Journal of Nutrition and Health (J Nutr Health) 2014; 47(5): 313 ~ 320 http://dx.doi.org/10.4163/jnh.2014.47.5.313 pissn 2288-3886 / eissn 2288-3959 Research Article Jaceosidin 이 streptozotocin 으로유도된당뇨쥐의혈당강하및항산화능에미치는영향 * 박은교 1 권병목 2 정인경 3 김정현 3 중앙대학교대학원가정학과, 1 한국생명공학연구원의과학융합연구본부, 2 중앙대학교체육교육과 3 Hypoglycemic and antioxidant effects of jaceosidin in streptozotocin-induced diabetic mice* Park, Eunkyo 1 Kwon, Byoung-Mog 2 Jung, In-Kyung 3 Kim, Jung-Hyun 3 1 Dept. of Home Economics, Graduate School, Chung-Ang University, Seoul 156-756, Korea 2 Division of Biomedical Convergent, Korea Research Institute of Bioscience & Biotechnology, Daejeon 305-806, Korea 3 Dept. of Physical Education, Chung-Ang University, Seoul 156-756, Korea ABSTRACT Purpose: In this study, we investigated the effects of jaceosidin on blood glucose regulation in type 1 diabetic mice. Methods: C57BL/6 mice were divided into four groups; normal control (Normal), diabetes control (D-Control), diabetes lowjaceosidin (D-0.005%), and diabetes high-jaceosidin (D-0.02%). Type 1 diabetes was induced by streptozotocin and mice were then fed a diet containing jaceosidin for eight weeks. Fasting blood glucose, oral glucose tolerance test, insulin tolerance test, lipid peroxidation, and antioxidant enzyme activities were assessed. Results: Jaceosidin supplementation for eight weeks had no effect on body weight, organ weight, and blood lipid profiles. However, jaceosidin supplementation significantly lowered fasting blood glucose level and reduced insulin resistance. We also found that jaceosidin supplementation increased antioxidant capacity by enhancement of catalase and GSH-px activities. Conclusion: These results suggest that jaceosidin could be a therapeutic candidate to ameliorate hyperglycemia through increase of antioxidant enzyme activity. KEY WORDS: jaceosidin, diabetes mellitus, antioxidant, lipid peroxidation 서 우리나라국민의사망원인중 4위를차지하고있는당뇨병은유병률이 2001년 8.6% 에서 2010년 9.7% 로증가하는추세에있다. 1 또한, 전세계적으로도약 2억 8,500만명의당뇨병환자들이있으며, 2030년에는약 4억 3,800만으로증가할것으로예상되고있어이를예방하고치료하고자하는노력이지속적으로이루어지고있다. 1 당뇨병의종류로는자가면역성, 원인불명성베타세포파괴로인해인슐린분비가감소되어발병하는제1형당뇨병과인슐린저항성, 베타세포기능부전에의한인슐린결함 론 으로발생하는제2형당뇨병이있으며, 이중제1형당뇨병은인슐린의절대적결핍으로발병이급속하게진행되고여러합병증을유발한다고알려져있다. 2-4 또한, 당뇨병으로인한합병증은혈당이높게유지되면서 reactive oxygen species (ROS) 가과다하게생성되어발생하는것으로알려져있다. 5-8 정상적인세포의경우유리기의생성과항산화방어시스템의활성이균형을이루고있어조직이나세포에손상을주지않으나 9 당뇨병으로인해혈당이높게유지되면체내유리기가과다생성되고, 이를방어하는항산화시스템의기능이감소되어균형이깨지게되고, 이로인해산화적스트레스가과도하게발생하게된다. 10-12 과도한 Received: July 24, 2014 / Revised: August 8, 2014 / Accepted: August 27, 2014 *This work was supported by the National Research Foundation of Korea (NRF) grant funded by the Korean Government (NRF- 2013R1A1A2006028). To whom correspondence should be addressed. tel: +82-02-820-5378, e-mail: jjhkim@cau.ac.kr 2014 The Korean Nutrition Society This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
