JNH 김정희(K14-11).fm

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1 Journal of Nutrition and Health (J Nutr Health) 2015; 48(2): 123 ~ pissn / eissn Research Article 고지방식이로유도한비만흰쥐에서함초 (Salicornia herbacea L.) 분말의항비만효과탐색 * 김미정 전현영 김정희 서울여자대학교자연과학대학식품영양학과 Anti-obesity effect of Korean Hamcho (Salicornia herbacea L.) powder on high-fat diet-induced obese rats* Kim, Mi Joung Jun, Hyun Young Kim, Jung Hee Department of Food and Nutrition, College of Natural Sciences, Seoul Women's University, Seoul , Korea ABSTRACT Purpose: The objective of this study was to examine whether Hamcho freeze-dried powder and Hamcho freeze-dried nano powder have an anti-obese effect on high fat diet-induced obese rats. Methods: Sprague Dawley rats were divided into 6 different groups, each of which was bred for 8 weeks with a different experimental diet: normal diet group (NC), high fat diet group (HFC), high fat diet + 2% Salicornia herbacea L. freeze-dried powder group (2% HS), high fat diet + 5% Salicornia herbacea L. freeze-dried powder group (5%HS), high fat diet + 1% Salicornia herbacea L. freeze-dried nano powder group (1% HSN), high fat diet + 2% Salicornia herbacea L. freeze-dried nano powder group (2% HSN). Results: The 5% HS group showed a weight loss effect in body weight, liver and adipose tissue by reducing the amount of dietary intake and food efficiency ratio. In addition, the 5% HS group showed a significant reduction of serum leptin concentration, while having a beneficial effect on the improvement of lipid metabolism such as increase the serum concentration of adiponectin. However, Hamcho freeze-dried nano powder did not show a weight loss effect. Conclusion: Overall data indicated that Hamcho seemed to have anti-obesity effects. In particular, consumption of 5% Hamcho freeze-dried powder might have beneficial effects on body weight, serum adiponectin level, and lipid profiles since it led to reduced body weight and growth of adipose tissues by suppressing dietary intakes. However conduct of further research studies is needed for analysis of active components and biochemical action mechanism of Hamcho. KEY WORDS: Salicornia herbacea L., anti-obesity effect, lipid profile, leptin, adiponectin 서 비만인구는현대사회로들어오면서우리나라뿐만아니라전세계적으로급속하게증가되고있으며, 이러한비만인구의증가는여러가지만성질환을야기하고나아가사회적, 경제적인문제를발생시킨다. 이로인하여세계보건기구 (World Health Organization, WHO) 에서는 비만이란단순히건강을해치는위험요인이아닌치료가필요한질병 이라고선언하였다. 1 비만은섭취량에비해소모하는열량이적어발생되는 론 대사성질환으로단순히체중만증가하는것이아닌체내지방조직이증가되는상태를말하며, 식습관과운동부족뿐만아니라다양한환경적유전적인요인이작용되는복합증후군이다. 2,3 비만을개선하기위한여러가지치료방법으로운동요법, 식사요법, 약물요법및수술등다양한방법이있으나, 비만치료제의화학합성물질은높은비만억제효능을나타내는반면부작용역시많이나타나는것으로알려져있다. 4 따라서최근에는안전하고부작용의위험이적은천연식물소재에대한관심이점차높아지고있으며, 특히우리나라자생식물을이용하여부가가치를 Received: August 6, 2014 / Revised: September 4, 2014/ Accepted: February 27, 2015 *This study was supported by a R&D grant of Korea Institute of Planning & Evaluation for Technology in Food, Agriculture, Forestry & Fisheries (IPET ). To whom correspondence should be addressed. tel: , jheekim@swu.ac.kr 2015 The Korean Nutrition Society This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License ( which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

