여로간손상이일어났다고보고하였다. 그러나헛개나무열매추출물이나헛개나무를포함한 7 개의식품혼합물을투여했을때 GOT 와 GPT 활성은각 2. 혈청중의알코올수준천마분말과알코올을투여한흰쥐의혈청중의알코올농도는 Fig. 3과같다. 혈청 dl 당알코올농도는 NC 군의 34.05±7.30 mg에비해 EC군은 66.16±7.54 mg으로알코올의투여로 94.1% 증가된농도를보였다 (p<0.05). 그러나천마분말을급여한 EG100군은 53.18±4.86 mg, EG200군은 65.88±5.52 mg, EG300군은 86.90±10.08 mg으로천마분말의급여량과비례해서혈중알코올농도가증가되었다 (p<0.05). 또한 EC군에비해 EG100군에서는유의적으로 19.6% 의낮은알코올농도를보였으나, EG300 군에서는오히려 31.4% 의증가를보였다 (p<0.05). 알코올대사는 70 kg인성인남성의간조직에서보통한시간에체중 kg 당 100 mg 정도이루어지지만 (Choi GH 등 2011), 혈중알코올농도는알코올도수, 음주속 ; 32(6):

5 822 박순자등 Korean J Food Cook Sci 도및위내음식물의양과종류등에의해영향을받게된다. 그러나알코올의가수분해활성을가진천연물의공급으로혈중알코올농도를낮출수있다는사실이국내연구자들에의해밝혀졌다. Han CK 등 (2014) 과 Hwang Fig. 3. Effects of Gastrodiae rhizoma on alcohol level in serum of alcohol treated rats. NC: 0.9% saline + 0.9% saline; EC: ethanol 3 g/kg B.W % saline; EG100: ethanol 3 g/kg B.W. + G. rhizoma 100 mg/kg B.W.; EG200: ethanol 3 g/kg B.W. + G. rhizoma 200 mg/kg B.W.; EG300: ethanol 3 g/kg B.W. + G. rhizoma 300 mg/kg B.W.. The results indicate the mean±sd of 7 rats per each group. Means with different letters (a d) among group are significantly different by Duncan's multiple range test (p<0.05). SJ 등 (2010) 은알코올 (4 g/kg B.W.) 을투여한흰쥐에게오미자추출물및한약재추출물이함유된음료를급여한결과정상군에비해시간이경과할수록혈액에서의알코올농도감소속도가빠른것으로확인되었다고보고하였다. 또한본실험에서시료로이용한천마가함유된생약제혼합물을알코올 (3 g/kg B.W.) 을투여한흰쥐에게급여하였을때, 혈청중알코올농도가빠르게감소하고혈청중최고농도에도달하는시간및사다리꼴곡선하면적이 p<0.05 수준에서각각 33% 및 21% 의유의적인감소를보였다 (Jeon TW 등 2002). 그리고천마에서추출한저분자및다당체분획물도혈중알코올농도의수치를유의적으로낮추는것으로보고되었다 (Shim EJ 2006). 본연구에서도알코올을투여한흰쥐에게천마를급여하였을때, 혈청중의알코올농도는 100 mg 의천마분말투여군에서 19.6% 의감소효과가나타났다. 그러나 300 mg 투여군의혈중알코올농도는오히려증가현상을보였다. 그러나하루복용량은 20 g 을넘지않도록하고독성이나부작용과중독이있다는연구보고 (Kim YH & Seo BI 2012) 로볼때알코올과천마의상호작용에대한구명을통한음주시적절한천마복용량에대한연구가좀더필요할것으로보여진다. 3. 혈청과간조직중의 ADH 활성도천마분말과알코올을투여한흰쥐의혈청과간조직중의분당 ADH 활성은 Fig. 4와같다. 혈청중의 NC군과 EC군의 ADH 활성은각각 0.96±0.16 nmol/ml과 1.04 Fig. 4. Effects of Gastrodiae rhizoma on ADH activities in serum and liver of alcohol treated rats. NC: 0.9% saline + 0.9% saline; EC: ethanol 3 g/kg B.W % saline; EG100: ethanol 3 g/kg B.W. + G. rhizoma 100 mg/kg B.W.; EG200: ethanol 3 g/kg B.W. + G. rhizoma 200 mg/kg B.W.; EG300: ethanol 3 g/kg B.W. + G. rhizoma 300 mg/kg B.W.. The results indicate the mean±sd of 7 rats per each group. Means with different letters (a,b) among group are significantly different by Duncan's multiple range test (p<0.05). 2016; 32(6):

