Journal of Radiation Industry 7 (1) : 75~8 (213) Review Paper 방사선유도돌연변이체, 블랙베리발효음료의알코올대사및숙취억제효과 조병옥 소양강 이창욱 조정근 우현심 진창현 정일윤 한국원자력연구원첨단방사선연구소 Effect of Fermented Blackberry Drinks Formed from Radiation-induced Mutant on Metabolism and Hangover in Rats Byoung Ok Cho, Yangkang So, Chang Wook Lee, Jung Keun Cho, Hyun Sim Woo, Chang Hyun Jin and Il Yun Jeong Advanced Radiation Technology Institute, Korea Atomic Energy Research Institute, Jeongeup 58-185, Korea Abstract - This study was designed to elucidate the effect of fermented blackberry drinks () on alcohol metabolism and hangover in alcohol-treated rats. We showed that the administration of increased the activity of alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH) in alcohol-treated rats. Moreover, the administration of reduced the serum alcohol and acetaldehyde concentrations in alcohol-treated rats. Furthermore, the administration of attenuated the levels of serum aspartate aminotransferase and alanine aminotransferase in alcohol-treated rats. Taken together, these results suggest that plays an important role in alcohol metabolism and liver function by reducing blood alcohol and acetaldehyde through the activation of ADH and ALDH in alcohol-treated rats and could be used as a functional anti-hangover drinks. Key words : Fermented blackberry drinks,, dehydrogenase, Aldehyde dehydrogenase 서 현대사회가빠른경제성장과생활수준의향상에도불구하고매년음주인구및알코올소비량이증가하고있다. 이로인하여알코올의과량또는만성적인섭취가 론 Corresponding author: Il Yun Jeong, Tel. +82-63-57-3, Fax. +82-63-57-3159, E-mail. iyjeong@kaeri.re.kr 늘어나면서알코올성간질환환자가증가하고알코올성간질환사망률또한높아지면서인체에대한유해성논란이증폭되고있는추세이다 (Kim et al. 5; Kim et al. 212). 알코올을섭취하였을경우흡수된알코올은체내에서영양소를과잉요구하거나흡수장애를일으키는원인이된다. 또한체내에서알코올을대사하는과정에서간에존재하는 alcohol dehydrogenase (ADH) 효소가활성화되어산화가일어나반응성이좋은아세트알데히드 75
76 조병옥 소양강 이창욱 조정근 우현심 진창현 정일윤 (acetaldehyde) 가증가하게되고, 아세트알데히드는 aldehyde dehydrogenase (ALDH) 효소에의해 acetate로산화가일어나게된다 (Han et al. 7; Chung et al. 9). 대사중생성되는아세트알데히드가분해가되면숙취가해소되지만, 아세트알데히드와자유라디칼의형성및알코올의산화로인한환원상태로의변화가숙취유발을비롯하여간에지방형성을촉진시켜간조직의손상이일어나게되며이는알코올성간질환으로이어지게된다. 이러한산화적손상으로부터간의건강을보호하고복구하고자체내에서는항산화효소들의분비가증가함을확인하였다 (Han 213). 이와함께항산화물질들이산화적손상을완화시키는것으로보고되었으며, 특히나 flavonoid와같은 polyphenol 물질을섭취할경우간조직의항산화능력을높여서손상을억제시킨다는보고가있다 (Cha 7). 최근들어, 항산화물질을천연물에서얻고자하는관심이증가하면서오래전부터약용또는식용으로사용되어온베리류의항산화활성에대한연구가활발하게이루어지고있다. 베리류는나무딸기류라고도칭하며장미과 (Rosaceae) 에속하여있는관목성식물이다 (Shin et al. 8; Sung 212). 연구에의하면나무딸기류에는 anthocyanin, flavonoid 그리고 polyphenol 등의다양한생리활성물질이포함되어있으며열매색이진할수록천연색소의함량이높아항산화효능이높다고알려져있다. 이외에도항암, 항염증, 순환기장애억제, 또는통증완화등에효과가있다고보고되어있다 (Oh et al. 8; Jung et al. 212; Han 213). 또한본연구진은방사선돌연변이신품종인블랙베리메이플의성분분석과항산화효능을확인한바있다 (Cho et al. 211). 그렇지만블랙베리의알코올유도에의한간손상보호에관한연구자료는매우부족한실정이다. 그리하여본연구에서는방사선돌연변이육종기술로얻어진신품종블랙베리메이플로제조된블랙베리발효음료가알코올을투여한쥐의체내알코올대사와숙취에미치는효과를확인하여간기능보호와숙취해소음료개발을위한기초자료를제공하고자하였다. 재료및방법 1. 블랙베리발효음료의제조본실험에서사용한블랙베리 (Rubus fruticosus L.) 