2 124 / 함초분말의항비만효과 높이고비만치료에효과를나타내는기능성소재를개발하기위한여러가지연구가요구되고있다. 천연물을이용한항비만소재로는지방의소화와흡수를저해하는물질로 flavonoids, 키토산등이있으며, 식욕억제에관여하는물질로식이섬유소, 이눌린, hydroxycitric acid (HCA) 등이보고되고있다. 5-7 이외에도고추의 capsaisin, 녹차의 catechin 성분은체중및체지방감소에효과를나타냈고, 홍삼은에너지대사를활발히하여체내저장된지방량을감소시키는것으로보고되었다. 8,9 또한홍차와녹차는위장관에서췌장 lipase와 α-amylase 등지방과탄수화물의소화효소활성을감소시키는데기인하는것으로보고되었다. 10 함초 (Salicornia herbacea L.) 는명아주과에속하는식물로생김새가퉁퉁하고마디마디가튀어나온풀과같다하여 퉁퉁마디 로도불리어지며, 나문재, 칠면초, 해홍나물등과함께우리나라갯벌의지역에자생하는대표적인염생식물이다. 11,12 함초는염류가높은염습지대에서자라므로내염성이강하고다량의 NaCl을함유하고있으나, 다른식품에비해 K, Ca, Mg, Fe 등다양한천연미네랄역시많이함유하고있다. 13 또한필수지방산및필수아미노산역시풍부하며 50~60% 정도의과량의식이섬유소가함유되어있어함초는예로부터몸안에쌓인독소및숙변을배출시키고위장병, 소화불량등의좋은약재로도이용되어왔다. 14,15 최근기능성식품으로서의함초에대한관심이점차높아지면서생리활성에대한여러가지연구가수행되어져항산화 16 및항당뇨병, 17,18 항고혈압, 19 항암, 20 지질대사개선 18,21,22 등에관한많은연구결과들이보고되고있다. 특히 3T3-L1 adipocytes를이용한선행연구에서는함초물추출물이지방세포합성과정에서분화를억제하는데영향을미쳐 leptin 및 resistin 생성을감소하였고, adiponectin 생성을증가시켜비만과당뇨병및심혈관계질환예방효과를가질가능성이있을것으로보고되었다. 23 건강에대한관심이높아지면서건강을향상시키는데도움이되는여러가지기능성식품에관한연구역시증가되고있다. 최근에는각각의기능성물질의흡수를도울수있는여러가지방법들이연구되고있는데, 나노화기술이역시그중한가지로들수있다. 나노물질이란미립자크기의작은형태로나누어지는분말물질을말하며, 이러한나노화과정은기존의마이크로물질에비해비표면적이증가하고수용성및흡수율을증가시키는특징이있다. 24,25 칼슘과지질의경우에도나노화를통해비표면적과수용성을증가시켜체내흡수율을높일수있다는연구결과가보고되고있다. 26,27 그러나나노화물질은작은입자크기 로인해물질의특성이변화될수있고, 일부세포실험을통한연구에서마크로단위의물질보다나노화물질의흡수성과독성이증가하여세포의사멸에영향을미치는것으로보고되고있다. 28 그러므로기능성소재식품을이용한나노기술의적용은보다안정성과독성에대한연구가필요하다. 따라서본연구에서는한국산함초 (Salicornia herbacea L.) 의항비만효과를알아보고자함초동결건조분말과, 미세한입자로비표면적과수용성을증가시킨동결건조나노화분말이고지방식이로비만을유도한흰쥐의체중증가억제, 지질농도개선및비만관련호르몬을변화시키는지알아보고자실험을실시하였다. 연구방법 동물사육및실험설계본연구는 160~180 g 정도의 6주령된 Sprague Dawley (SD) Rat 수컷 63마리를나라바이오텍에서공급받았으며, 실험기간중사육실실내온도는 23 ± 2 o C 로유지시켰으며, 채광은 7:00~19:00으로일정하게조절하였다. 처음일주일간일반고형식이를공급하여실험실환경에적응시킨후, 각군간의차이가없도록체중을측정하여난괴법에따라 normal diet 섭취군 (NC, n = 12), high fat diet 섭취군 (HFC, n = 11), high fat diet + 함초동결건조분말 2% 섭취군 (2% HS, n = 10), high fat diet + 함초동결건조분말 5% 섭취군 (5% HS, n = 10), high fat diet + 함초동결건조나노화분말 1% 섭취군 (1% HSN, n = 10), high fat diet + 함초동결건조나노화분말 2% 섭취군 (2% HSN, n = 10) 등총 6군으로나누어 8주간사육하였다. 동물실험의모든절차및방법은서울여자대학교동물실험윤리위원회의권고에따라실시하였다 (IACUC-A-2). 실험식이는 AIN-76을기초로하여본실험목적에맞도록수정하여결정하였다. HFC군은고지방식이로단백질급원및무기질과비타민급원의구성비는 NC군과동일하지만, 옥수수전분의함량은줄이고지방의급원으로옥수수기름외에돼지기름 (lard) 15% 와콜레스테롤 1% 의함량을추가적으로공급하여지방이전체열량의약 40% 가되도록제조하였다. 함초동결건조분말섭취군 (2% HS, 5% HS) 과함초동결건조나노화분말섭취군 (1% HSN, 2% HSN) 은 HFC군의고지방식이조성에함초동결건조분말을 2%, 5% 로함초동결건조나노화분말을 1%, 2% 로함량을달리하여첨가하였으며, 첨가한만큼의양을옥수수전분에서제하여제조하였다 (Table 1). 함초동결건조분말의함량은신체표면적기준화기법 29

3 Journal of Nutrition and Health (J Nutr Health) 2015; 48(2): 123 ~ 132 / 125 Table 1. Composition of experimental diets (%) Ingredient Group NC HFC 2% HS 5% HS 1% HNS 2% HNS Casein DL-methionine Corn starch Sucrose Cellulose Corn oil Lard AIN-mineral Mix 1) AIN-vitamin Mix 2) Choline chloride Cholesterol Sodium cholate Hamcho freeze-dried powder Hamcho freeze-dried nano powder NC: normal diet group, HFC: high fat diet group, 2% HS: high fat diet group with 2% Salicornia herbacea L. freeze dryed powder, 5% HS: high fat diet group with 5% Salicornia herbacea L. freeze dryed powder, 1% HSN: high fat diet group with 1% Salicornia herbacea L. freeze dryed nano powder, 2% HSN: high fat diet group with 2% Salicornia herbacea L. freeze dryed nano powder 1) Composition of AIN-76 salt mixture 2) Composition of AIN-76 vitamin mixture 에근거하여 2% 와 5% 로설정하여식이중에혼합하여공급하였다. 나노화처리를한식품성분은표면적의증가로흡수율이약 2배증가되고, 26,27 과량의섭취는독성작용 28 의영향을준다는여러연구결과를토대로동결건조분말의 1/ 2에해당하는양인 1% 와 2% 로사료에첨가하였다. 이를통해함초동결건조분말과나노화분말의섭취가항비만효과에차이를나타내는지알아보기위해실험을진행하였다. 본실험에사용된함초동결건조분말은전라남도신안군에서채취한함초를이용하여제조하였으며, 함초동결건조나노화분말은동결건조분말을입도 3 μm로초미립분말화하여사용하였다. 모든실험식이는지방의산패를방지하기위해 1 kg씩 Pellet 형태로조제하고밀봉하여냉장보관하였다가 1일 1회일정시간에각군마다일정량 (20 g/ 일 ) 을공급하였다. 식이효율 (Food Efficiency Ratio, FER) 은실험기간에성장한체중증가량을같은기간동안섭취한식이의양으로나누어다음과같이계산하였다. Body weight gain for experimental period (g) FER = Food intake for experimental period (g) 였다. 