6 Korean J Food Cook Sci 천마추출물이알코올을투여한흰쥐의간기능및알코올대사에미치는영향 823 ±0.13 nmol/ml 로두군간에유의한차이는없었다. EC 군에비해 EG100 군은 1.26±0.104 nmol/ml, EG200 군은 0.91±0.21 nmol/ml, EG300 군은 1.00±0.12 nmol/ml 으로 EG100 군에서 21.2% 의 ADH 활성증가를보였다 (p<0.05). 간조직중의 ADH 활성은 NC 군의 50.84±3.15 nmol/ml 에비해알코올을투여한모든군에서 p<0.05 수준에서유의적으로낮은활성을보였으며, 알코올투여군들간에유의적인차이는없었다. 알코올투여군의활성치는 EC 군은 30.83±2.26 nmol/ml, EG100 군은 26.80±3.89 nmol/ml, EG200 군은 26.65±4.52 nmol/ml, EG300 군은 30.51±4.67 Fig. 5. Correlation between serum alcohol level and serum ADH activity. nmol/ml 이었다. 알코올을섭취한후에는 1 차적으로 ADH, microsome 의 CYP2E1 및 catalase 의활성이시간이경과하면서유도되어 acetaldehyde 로산화된다. 소화관에서는주로 ADH 에의해알코올분해가활발하게나타나고, 분해되지않은일부의알코올은호흡기를통해배설되면서혈중알코올농도가낮아지게된다 (Lieber CS 등 1994, Lieber CS 2004, Das SK & Vasudevan DM 2007, Chung YI 등 2009). Son JC 등 (2012) 과 Park SH 등 (2004) 은흰쥐 (5 g/kg B.W.) 와생쥐 (5.5 g/kg B.W.) 에게알코올을경구투여하였을때비투여군에비해혈중알코올농도는높았으나, ADH 활성은감소되었다고보고하였다. 본연구에서도 Fig. 5 에서처럼혈청중의알코올의농도가높았고, 간조직중의 ADH 활성은낮은음의상관관계 (r=-0.593, p<0.01) 로나타났다. 알코올대사의활성화로혈중알코올농도와알코올대사과정중생성되는유해대사산물및활성산소등의이차대사산물을감소시키는기전을통하여알코올성간손상을억제시키는활성물질에대한연구가활발히이루어지고있다. In vivo 실험에서는 Park SH 등 (2004) 은보이차추출물의급여로간조직에서 22.7% 의 ADH 활성이유도되어 87.3% 의혈중알코올농도감소효과를얻었다고하였다. 그리고 In vitro 실험에서는천마에서추출한다당체분획물과천마열수추출물이 ADH 활성을촉진하고, 천마저분자및다당체분획물이투여농도에비례해서 ADH 의활성을증가시키는경향을보였다 (Shim Fig. 6. Effects of Gastrodiae rhizoma on acetaldehyde levels in serum and liver of alcohol treated rats. NC: 0.9% saline + 0.9% saline; EC: ethanol 3 g/kg B.W % saline; EG100: ethanol 3 g/kg B.W. + G. rhizoma 100 mg/kg B.W.; EG200: ethanol 3 g/kg B.W. + G. rhizoma 200 mg/kg B.W.; EG300: ethanol 3 g/kg B.W. + G. rhizoma 300 mg/kg B.W.. The results indicate the mean±sd of 7 rats per each group. Means with different letters (a-c) among group are significantly different by Duncan's multiple range test (p<0.05) ; 32(6):

7 824 박순자등 Korean J Food Cook Sci EJ 2006). 따라서천마는알코올과 acetaldehyde 의대사에관여하는효소계를활성화시켜알코올로부터생체를보호할수있는기능성식품으로서가치가있을것으로판단된다. 본연구에서도알코올단독투여군에비해 100 mg 의천마분말을급여하였을때혈청중의 ADH 활성의유도로알코올농도를낮춘것으로보여진다. 4. 혈청과간조직중의 acetaldehyde 수준 천마분말과알코올을투여한흰쥐의혈청과간조직중의 acetaldehyde 농도는 Fig. 6 과같다. 혈청중의 acetaldehyde 농도는 NC 군의 4.61±0.60 µm 에비해 EC 군은 6.45±0.34 µm 로알코올투여로 39.9% 의유의미한증가를보였다 (p<0.05). 그리고 EG100 군, EG200 군, EG300 군은각각 6.25±1.32 µm, 6.61±1.13 µm, 7.01±0.93 µm 로천마분말의급여에의한혈청중의 acetaldehyde 농도의감소효과는없었다. 