는방사선돌연변이육종기술로얻어진신품종블랙베리메이플 (Ryu et al. 212) 을 211년전북완주에서재배하여생산된것을구입하여알코올및초산발효를통하여식 초발효원액을제조한후식이섬유, 벌꿀, 사과농축액, 구연산, 비타민정제수등을첨가하여블랙베리발효음료를제조하여실험에사용하였다. 2. 실험동물및처치실험동물은생후 6주령의수컷 Sprague-Dawley (18~ g) 계흰쥐 24마리를오리엔트바이오 ( 서울, 한국 ) 에서구입하였다. 항온항습 ( 온도 23±2 C, 습도 ±5%) 및 12시간간격의명암주기조건하에서 1주간고형식이와물로적응시킨후난괴법에의하여대조군, 알코올처리군, 알코올과블랙베리발효음료 () 1일처리군, 알코올과블랙베리발효음료 () 6일처리군으로각군당 6 마리씩나누었다. 실험에사용한알코올은전북전주에위치한주정판매월드주식회사로부터발효주정알코올을구입하여사용하였다. 알코올과블랙베리발효음료 () 6일처리군은블랙베리발효음료를 6 ml kg -1 으로 6일간매일경구투여하였다. 알코올과블랙베리발효음료 () 1일처리군은블랙베리발효음료를 6 ml kg -1 으로알코올투여1시간전에 1회경구투여하였다. 대조군과알코올처리군은식염수를 6 ml kg -1 으로 6일간매일경구투여하였다. 블랙베리발효음료마지막투여 1시간후에알코올 3g kg -1 를경구투여하였다. 알코올투여 1시간후에미정맥을통해채혈하고, 5시간후후대정맥을통해채혈한후희생하였다. 채혈한혈액은 3분간실온에방치한후 rpm, 4 C 에서 15분간원심분리하여혈청을분리하여실험에사용하였다. 3. 혈청중알코올대사효소활성측정혈청중의 alcohol dehydrogenase (ADH) 와 aldehyde dehydrogenase (ALDH) 효소활성은 Biovision (Milpitas, CA, USA) 사로부터구입한 alcohol dehydrogenase activity assay kit와 aldehyde dehydrogenase activity assay kit를이용하여제조사에서제시한방법에따라분리된혈청으로부터측정하였다. 4. 혈중알코올농도및아세트알데히드농도측정혈액중의알코올과아세트알데히드농도는 Megazyme (Wicklow, Ireland) 사의에탄올측정용 kit와아세트알데히드측정용 kit를이용하여제조사에서제시한방법에따라혈청으로부터측정하였다. 5. 혈청중간기능지표효소 AST와 ALT 측정분리된혈청으로부터간기능지표효소인 AST와 ALT
블랙베리발효음료의알코올대사및숙취억제효과 77 효소활성은 clinical biochemistry automatic analyzer (Hitachi model 718, Japan) 를이용하여측정하였다. 3 3 # ## 6. 통계처리모든실험결과는실험군당평균 ± 표준편차로나타내었고, 각군간의통계적유의성에대한검증은 student s t-test를이용하여, p.5일때통계적으로유의성이있다고판단하였다. (% control) 결과및논의 1. 블랙베리발효음료투여시혈중알코올과아세트알데히드농도에미치는영향숙취의주원인물질은알코올의일차대사산물인아세트알데히드로알려져있다. 이에본연구에서는블랙베리발효음료섭취가알코올투여된 rat의숙취에미치는영향을확인하기위하여혈중알코올과아세트알데히드농도를측정하였다. Fig. 1에서나타낸바와같이, rat에알코올을투여하였을경우 1시간후부터 5시간후까지혈중알코올과아세트알데히드농도가유의적으로증가하였지만, 블랙베리발효음료를 1일간 1회또는6일간 6회투여하였을경우 1시간후부터 5시간후까지혈중알코올과아세트알데히드농도가유의적으로감소하는것을확인하였다. 본연구결과는헛개나무열매추출물을함유한건강음료가알코올을투여한 rat의혈중알코올과아세트알데히드농도를감소시켜숙취해소효능을나타낸다는보고와유사한결과이다 (Park et al. 6). 또한, Kim et al. (211) 은복합약용식물추출물이알코올을투여한 rat의혈중알코올과아세트알데히드농도를감소시킨다고보고하였다. 따라서블랙베리발효음료를투여할경우혈중알코올과아세트알데히드농도를감소시킴으로써숙취해소효능을나타내는것으로사료된다. 2. 블랙베리발효음료투여시알코올대사효소활성에미치는영향알코올은주로간에서 ADH, cytochrome P4 2E1 및 catalase에의해숙취의주된원인물질인아세트알데히드가되고, 아세트알데히드는 ALDH에의해아세트산 (acetic acid) 으로산화된다. 아세트산은구연산회로를통해에너지를생성하고최종적으로뇨와 CO 2 로배설된다 (Lee et al. 211a). 따라서블랙베리발효음료섭취가 Acetaldehyde (% control) # 알코올투여된 rat의알코올대사에미치는영향을확인하기위하여알코올대사효소인 ADH와 ALDH 효소활성을측정하여 Fig. 2에나타내었다. Rat에알코올을투여하였을경우 1시간후 ADH와 ALDH 효소활성은유의적인차이를보이지않았으며, 5시간후 ADH 효소활성은증가하였으나 ALDH 효소활성은증가하지않은반 면, 블랙베리발효음료를 1 일간 1 회투여시 ADH 와 ALDH 의효소활성이소폭증가하였으며, 6일간 6회투여하였을경우 1시간후부터 5시간후까지 ADH와 ALDH의효소활성이유의적으로증가하는것을확인하였다. 비 Fig. 1. Effect of on serum ethanol and acetaldehyde concentrations in alcohol-treated rats. Blood samples were prepared from rats and tested for alcohol and acetaldehyde concentrations. Results are expressed as mean±sd (n=6). # p.1 vs. control 1 h, p.5 vs. alcohol 1 h, p.1 vs. alcohol 1 h, ## p.1 vs. control 5 h, p.1 vs. alcohol 5 h.