채취한혈액에서 4 o C 3,000 rpm으로 20분간원심분리하여혈청을분리하였다. 분리된혈청은 e-tube에분주한후액체질소를사용하여급속동결시켜 -80 o C에서보관하였다가사용하였다. leptin, adiponectin 분석에사용하였다. 혈청의중성지질, 총콜레스테롤, HDL-콜레스테롤의함량은 Salè 등의방법을수정한효소법 30 ( 중성지질 : BCS Triglyceride Kit, Bio Clinical System Co., Anyang, Korea, 총콜레스테롤 : BCS Total Cholesterol Kit, Bio Clinical System Co., HDL-콜레스테롤 : BCS HDL- Cholesterol Kit, Bio Clinical System Co.) 을이용하여측정하였다. 혈청 leptin과 adiponectin 농도는각각 leptin mouse/rat enzyme immunoassay (EIA) kit (SPI-bio, Bretonneux, France) 와 adiponectin rat EIA kit (ALPCO Diagnostics, Salem, NH, USA) 를사용하여 sandwich type enzyme linked immunosorbent assay (ELISA) 법으로측정하였으며, plate reader (Spectra Max 250, Molecular Devices, Sunnyvale, CA, USA) 를이용하여 450 nm에서측정하였다. 혈청지질, leptin 및 adiponectin 농도측정실험동물은 8주간의실험기간종료후희생전에 12시간동안절식시키고, diethyl ether로마취시켜혈액과간, 부고환지방조직을적출하였다. 적출한부고환지방조직 ( 좌, 우 ) 은차가운생리식염수로세척한후여과지에서수분과잔여물을제거하여좌우지방조직각각의중량을측정하 간의지질, 단백질및 FAS 농도측정간조직은혈액채취후즉시적출하여차가운생리식염수로세척하고여과지에서여분의수분과잔여물을제거한후무게를측정하였다. 간의 homogenate와 cytosol 분획은 Masmoudi (1989) 의방법 31 에따라분리하였다. 얻어진간세포질역시 e-tube에분주하고액체질소를사용하

4 126 / 함초분말의항비만효과 여급속동결시켜 -80 o C deep freezer에서보관하였다가분석시사용하였다. 간조직속의지질은 Bligh와 Dyer의방법 32 을이용하여간 homogenate에서추출하였다. 간지질은혈청지질분석과동일한방법 30 을이용하여측정하였다. 간 homogenate 및세포질의단백질함량은 Lowry 방법 33 으로측정하였다. 이때표준물질로는 bovine serum albumin을사용하여계산하였다. 측정된단백질의양은간속에들어있는중성지질, 총콜레스테롤및 FAS의함량계산에이용하였다. Fatty acid synthase (FAS) 농도는간의 membrane 분획으로얻어낸 cytosol을이용하여분석하였으며, commercial ELISA kit (Uscn Life Science, USA) 를사용하여 ELISA 법으로 plate reader (Spectra Max 250, Molecular Devices, Sunnyvale, CA, USA) 를이용하여측정하였다. Table 2. Effect of Salicornia herbacea L. powder on final body weight, food intake and food efficiency ratio 1) Final body weight (g) Food intake (g/day) Food efficiency ratio NC (12) ± b ± 0.33 a ± b HFC (11) ± a ± 0.64 c ± a 2% HS (10) ± a ± 1.04 c ± a 5% HS (10) ± c ± 1.15 d ± b 1% HSN (10) ± a ± 0.45 bc ± a 2% HSN (10) ± a ± 0.43 ab ± a (n): number of animals, NC: normal diet group, HFC: high fat diet group, 2% HS: high fat diet group with 2% Salicornia herbacea L. freeze dryed powder, 5% HS: high fat diet group with 5% Salicornia herbacea L. freeze dryed powder, 1% HSN: high fat diet group with 1% Salicornia herbacea L. freeze dryed nano powder, 2% HSN: high fat diet group with 2% Salicornia herbacea L. freeze dryed nano powder Means with the different superscripts are significantly different at p < 0.05 by Duncan's multiple range test. 분변채취및지질분석실험동물의분변은희생하기전즉 8주차에 metabolic cage에서 24시간동안의분변을받아내어수집하였다. 수집된분변은상압가열건조법으로건조한후믹서기로분쇄하였으며, 각각의시료를 conical tube (15 ml) 에넣고뚜껑을잘밀봉하여냉동고 (-18 o C) 에보관하였다. 분쇄한분변속의지질역시 Bligh와 Dyer의방법 32 을이용하여추출하였다. 분변의중성지질과총콜레스테롤정량분석을하기위해서, 분석직전에분쇄한분변에서추출한지질에 chloroform 1 ml를일정하게넣어잘녹인후 0.1 ml를취하여 N 2 gas로 chloroform을휘발시킨후 anhydrous ethanol 0.2 ml를첨가하여다시지질을녹여사용하였다. 24시간분변지질은혈청지질분석과동일한방법 30 을이용하여측정하였다. 통계처리실험동물 63마리에대한모든실험결과는 Statistical Analysis System (SAS version 8.0) 을이용하여통계처리하였다. 모든측정치는평균과표준편차로표시하였으며, 함초동결건조분말과함초동결건조나노화분말농도에따른효과의차이를알아보기위하여실험결과를분산분석 (ANOVA) 으로분석하여 α = 0.05 수준에서 Duncan's multiple range test에의해유의성을검증하였다. 결 체중, 식이섭취량및식이효율실험종료후최종체중, 식이섭취량및식이효율에대한 과 결과는 Table 2 에제시하였다. 고지방식이군 (HFC) 의최종체중은정상식이군 (NC) 에비해유의하게증가하여고지방식이로인한체중증가가유도되었음을나타냈으며 (p < 0.05). 5% 함초동결건조분말을섭취한 5% HS군 ( g) 은 HFC군 ( g) 에비해유의하게감소하였고, NC군 ( g) 보다도유의적으로낮았다 (p < 0.05). 8주간의체중증가정도는모든군에서매주체중이증가하는경향을나타내었으나 5% HS군의경우 4주차이후체중의변화가크게나타나지않았다 (Fig. 1). 식이섭취량은 2% HSN군을제외한모든고지방식이군에서 NC군에비하여유의하게감소하였으며 (p < 0.05), 5% HS군 (16.95 g) 에서가장낮아 HFC군보다도유의하게감소하였다 (p < 0.05). 식이효율은에너지밀도가높은고지방식이군 (HFC) 이 ± 0.017로정상식이군 (NC) 의 ± 0.014에비하여유의하게증가하였으며, 체중증가율이가장적었던 5% HS군의식이효율이다른고지방군에비하여유의하게낮았다. 간및부고환지방조직무게간및부고환지방조직의무게를살펴본결과 (Table 3) 고지방식이의영향으로 NC군에비하여고지방식이를섭취한모든군에서간무게가유의하게증가하였으나 5% HS군의간무게는 HFC군에비해유의하게감소하였다 (p < 0.05). 상대적간무게역시 NC군에비하여모든고지방식이섭취군에서유의하게높았고 (p < 0.05), 1% 함초동결건