간조직중의 acetaldehyde 농도는 NC 군의 7.52± 0.59 µm 에비해 EC 군에서는 10.61±0.49 µm 로 p<0.05 수준에서알코올투여로 41.0% 유의미한증가를보였다 (p<0.05). 그러나 EG100 군, EG200 군, EG300 군의 acetaldehyde 농도는각각 8.80±1.03 µm, 8.40±1.09 µm, 8.37±0.67 µm 로 EC 군에비해각각 17.1%, 20.9%, 21.2% 가감소되었으며 (p<0.05), 천마분말의급여량에의존하여 acetaldehyde 농도의감소효과를보였다. 알코올성간손상은알코올자체, acetaldehyde, 알코올과 acetaldehyde 의대사과정중에생성된 ROS 및 NDAH /NAD 의비증가에따른대사장애에의해일어난다 (Lieber CS & Decarli LM 1991). 이중알코올보다독성이더강한물질로알려진 acetaldehyde 의농도가혈액에서높아지게되면두통과숙취를유발하고심한경우에는심혈관계및알코올성간손상질환으로사망하게된다. 본연구에서도 Fig. 7 과같이혈청 (r=0.629, p<0.01) 과간조직중 (r=0.364, p<0.05) 의 acetaldehyde 농도는 GOT 활성과양의상관관계를보이고있어 acetaldehyde 농도가가장높은 EC 군에서가장간손상이심할것으로판단 된다. 그러나인동초, 약쑥및들국화로만든하향주 (Jung HK 등 2008), 작약, 당귀및천궁혼합추출물 (Hwang SJ 등 2010), 민들레즙 (Noh KH 등 2009), 헛개열매추출 (Ko BS 등 2006), 눈꽃동충하초추출물 (Kim JM 등 2008), 천마분획물 (Shim EJ 2006) 등은 ALDH 효소의활성을유도하여 acetaldehyde 를분해시킴으로써알코올성간손상이나숙취에의한불쾌감을해소하고항산화력을높이는것으로보고되고있다. 특히, 천마추출물과분리한천마다당체는 in vitro 와 in vivo 실험에서 ALDH 의활성을증진시켜 acetaldehyde 의분해를촉진시키는효능이있는것으로나타났다 (Shim EJ 2006). 본연구에서도알코올투여로증가된 acetaldehyde 농도는혈청에서는천마의공급에따른유의적인차이는없었으나, 간조직에서는천마분말의급여량에의존적으로유의적인개선효과를보였다. 5. 혈청과간조직중의 ALDH 활성알코올과천마분말을투여한흰쥐의혈청과간조직에서의 1분당 ALDH 활성은 Fig. 8과같다. 혈청중의 ALDH 활성은 NC군의 13.58±3.37 nmol/ml에비해 EC군은 115.9% 증가된 29.33±2.39 nmol/ml의활성을보였다 (p<0.05). 그리고천마를급여한 EG100군은 22.86±1.42 nmol/ml, EG200군은 19.01±0.03 nmol/ml, EG300군은 24.53±1.87 nmol/ml의 ALDH 활성으로 NC군에비해높았다. 그러나 EC군에비해각각 22.1%, 35.2%, 16.4% 의낮은활성을보였다 (p<0.05). 간조직중의 ALDH 활성은 NC군은 12.06±0.30 nmol/ml에비해 EC군은 20.69± 0.29 nmol/ml로알코올투여로인해 71.5% 의높은활성을보였다 (p<0.05). 그러나 EG100군과 EG300군의 ALDH 활성은각각 16.56±1.40 nmol/ml와 13.81±1.21 nmol/ml로 EC군에비해 p<0.05 수준에서유의적으로낮은반면에, EG200군은 22.16±1.84 nmol/ml로유의적으로높은활성을보였다 (p<0.05). Fig. 7. Correlation between serum GOT activity and serum acetaldehyde (A) and hepatic (B) acetaldehyde level. 2016; 32(6):

8 Korean J Food Cook Sci 천마추출물이알코올을투여한흰쥐의간기능및알코올대사에미치는영향 825 Fig. 8. Effects of Gastrodiae rhizoma on ALDH activities in serum and liver of alcohol treated rats. NC: 0.9% saline + 0.9% saline; EC: ethanol 3 g/kg B.W % saline; EG100: ethanol 3 g/kg B.W. + G. rhizoma 100 mg/kg B.W.; EG200: ethanol 3 g/kg B.W. + G. rhizoma 200 mg/kg B.W.; EG300: ethanol 3 g/kg B.W. + G. rhizoma 300 mg/kg B.W.. The results indicate the mean±sd of 7 rats per each group. Means with different letters (a-e) among group are significantly different by Duncan's multiple range test (p<0.05). Fig. 9. Correlation between serum ALDH activity and serum alcohol (A) and acetaldehyde (B) level, and between hepatic ALDH activity and hepatic (C) acetaldehyde level. 