78 조병옥 소양강 이창욱 조정근 우현심 진창현 정일윤 3 3 ADH activity (% control) ALDH activity (% control) 18 16 14 12 8 6 4 ALT (IU l -1 ) AST (IU l -1 ) 8 7 6 4 3 2 2 1 Fig. 2. Effect of on serum ADH and ALDH activities in alcohol-treated rats. Blood samples were prepared from rats and tested for ADH and ALDH activities. Results are expressed as mean±sd (n=6). p.5 vs. alcohol. Fig. 3. Effect of on serum AST and ALT levels in alcoholtreated rats. Blood samples were prepared from rats and tested for AST and ALT levels. Results are expressed as mean±sd (n=6). 록단회투여시 ALDH의효소활성이유의적인영향을나타내지않았지만, 본실험결과들은알코올을투여한 rat에서약용식물추출물이첨가된미네랄이 ADH와 ALDH의효소활성을증가시킨다는보고와유사한결과이다 (Lee et al. 211b). 따라서블랙베리발효음료의경우 ADH와 ALDH의효소활성을증가하여숙취해소효능을나타내는것으로판단된다. 3. 혈청중 AST 및 ALT 활성변화 Rat에알코올투여시간손상지표효소인 AST와 ALT 효소가간으로부터혈액으로방출되어지며, 알코올성간손상에서중요한인자로알려져있다 (Ko et al. 6; Choi et al. 211). 이에본연구에서는알코올에의한간손상에서블랙베리발효음료의간기능보호효능을확인하기위하여혈청 AST와 ALT 활성을측정하였다. Fig. 3 에서나타난바와같이, rat에알코올을투여하였을경우혈액내에 AST와 ALT levels이증가하였지만블랙베리발효음료를 1일간 1회또는6일간 6회투여할경우혈중 AST와 ALT 효소활성이감소하는것을확인하였다. 따라서블랙베리발효음료섭취시알코올성간손상에보호활성을나타내는것으로추정할수있다.