5 Journal of Nutrition and Health (J Nutr Health) 2015; 48(2): 123 ~ 132 / 127 Fig. 1. Body weight change Table 3. Effect of Salicornia herbacea L. powder on liver weight, relative liver weight, epididymal white adipose tissue weight and relative epididymal white adipose tissue weight 1) Liver weight (g) Relative liver weight (g/100g BW) Epididymal white adipose tissue-left (g) Relative Epididymal white adipose tissueleft (g/100g BW) Epididymal white adipose tissueright (g) Relative Epididymal white adipose tissueright (g/100g BW) NC (12) ± 0.92 c 2.82 ± 0.31 c 4.48 ± 0.78 ab 1.03 ± 0.20 a 4.52 ± 0.66 a 1.04 ± 0.18 a HFC (11) ± 2.79 a 6.66 ± 0.58 a 4.32 ± 0.55 ab 0.91 ± 0.10 ab 4.45 ± 0.49 a 0.93 ± 0.08 ab 2% HS (10) ± 3.94 a 6.33 ± 0.51 ab 3.99 ± 0.47 b 0.85 ± 0.13 b 4.18 ± 0.50 a 0.89 ± 0.11 b 5% HS (10) ± 3.93 b 6.52 ± 0.50 ab 2.46 ± 0.39 c 0.63 ± 0.07 c 2.50 ± 0.51 b 0.64 ± 0.10 c 1% HSN (10) ± 2.40 a 6.13 ± 0.39 b 4.23 ± 0.92 ab 0.85 ± 0.18 b 4.42 ± 0.90 a 0.89 ± 0.18 b 2% HSN (10) ± 4.02 a 6.43 ± 0.67 ab 4.60 ± 0.38 a 0.94 ± 0.07 ab 4.77 ± 0.49 a 0.97 ± 0.10 ab (n): number of animals, NC: normal diet group, HFC: high fat diet group, 2% HS: high fat diet group with 2% Salicornia herbacea L. freeze dryed powder, 5% HS: high fat diet group with 5% Salicornia herbacea L. freeze dryed powder, 1% HSN: high fat diet group with 1% Salicornia herbacea L. freeze dryed nano powder, 2% HSN: high fat diet group with 2% Salicornia herbacea L. freeze dryed nano powder Means with the different superscripts are significantly different at p < 0.05 by Duncan's multiple range test. 조나노화분말을섭취한 1% HSN군이 HFC군과비교해유의하게낮았다 (p < 0.05). 부고환지방조직과상대적부고환지방조직의무게를살펴보면, 왼쪽 / 오른쪽모두 5% HS군에서 HFC군에비해유의하게크게감소하여 NC군보다도유의적으로낮게나타났다 (p < 0.05). 24시간분변량, 수분함량및분변지질함량각군별 24시간분변량, 분변중수분및지질함량에대한결과는각군당 10마리씩총 60마리의분변을수집하여측정하였으며 Table 4에제시하였다. 24시간분변량은각군간의유의한차이는없었으며, 상압가열건조법으로건조한분변의무게와수분함량역시군간의유의한차이가없었다. 분변중중성지질과총콜레스테롤농도모두 NC 군에 비해모든고지방식이섭취군에서유의하게높았으나 (p < 0.05) 함초분말섭취에따른차이는없었다. 혈청, 간조직의지질함량및 FAS 농도혈청중성지질농도는 NC군에비해모든고지방식이섭취군에서유의하게높았으며, 함초분말섭취에따른유의한차이는없었다. 혈청총콜레스테롤농도역시 2% HS 군을제외한모든고지방식이군에서 NC군에비해유의하게높았고 (p < 0.05), 2% HS군의혈청총콜레스테롤농도는 HFC군에비해감소하는경향을보였다. 혈청 HDL- Cholesterol 농도는 NC군에서가장높았고 5% HS군의경우 2% HS군과 1% HSN군에비해유의적으로증가하였다 (Table 5). 간조직속의중성지질농도는고지방식이의영향으로 NC군에비하여 5% HS군을제외한고지방식이를섭취한

6 128 / 함초분말의항비만효과 Table 4. Effect of Salicornia herbacea L. powder on feces weight, feces water and lipid contents 1) Feces (g) Feces/Food intake (g/g) Dry feces (g) Water content (%) Triglyceride (mg/g dried feces) Total cholesterol (mg/g dried feces) NC (10) 1.47 ± 0.77 NS ± b 1.11 ± 0.53 NS ± NS 5.41 ± 2.57 b ± 4.90 b HFC (10) 1.97 ± ± ab 1.28 ± ± ± a ± a 2% HS (10) 2.32 ± ± a 1.42 ± ± ± 8.11 a ± a 5% HS (10) 2.05 ± ± a 1.28 ± ± ± a ± a 1% HSN (10) 1.90 ± ± ab 1.23 ± ± ± 9.94 a ± a 2% HSN (10) 2.30 ± ± ab 1.48 ± ± ± 5.31 a ± a (n): number of animals, NC: normal diet group, HFC: high fat diet group, 2% HS: high fat diet group with 2% Salicornia herbacea L. freeze dryed powder, 5% HS: high fat diet group with 5% Salicornia herbacea L. freeze dryed powder, 1% HSN: high fat diet group with 1% Salicornia herbacea L. freeze dryed nano powder, 2% HSN: high fat diet group with 2% Salicornia herbacea L. freeze dryed nano powder Means with the different superscripts are significantly different at p < 0.05 by Duncan's multiple range test. Table 5. Effect of Salicornia herbacea L. powder on serum lipid profiles 1) Triglyceride (mg/dl) Total cholesterol (mg/dl) 모든군에서유의하게증가하였으나 (p < 0.05), 함초분말섭취에따른유의한차이는없었다. 