간조직에서 acetaldahyde 는미토콘드리아에서 ALDH 의촉매작용에의해 acetic acid 로대사된다. 일반적으로동양인의 ADH 활성은서양인에비해우수한반면, ALDH 활성은서양인에비해 40-50% 낮은것으로알려져있다 (Wall TL 등 1997). ALDH 활성은알코올섭취및알코올성간질환이나간손상환자등간조직이손상될경우에감소되는것으로나타났다 (Vidal F 등 1998, Noh KH 등 2009). 동물이나임상실험에서도고위험수준의알코올이나만성음주에따른 ALDH 의활성저하로 acetaldehyde 나알코올유해대사산물이축적되어알코올성간손상이유발된다는여러보고들이있다 (Jo YO 등 2007, Jung HK 등 2008, Lee HI 등 2011, Kim MK 등 2013). 또한이와같은연구들 (Vidal F 등 1998, Jo YO 등 2007, Jung HK 등 2008, Noh KH 등 2009, Lee HI 등 2011, Kim MK 등 2013) 에서는하향주, 민들레즙, 헛개나무추출물을함유한오이발효음료, 차나무추출물및저분자키톤산함유해양심층수의공급이 ALDH 활성을증가시켜숙취해소에도움을줄수있었다고밝혔다. 그러나이들의연구결과와는상반되게 Park SH 등 (2004) 은체중 kg 당 5.5 g 의알코올을경구투여한후 2 시간이지난다음간조직의 ALDH 활성을측정하였을때정상군에비해알코올투여군에서 76.0% 의유의적으로활성이증가하였고, 보이차추출물을급여하자 ALDH 활성저하와함께 87.26% 의혈중의알코올농도의감소효과를보였다고하였다. 그러나본연구에서는 Fig. 9 와같이혈청중의알코올농도 (r=0.594, p<0.01) 및혈청 (r=0.477, p<0.01) ; 32(6):

9 826 박순자등 Korean J Food Cook Sci 과간조직 (r=0.491, p<0.05) 중의 acetaldahyde 농도는 ALDH 활성과양의상관관계를보였다. 그결과혈청에서는세수준의천마분말이, 간조직에서는 100 mg 과 300 mg 의천마분말의급여로 ALDH 의활성조절과함께알코올과 acetaldhyde 농도의감소효과가나타났다. Ⅳ. 요약및결론 본연구는동결건조시킨천마분말이간기능과알코올대사에미치는영향을동물실험을통해분석하여알코올성간손상에대한건강기능성식품의소재로서천마의활용방안을알아보고자실시하였다. 실험동물은정상대조군 (EC), 알코올대조군 (EC), 알코올과체중 kg 당천마분말 100 mg(eg100), 200 mg(eg200), 300 mg 투여군 (EG300) 으로나누었으며, 알코올은체중 kg 당 3 g 을투여하였다. 알코올과천마분말을 3 일동안매일 1 회씩일정시각에급여한다음혈청과간조직에서 GOT 와 GPT 활성, 알코올분해물질및효소에대해측정하였다. 그결과, 알코올의투여로혈청내의 GOT 의활성은유의미하지는않지만 100 mg 의천마분말의투여로낮출수있었으며, GOT 활성은혈청내알코올농도와양의상관관계 (r=0.531, p<0.01) 를보였다. 100 mg 의천마분말을급여하였을때 ADH 효소의활성유도로혈청내의알코올농도의저하효과를얻을수있었다 (p<0.05). 혈청중의알코올 (r=0.633, p<0.01) 과 acetaldahyde 농도 (r=0.477, p<0.01) 및간조직 (r= p<0.05) 중의 acetaldahyde 농도는 ALDH 활성과모두양의상관관계를보였다. 그러나세수준의천마분말의급여로혈청중 ALDH 의활성변화와함께알코올과 acetaldehydy 농도의감소효과를얻었으며, 간조직에서는천마분말의투여량에의존적인 acetaldehyde 농도의감소효과를얻었다. 특히, 알코올투여군에게 100 mg 의천마를급여했을때 ADH 의활성을유도하여알코올의분해를다소증가시켰다는측면에서알코올의대사를촉진하고간기능보호를위한기능성식품으로써천마가효과적인것으로판단된다. 