블랙베리발효음료의알코올대사및숙취억제효과 79 결 본연구에서는기존에항산화및항암효능이우수한것으로알려진블랙베리추출물을주성분으로하여제조된블랙베리발효음료가알코올을투여한쥐의혈중알코올농도와체내알코올대사에어떠한영향을미치는지확인하여간기능보호와숙취해소음료개발을위한기초자료를제공하고자실시하였다. 알코올을투여한 rat에서는혈중알코올과아세트알데히드농도가증가하였고, ADH와 ALDH 효소활성은유의적인차이를보이지않았으며, AST 및 ALT 활성이소폭증가하였지만, 블랙베리발효음료를경구투여한 rat에서는혈중알코올과아세트알데히드농도가감소하였고, ADH와 ALDH 효소활성은증가하였으며, AST 및 ALT 활성이감소하는경향을나타내었다. 따라서블랙베리발효음료를투여할경우 ADH와 ALDH의효소활성을증가하여혈중알코올과아세트알데히드농도를감소시킴으로써체내알코올대사에효과적인효능을나타내는것으로판단되며, 간기능보호효과를가진기능성숙취해소음료로사용될수있을것으로사료된다. 사 본연구는미래창조과학부지원방사선기술개발사업에의하여수행되었으며이에감사드립니다. 론 사 참고문헌 Cha HS, Youn AR, Park PJ, Choi HR and Kim BS. 7. Comparison of physiological activities of Rubus coreanus Miquel during aaturation. J. Korean Soc. Food Sci. Nutr. 36:683-688. Cho BO, Ryu HW, Jin CH, Choi DS, Kang SY, Kim DS, Byun MW and Jeong IY. 211. Blackberry extract attenuates oxidative stress through up-regulation of Nrf2-dependent antioxidant enzymes in carbon tetrachloride-treated rats. J. Agric. Food Chem. 59:11442-11448. Choi GH, Kim JG and Kwon ST. 211 Protective effect of food including Hovenia dulcis on acute alcohol intoxication. J. Korean Soc. Food Sci. Nutr. 4:117-1112. Chung YI, Bae IY, Lee JY, Chun HS and Lee HG. 9. Protective effects of branched-chain amino acid (BCAA)-enriched corn gluten hydrolysates on ethanol-induced hepatic injury in rats. Korean J. Food Sci. Technol. 41:76-711. Han DO, Park SJ, Sur BJ, Lee HJ, Kim SH and Hahm DH. 7. Protective effect of the herbal combination HCE-2 on alcohol metabolism and alcohol-induced liver injury in ethanolloaded rats. Korean J. Oriental Physiology and Pathology 21:181-186. Han SZ. 213. Protective efficacies of Aronia melanocarpa (Blackberry) on the allyl alcohol-damaged hepatocyte of mice. Kor. J. Pharmacogn. 44:91-96. Jung H, Lee HJ, Cho H and Hwang KT. 212. Antioxidant and anti-proliferative activities of Rubus fruits in Korea. J. Korean Soc. Food Sci. Nutr. 41:1649-1655. Kim DH, Sung NY, Kim JS, Kim JH, Hwang YJ and Lee JW. 211. metabolizing and antioxidant activities of complex herbal extracts from medicinal plants. Food Sci. Biotechnol. 2:1337-1345. Kim J, Nam KS and Noh SK. 212. Cherry silverberry (Elaeagnus multiflora) wine mitigates the development of alcoholic fatty liver in rats. J. Korean Soc. Food Sci. Nutr. 41: 57-64. Kim J, Seo B, Choi S and Choi H. 5. Effects of water extracts from Haninjinho-tang on hyperlipidemia and liver damage induced by alcohol. Kor. J. Herbology 2:9-17. Ko BS, Jang JS, Hong SM, Kim DW, Sung SR, Park HR, Lee JE, Jeon WK and Park S. 6. Effect of new remedies mainly comprised of Hovenia dulcis Thunb on alcohol degradation and liver protection in Sprague Dawley male rats. J. Korean Soc. Food Sci. Nutr. 35:828-834. Lee HI, Seo KI, Lee JS, Hong SM, Lee JH, Kim MJ and Lee MK. 211a. Effect of fermented cucumber beverage on ethanol metabolism and antioxidant activity in ethanoltreated rats. J. Korean Soc. Food Sci. Nutr. 4:199-116. Lee SJ, Kim Andre, Lee JH, Kim MH, Lee BS, Jee YT, Bin JH and Ha JM. 211b. Effects of minerals added to medicinal plant extracts on alcohol-induced oxidative stress and alcohol metabolism in rats. J. Korean Soc. Food Sci. Nutr. 4: 393-4. Oh HH, Hwang KT, Kim M, Lee HK and Kim SZ. 8. Chemical characteristics of raspberry and blackberry fruits produced in Korea. J. Korean Soc. Food Sci. Nutr. 37:738-743. Park EM, Ye EJ, Kim SJ, Choi HI and Bae MJ. 6. Eliminatory effect of health drink containing Hovenia dulcis Thunb extract on ethanol-induced hangover in rats. Korean J. Food Culture 21:71-75. Ryu J, Kim DS, Ha BK, Kim JB, Kim SH, Jeong IY, Jo HJ, Kim EY and Kang SY. 212. Growth characteristics and morphological variation analysis of mutant lines derived from gamma-ray and chemical mutagen treatments in Rubus fructicosus L. J. Radi. Indus. 5:347-352. Shin JS, Lee JC, Sim OK, Yoon TY, Cho HJ and Kim EY. 8. Plant Regeneration via Multiple Shoot Formation
8 조병옥 소양강 이창욱 조정근 우현심 진창현 정일윤 from Sucker Explant of Hybrid Blackberry (Rubus fruticosus L. R. parvifolius L.). Korean J. Plant Res. 21(2):111-116. Sung KC. 212. A study on the pharmaceutical & chemical characteristics and analysis of natural Bokbunja extract. J. Korean Oil Chemists Soc. 29:42-411. Manuscript Received: July 1, 213 Revised: August 1, 213 Revision Accepted: August 1, 213