그러나 5% HS군의중성지질은 HFC군에비해유의하지는않지만감소하는경향을보였다. 간의총콜레스테롤농도역시고지방식이를섭취한모든군에서 NC군에비해유의하게증가하였으며 (p < 0.05), 함초분말섭취에따른유의한차이는없었다. 간세포질중 FAS 농도의경우, NC군이고지방식이를섭취한모든군들에비해유의하게증가하였으며 (p < 0.05), 함초분말섭취에따른차이는없었다 (Table 6). HDL-cholesterol (mg/dl) NC (12) ± a ± c ± a HFC (11) ± b ± ab ± ab 2% HS (10) ± b ± bc ± b 5% HS (10) ± 7.74 b ± ab ± a 1% HSN (10) ± b ± ab ± 8.29 b 2% HSN (10) ± b ± a ± ab (n): number of animals, NC: normal diet group, HFC: high fat diet group, 2% HS: high fat diet group with 2% Salicornia herbacea L. freeze dryed powder, 5% HS: high fat diet group with 5% Salicornia herbacea L. freeze dryed powder, 1% HSN: high fat diet group with 1% Salicornia herbacea L. freeze dryed nano powder, 2% HSN: high fat diet group with 2% Salicornia herbacea L. freeze dryed nano powder Means with the different superscripts are significantly different at p < 0.05 by Duncan's multiple range test. 혈청 leptin 및 adiponectin 농도혈청 leptin 농도는고지방식이섭취에따른유의한차이는없었으나모든함초분말섭취군에서 HFC군에비해낮은경향을보였으며, 특히 5% HS군에서는 HFC군에비해유의하게낮았다 (p < 0.05). 혈청 adiponectin 농도의경우고지방식이섭취에의해유의하게감소하였으며 (p < 0.05), Table 6. Effect of Salicornia herbacea L. powder on liver triglyceride and total cholesterol contents and cytosolic fatty acid synthase level 1) Triglyceride (mg/mg homogenate protein) Total cholesterol (mg/mg homogenate protein) Fatty acid synthase (ng/ml) NC (12) 1.64 ± 0.63 c 0.15 ± 0.05 c ± 2.63 a HFC (11) 2.96 ± 1.07 ab 1.44 ± 0.44 ab 6.97 ± 1.97 b 2% HS (10) 3.09 ± 1.02 ab 1.85 ± 0.53 a 7.27 ± 2.02 b 5% HS (10) 2.28 ± 0.65 bc 1.38 ± 0.48 b 7.90 ± 1.25 b 1% HSN (10) 3.13 ± 0.76 a 1.75 ± 0.43 b 7.49 ± 1.87 b 2% HSN (10) 3.40 ± 1.03 a 1.81 ± 0.58 a 6.60 ± 1.14 b (n): number of animals, NC: normal diet group, HFC: high fat diet group, 2% HS: high fat diet group with 2% Salicornia herbacea L. freeze dryed powder, 5% HS: high fat diet group with 5% Salicornia herbacea L. freeze dryed powder, 1% HSN: high fat diet group with 1% Salicornia herbacea L. freeze dryed nano powder, 2% HSN: high fat diet group with 2% Salicornia herbacea L. freeze dryed nano powder Means with the different superscripts are significantly different at p < 0.05 by Duncan's multiple range test. Table 7. Effect of Salicornia herbacea L. powder on serum leptin and adiponectin 1) Leptin (pg/dl) Adiponectin (µg/ml) NC (12) 1, ± a ± a HFC (11) 1, ± ab ± bc 2% HS (10) ± bc ± bc 5% HS (10) ± c ± b 1% HSN (10) 1, ± b ± c 2% HSN (10) ± b ± bc (n): number of animals, NC: normal diet group, HFC: high fat diet group, 2% HS: high fat diet group with 2% Salicornia herbacea L. freeze dryed powder, 5% HS: high fat diet group with 5% Salicornia herbacea L. freeze dryed powder, 1% HSN: high fat diet group with 1% Salicornia herbacea L. freeze dryed nano powder, 2% HSN: high fat diet group with 2% Salicornia herbacea L. freeze dryed nano powder Means with the different superscripts are significantly different at p < 0.05 by Duncan's multiple range test.