그러나알코올과천마와의상호관련성에대한세밀한연구를통하여음주시적절한천마복용량에대한가이드라인이필요할것으로보여진다. Conflict of Interest No potential conflict of interest relevant to this article was reported. References Choi GH, Kim JG, Kwon ST Protective effects of food including Hovenia dulcis on acute alcohol intoxication. J Korean Soc Food Sci Nutr 40(8): Chung YI, Bae IY, Lee JY, Chun HS, Lee HG Protective effects of branched-chain amino acid (BCAA)-enriched corn gluten hydrolysates on ethanol-induced hepatic injury in rats. Korean J Food Sci Technol 41(6): Das SK, Vasudevan DM Alcohol-induced oxidative stress. Life Sci 81(3): Gill K, Amit Z, Smith BR The regulation of alcohol consumption in rats: The role of alcohol-metabolizing enzymes-catalase and aldehyde dehydrogenase. Alcohol 13(4): Han CK, Seong KS, Lee KW, Park SS, Jeong JY, Kim SS Effects of Omija (Schizandra chinesis Baillon) concentrate on blood alcohol clearance and hepatoprotective function in rats induced by acute ethanol intoxication and chronic ethanol treatment. J Korean Soc Food Sci Nutr 43(8): Hong HD, Shim EJ, Kim KI, Choi SY, Han CK Effect of Gastrodiae elata blume components on systolic blood pressure and serum lipid concentrations in spontaneously hypertensive rats fed high fat diet. J Korean Soc Food Sci Nutr 36(2): Hwang SJ, Choi HM, Park HJ, Lee JS, Heo D, Kim MR Effects of medicinal herbal drink on alcohol metabolic enzyme in drunken rats. Korean J Orient Physiol Pathol 24(4): Jeon TW, Lee ES, Lee YS, Han OK, Bae JC, Kim KJ, Kim HJ Eliminatory effect of mixed extract of Hovenia Dulcis Thunb and Gastrodia Elata on ethanol-induced hangover in rats. Korean J Orient Physiol Pathol 16(5): Jo YO, Koo SJ, Choi IS, Kong YH, Choi SY Effects of Camellia sinesis extracts on the antioxidant system and alcohol down-regulation enzymes in sub-acute ethanol treated ICR mice. J Korean Soc Food Sci Nutr 36(9): Jung BS Metabolic effects of alcohol. Korean J Food Nutr 4(2): Jung HK, Park CD, Hwang MH, Park SC, Kim DI, Hong JH Characteristics of alcohol metabolism of Hahyangju in rats. Korean J Food Preserv 15(1): Keshavarzian A, Fields JZ, Vaeth J, Holmes EW The differing effects of acute and chronic alcohol on gastric and intestinal permeability. Amer J Gastroenterol 89(12): Kim HJ, Kwon OJ, Lee AR, Roh SS, Seo YB Protective effects of Gastrodia rhizoma and steamed & fermented Gastrodiae rhizoma with anti-oxidant efficacy and suppression of NFkB signaling pathway on LPS-induced liver injury. J Appl Biol Chem 59(3): Kim IH Pathophysiology of alcoholic liver disease. Clin 2016; 32(6):

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