7 Journal of Nutrition and Health (J Nutr Health) 2015; 48(2): 123 ~ 132 / 129 함초동결건조분말섭취에따라유의하지는않지만증가하는경향을보여 HFC군에비해 5% HS군에서약간높았다 (Table 7). 고찰 최근비만인구가급속하게증가함에따라비만을예방및개선할수있는안전한천연식물소재에대한관심이높아지고있으며, 항비만효능을나타내는기능성식물소재들을찾아내어이들의작용기전을밝히는연구역시요구되고있다. 3 따라서본연구에서는최근관심이높아지고있는한국산함초 (Salicornia herbacea L.) 의항비만효과및함초분말의나노화에따른효능의차이를알아보고자흰쥐에게함초동결건조분말과함초동결건조나노화분말을고지방식이에혼합공급함으로서실험을실시하였다. 함초동결건조나노화분말은함초동결건조분말에비해비표면적을증가시켜소장에서함초의흡수율을높이기위해동결건조분말을입도 3 μm로초미립분말화하였다. 본연구에서최종체중은고지방식이군 (HFC) 이정상식이군 (NC) 에비해유의적으로높아고지방식이로인한비만이유도되었음을관찰할수있었다. 또한, 최종체중은 5% 함초동결건조분말섭취에의해유의하게감소되었으며고지방식이대조군 (HFC군) 뿐만아니라정상식이군 (NC군) 보다도유의하게낮게나타났다. 식이섭취량은에너지밀도가높은고지방식이섭취에의해감소하였으며, 5% 함초분말섭취에의해더욱감소하여가장낮은식이섭취량을보였고, 식이효율역시 5% HS 군에서가장낮았다. 이러한결과로볼때, 5% 함초동결건조분말섭취는식이섭취량에영향을미쳐체중증가를억제하는항비만효과를보이는것으로사료된다. 그러나흡수율증가효과를기대한분말의나노화는체중감소효능에차이를나타내지않았다. 일반적으로함초는총식이섬유함량이매우높은것으로보고되고있으며, 34,35 섬유소를다량함유하고있는김 (31.4%), 미역 (37.8%), 다시마 (29.3%) 등해조류에비해약 2배높은것으로보고되고있다. 36 또한, 결과에제시하지는않았지만본연구에서사용한함초동결건조분말과함초동결건조나노화분말의식이섬유소함량을한국식품연구원에서분석한결과, 각각 100 g당 41.3 g과 40.1 g의식이섬유소를함유하고있어모두 40% 이상의식이섬유소를함유하고있는것으로조사되었다. 이처럼함초에다량함유된식이섬유는에너지원으로이용되지못하고쉽게포만감을느끼게함으로써음식의섭취를줄이고체중과식이효율의감소를가져와비만억제에긍 정적영향을미칠수있다. 그러나함초가염분을함유하고있으므로함초의첨가비율이점차증가하면짠맛의강도역시증가하게되어식이섭취량이감소되고이에따라체증증가가상대적으로적게일어날수있음도배제할수없다. 또한, 2% 함초동결건조분말섭취는식이섭취량이나체중에영향을나타내지않았고, 2% 동결건조분말군과동일한식이섬유소를함유한 2% HSN군에서는오히려식이섭취량이고지방대조군에비해유의하게증가하였으므로 5% 함초동결건조분말에의한항비만효과가식이섬유소섭취에의한것인지는그에대한기전연구가필요하다. 아울러분말의나노화역시, 5% 나노화분말섭취군에대한실험을실시하지않은관계로나노화를통한섬유소입자의미세화가장내섬유소의흡수력을향상시켜, 26,27 분해되지않고장내에서작용하는식이섬유로서의효능이감소된결과인지아니면식이섭취를감소시킬수있는식이섬유소공급량이식이중 5% 까지증가되어야하는지에대한보완연구가필요할것으로사료된다. 고지방식이섭취는간에중성지질및콜레스테롤등의축적을일으켜간중량을증가시킨다. 37 본실험결과에서도간무게는고지방식이의영향으로 NC군에비하여고지방식이를섭취한모든군에서유의하게증가하였으며, 이는고지방식이에의해간내에콜레스테롤및중성지질등이축적되어중량을증가시킨다는결과와일치하는것으로보여진다. 38 함초분말섭취에따라서는 5% HS군의간무게가 HFC군에비해유의하게낮았으나, 상대적인간무게는 HFC군과유의한차이가없었다. 이는 5% 함초동결건조분말섭취군의식이섭취량감소로인한체중감소때문에체중을고려한상대적인간무게에는유의한차이가나타나지않은것으로사료된다. 일반적으로비만이란단순한체중의증가가아닌지방조직에중성지방이축적되어발생하는체지방량의증가를말하며, 특히복부지방함량이증가할수록대사성질환위험역시증가한다고보고되고있다. 39 중성지방의저장고인부고환지방조직의무게를살펴보면왼쪽 / 오른쪽모두 5% HS군이 HFC군에비해유의하게낮은것을볼수있으며, NC군보다도유의적으로수치가낮았다. 또한체중을고려한상대적부고환지방조직의무게에서역시 5% HS군이 HFC군이나정상식이군에비해유의하게크게낮았는데이러한결과는고지방식이와식이섬유를동시에섭취한흰쥐의체지방량이저지방식이섭취군과비슷한수치를나타낸 Artissa 등 40 의연구결과와비교할때함초에함유된식이섬유섭취가체지방조직감소에영향을미쳤을가능성도배제할수없다. 그러나, 본연구결과 5% 함초동결건조분말의섭취는식이섭취량을유의하게크게

8 130 / 함초분말의항비만효과 감소시켰기때문에이로인한간및부고환지방조직에서의지방축적억제효과가더크게나타났을것으로사료된다. 체중을고려한상대적부고환지방조직의무게는정상식이군 (NC) 에서가장높게측정되었는데이는높은식이섭취율로인해증가된탄수화물의섭취가혈중중성지질을높여지방조직의합성을증가시킨것으로사료되나이에관한연구는좀더진행되어야할것이다. 41 Metabolc cage에서 24시간동안수집한분변량을식이섭취량으로보정하여분석한결과함초동결건조분말섭취에따른분변량의유의한차이는없었고, 분변에서추출한지질농도역시고지방식이섭취에의해서는유의하게증가하였으나함초분말섭취에따른차이는없었다. 또한, 혈중중성지방, 총콜레스테롤및 HDL-콜레스테롤농도역시함초섭취유무에따른유의한차이는나타나지않아함초동결건조분말섭취가장내지방의흡수나혈중지질농도에는영향을미치지않는것으로여겨진다. 일반적으로고지방식이급여로인한지방산공급의증가는혈중중성지방과총콜레스테롤농도를증가시키고 HDL-콜레스테롤농도를감소시켜심혈관및관상심혈관질환을야기한다. 42 본실험결과에서도혈청총콜레스테롤함량은 2% HS군을제외한모든고지방식이섭취군에서유의적으로증가하였으나혈중중성지질농도는 NC군에서고지방식이를섭취한모든군에비해유의적으로증가한것을볼수있었다. 이는정상식이를섭취한 NC군이고지방식이를섭취한다른군보다탄수화물의섭취가많아지방으로저장되는대사과정에서중성지질의합성을늘려혈중농도를높여나타내는결과로생각된다. 41 간은콜레스테롤합성의주요장기로지단백을구성하여순환계로분비함으로서혈액의콜레스테롤의농도를조절하는기능을하며, 간의콜레스테롤농도는순환기계질환의유발에주요한지표가되고있다. 43 또한, 고지방식이는간에지방을축적시켜지방간의원인이되는것으로알려져있으며, 37 본연구에서도간조직의중성지질과총콜레스테롤함량이고지방식이섭취에의해유의하게증가됨을관찰할수있었다. FAS는 acetyl-coa carboxylase (ACC) 와함께지방합성을조절해주는주요조절효소로조직특이적으로지방합성이활발한간이나지방조직에서많이검출되며, 영양섭취상태에따라활성도가민감하게반응하는것으로알려져있다. 44 본연구에서간세포질의 FAS 농도를측정한결과, FAS 농도가고지방식이섭취에의해유의하게감소하였는데, 이는같은열량의저지방고탄수화물식이가고지방저탄수화물식이보다지방산합성이증가되었다고한인체연구 45 결과와일치하며, NC 군이식이섭취량도다른군보다높았고상대적으로탄수 화물의섭취도많았기때문에이로인한해당과정중간대사물질증가가 FAS 농도를높이는데기인하였을것으로여겨진다. 46 그러나, 본연구결과, 간조직내중성지질, 콜레스테롤함량및 FAS 농도모두함초동결건조분말섭취에따른유의한차이는관찰할수없었다. Leptin은지방세포의비만유전자에의해생성되는호르몬으로비만의경우 leptin의분비량이증가하는것으로알려져있으며, 혈중 leptin 농도는체지방량을나타내는지표로비만연구에많이사용되고있다. 47 본실험에서혈중 leptin 농도는 NC군이모든군에비해가장높게측정되었는데, 이는높은식이섭취로탄수화물의섭취가증가되고이로인해혈중중성지질과, 지방조직의합성이증가되면서 leptin의분비량을증가시킨것으로사료된다. 혈청중 leptin 농도의경우 HFC군에비해모든함초섭취군에서유의적이진않았지만 leptin의수치가낮게나타나는경향이보였으며, 특히함초동결건조분말첨가농도에따라 leptin 역시의존적으로감소하는경향을보였다. 특히 5% HS군은 HFC군에비해 leptin 농도가유의적으로감소하였는데, 5% 동결건조함초의섭취는식이섭취량을감소시키고, 이로인한체중및체지방감소효과를야기하여혈중 leptin의농도를감소시키는효과가나타났다. 혈중 adiponectin은비만한사람에게서그발현양이감소하며, 내장지방이많을수록더높은음의상관관계를나타내고 adiponectin 농도감소는심혈관계질환의위험을증가시키는것으로보고되고있다. 48,49 혈청중 adiponectin 농도에대한결과는고지방식이의영향으로 NC군에비하여고지방식이를섭취한모든군에서농도가유의적으로감소하였다. 특히함초동결건조분말군의경우유의적인차이는없었지만농도의존적으로증가하는것을볼수있으며, 5% HS군의경우 HFC군에비해 adiponectin의농도가증가하는경향이나타났다. 이는 5% 의함초동결건조분말의섭취가식이섭취량을감소시켜지방의축적을막아 adiponectin의분비에영향을미쳤을것으로추정된다. 결론적으로고지방식이와함께섭취한 5% 함초동결건조분말 (5% HS) 은식이섭취량을줄여체중, 간및지방조직의무게감소효과를나타냈다. 또한 5% HS군은고지방식이군 (HFC) 에비해혈중 leptin 농도를유의적으로감소시켰으며, 혈중 adiponectin 농도를증가시키는경향을보이는등지질대사개선에긍정적인영향을미치는것으로나타났다. 이러한결과는함초동결건조분말에많이함유된식이섬유소와관련이있을가능성이있으나정확한원인에대해서는더많은연구가필요하다. 또한, 본연구는자유식이를통해식이섭취량의통제가정확히이루어지지않았기때문에함초동결건조분말의항비만원인물질

9 Journal of Nutrition and Health (J Nutr Health) 2015; 48(2): 123 ~ 132 / 131 및생화학적인작용기전에대한더심도있는연구가필요하며, 나아가인체대상의임상실험을통한직접적인연구도필요하다. 요약 본연구에서는한국산함초 (Salicornia herbacea L.) 인퉁퉁마디의항비만효과를알아보고자동물실험을실시하였다. 함초동결건조분말및함초동결건조나노화분말의항비만효과를살펴본결과는다음과같다. 1) 최종체중은 5% 함초동결건조분말을섭취한 5% HS 군이 HFC군에비해유의하게감소하였으며, NC군보다도유의적으로낮게나타났다. 식이섭취량은 NC군에비하여고지방식이섭취군 (HFC, 2% HS, 5% HS, 1% HSN) 에서유의적으로감소하였고, 5% 함초동결건조분말을섭취한 5% HS군은고지방식이군보다유의적으로식이섭취량이낮았다. 2) 간무게는고지방식이의영향으로 NC군에비하여모든고지방식이섭취군에서유의하게증가하였고, 5% 함초동결건조분말섭취군은 HFC군과비교해유의하게낮았다. 부고환지방조직의무게역시왼쪽 / 오른쪽모두 5% 함초동결건조분말섭취군에서 HFC군에비해유의하게낮았으며, NC군보다도유의적으로수치가낮았다. 3) 혈청중성지질농도는 NC군이모든고지방식이섭취군에비해유의하게높았으며, 혈청총콜레스테롤농도는 2% HS군을제외한모든고지방식이군에서 NC군에비해유의하게높았다. 그러나, 혈청중성지질, 총콜레스테롤, LDL-콜레스테롤및 HDL-Cholesterol 농도모두 HFC군과비교하여함초섭취에따른유의한차이가없었다. 4) 간조직의중성지질과총콜레스테롤농도는 NC군에비하여모든고지방식이섭취군에서유의적으로증가하였다 (p < 0.05). HFC군은함초섭취군과유의적인차이가나타나지않았으나, 5% 함초동결건조분말섭취군의중성지질과총콜레스테롤농도는 HFC군에비해감소하는경향을보였다. 5) 간세포질중 FAS 농도의경우 NC군이모든고지방식이섭취군에비해유의적으로증가하였으며, 함초분말섭취에따른유의한차이는없었다. 6) 혈청 leptin 농도는 HFC군에비해모든함초섭취군에서 leptin의수치가낮게나타나는경향이보였다. 특히 5% HS군은 HFC군에비해 leptin 농도가유의적으로감소하였다. 혈청 adiponectin 농도는 NC군이모든고지방식이섭취군에비해유의적으로높았으며, 5% 함초동결건조분말섭취군의경우 HFC군비해그농도가증가하는경향이 나타났으나유의적인차이는없었다. 이상의실험결과에서고지방식이와함께섭취한 5% 함초동결건조분말은식이섭취량을줄여체중, 간및지방조직의무게를감소하였고, 혈중 leptin 농도를유의적으로감소시켰으며, 혈중 adiponectin 농도를증가시키는경향을보이는등지질대사개선에긍정적인영향을나타냈다. 이러한결과는함초의식이조절및지질개선건강기능성소재개발가능성을제시한것으로생각된다. References 1. Solomon CG, Manson JE. Obesity and mortality: a review of the epidemiologic data. Am J Clin Nutr 1997; 66(4 Suppl): 1044S- 1050S. 2. Albu J, Allison D, Boozer CN, Heymsfield S, Kissileff H, Kretser A, Krumhar K, Leibel R, Nonas C, Pi-Sunyer X, VanItallie T, Wedral E. Obesity solutions: report of a meeting. Nutr Rev 1997; 55(5): Grundy SM. Multifactorial causation of obesity: implications for prevention. Am J Clin Nutr 1998; 67(3 Suppl): 563S-572S. 4. Reddy P, Chow MS. Focus on orlistat: a nonsystemic inhibitor of gastrointestinal lipase for weight reduction in the management of obesity. Formulary 1998; 33(10): Burns AA, Livingstone MB, Welch RW, Dunne A, Rowland IR. Dose-response effects of a novel fat emulsion (Olibra) on energy and macronutrient intakes up to 36 h post-consumption. Eur J Clin Nutr 2002; 56(4): Delzenne NM, Cani PD, Daubioul C, Neyrinck AM. Impact of inulin and oligofructose on gastrointestinal peptides. Br J Nutr 2005; 93 Suppl 1: S157-S Zacour AC, Silva ME, Cecon PR, Bambirra EA, Vieira EC. Effect of dietary chitin on cholesterol absorption and metabolism in rats. J Nutr Sci Vitaminol (Tokyo) 1992; 38(6): Oh SJ, Kim YS, Park CY, Kim SW, Yang IM, Kim JW, Choi YK, Paeng JR, Shin HD. Body fat decreasing mechanisms of red ginseng compound. J Korean Soc Study Obes 2000; 9(4): Rhee SJ, Kim KR, Kim HT, Hong JH. Effects of catechin on lipid composition and adipose tissue in obese rats fed high fat diet. J Korean Soc Food Sci Nutr 2007; 36(5): Jeon JR, Kim JY, Lee KM, Cho DH. Anti-obese effects of mixture contained pine needle, black tea and green tea extracts. J Korean Soc Appl Biol Chem 2005; 48(4): Lee TB. Illustrated flora in Korea. Seoul: Hyangmoonsa; Lee YN. An illustrated book of the Korean flora. Seoul: Kyohaksa; Kim CS, Song TG. Ecological studies on the halophyte communities at western and southern coasts in Korea (IV): the halophyte communities at the different salt marsh habitats. Korean J Ecol 1983; 6(3): Do JR, Kim EM, Koo JG, Jo KS. Dietary fiber contents of marine algae and extraction condition of the fiber. J Korean Fish Soc 1997; 30(2): Jo YC, Ahn JH, Chon SM, Lee KS, Bae TJ, Kang DS. Studies on pharmacological effects of Glasswort (Salicornia herbacea L.).

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