314 / Jaceosidin 에의한항당뇨효능 산화적스트레스는알도스환원효소와 hexosamine 경로의활성을증가시키고, 포도당자기산화반응을유발하여최종당화산물 (advanced glycation endproducts; AGEs) 의형성을증가시킬뿐아니라 5-8 세포내의지질과산화물의생성을증가시켜당뇨병증상을악화시키고여러합병증을유발하는것으로알려졌다. 13,14 따라서당뇨병에의해높아진혈당을조절함과동시에체내의항산화능을향상시키는것이당뇨병뿐아니라합병증의예방및치료에효과적인것으로보고되고있어최근에는항산화능을가진천연식물소재기능성물질을탐색하여당뇨병의치료와합병증예방등에이용하고자하는연구가활발히진행되고있다. 쑥은예로부터복통, 출혈, 두통, 간염, 류마티스, 황달, 빈혈, 고혈압, 신경통, 발열등의질환에사용되어왔다 15. 최근에는약쑥 (Artemisia princeps) 의기능성성분으로 eupatilin 과 jaceosidin이분석되었으며, 16 이중 jaceosidin은강력한항산화력과함께항암, 항염증작용을갖는것으로보고되고있다. 17-19 또한, 약쑥에함유된 flavone이제2형당뇨동물모델연구에서항산화력을증가시켜혈당과지질대사를개선하는효과가있는것으로보고되고있다. 20,21 따라서, 항산화력과항염증작용을하는것으로알려진 jaceosidin 은산화적스트레스가높은당뇨병환자의산화적스트레스를억제하고, 혈당을낮춤으로써당뇨증상을완화시키는데효과가있을것으로판단된다. 그러나, jaceosidin이제1형당뇨시혈당조절이나그기전에미치는영향에대한연구는전무하므로, 본연구에서는 jaceosidin 섭취가당뇨시당대사와항산화능에미치는영향을규명하고, 그기전을밝힘으로써당뇨병및당뇨합병증의예방 치료에효과적인천연식품소재로서 jaceosidin의기능성을밝히고자한다. 연구방법 실험동물사육및처리중앙실험동물에서 5주령된수컷 C57BL/J mice를구입하여일주일간안정화시킨후, 정상군 (Normal) 을제외한 mice에 5일간 streptozotocin (55 mg/kg body weight) 을 i.p, 주사하여당뇨를유도하였다. 48시간후 6시간절식을시킨상태에서꼬리정맥으로부터채혈하여혈당을측정 (Accu-CHEK, Roche, Germany) 하였으며혈당치가 200 mg/dl 이상인실험동물을 3그룹 ( 당뇨대조군 (D-Control), 0.005% jaceosidin 섭취군 (D-0.005%), 0.02% jaceosidin 섭취군 (D-0.02%)) 으로나누어각그룹에해당하는실험식이를공급하였다. 체중과식이섭취량은주 2회측정하였으며, 실험기간동안의총체중증가량을동일기간동안의총식이섭취량으로나누어식이효율 (FER) 을계산하였다. 동물은중앙대학교내청정동물연구센터에서사육되었으며, 동물실험실은온도 22±1 o C, 습도 50 ± 1% 를유지하였고 12시간간격으로점등과소등을반복하였다. 실험식이는 AIN-93 purified rodent diet base를기초로하였으며, jaceosidin은각각 0.005%, 0.02% 를첨가하여 pellet 형태로제조 (( 주 ) 유니페이스, Seoul, Korea) 한후 4 o C에서냉장보관하였다. 실험식이의조성은 Table 1과같다. 본동물실험은중앙대학교의동물윤리위원회에의해승인되었다 ( 승인번호 : 13-0036). 혈당및내당능측정실험식이를공급한후 1주일간격으로꼬리정맥으로부터채혈하여공복혈당을측정하여혈당변화를측정하였다. 경구당부하검사는실험사육 7주경에 6시간을절식시킨다음공복혈당을측정하였으며, 이후 glucose (2 mg/kg body weight) 를경구투여하고 15, 30, 60, 120분후에꼬리정맥으로부터채혈하여혈당을측정하였다. 인슐린내성검사인슐린내성검사는실험사육 8주경에 6시간절식시켜공복혈당을측정한뒤 insulin (0.75 U/kg body weight) 을 i.p. 주사하여 15, 30, 60, 120분후에혈당의변화를측정하였다. 채혈및장기적출희생하기전 12시간공복시킨실험동물을에테르로마취한후헤파린을첨가한주사기를이용하여심장에서혈액을채취하였다. 채취된혈액은원심분리 (1,000 rpm, 20 min) 하여혈장을분리하고, 분석전까지 -80 o C에냉동보관하였다. 해부후간, 신장, 비장, 지방조직은혈액을제거한후여과지로습기를제거하였고장기무게를측정한후액 Table 1. Composition of experimental diets (g/kg diet) Groups Normal D-Control D-0.005% D-0.02% Ingredients Corn Starch Casein Dextrin Sucrose Soybean oil Cellulose Mineral mix. 1) Vitamin mix. 2) L-cysteine Choline bitartrate 397.4 200.0 132.0 100.0 70.0 50.0 35.0 10.0 3.0 2.0 397.4 200.0 132.0 100.0 70.0 50.0 35.0 10.0 3.0 2.0 397.3 200.0 132.0 100.0 70.0 50.0 35.0 10.0 3.0 2.0 397.2 200.0 132.0 100.0 70.0 50.0 35.0 10.0 3.0 2.0 Jaceosidin - - 0.05 0.2 1) AIN-93 mineral mixture 2) AIN-93 vitamin mixture
Journal of Nutrition and Health (J Nutr Health) 2014; 47(5): 313 ~ 320 / 315 체질소에냉동시켜 -80 o C 에냉동보관하였다. 간기능손상정도측정간기능지표인 glutamic oxaloacetic transaminase (GOT) 와 glutamic pyruvic transaminase (GPT) 의활성은 kit (Asan Chemical, Seoul, Korea) 을이용하여제시된실험방법에따라측정하였다. 간내지질과산화물함량측정간조직에서의지질과산화물 (thiobarbituric acid reactive substance, TBARS) 농도는 TBA 시약을이용하여측정한다. 표준용액은 1,1,2,2,-tetraethoxypropane을 methanol과증류수로희석하여농도별로만들었으며, 시료는간조직 0.025 g을 250 μl의 RIPA buffer에넣고 sonication하여준비하였다. 표준용액혹은간균질액에 1% 인산용액과 TBA 용액을혼합한후 60 분동안끓였으며 n-butanol 을넣은후 2000 rpm에서 15분원심분리하여상층액을 535 nm에서흡광도를측정하였다. 1,1,3,3,-tetraethoxypropane 농도에따른 TBA값으로표준곡선을이용하여각시료의지질과산화물의함량을계산하였다. 간내항산화효소의활성측정간내항산화효소의활성을측정하기위하여간조직을균질화한후, superoxide dismutase (SOD), catalase, glutathione peroxidase (GSH-px) 활성을 kit (Cayman, CA, USA) 을이용하여측정하였다. SOD는간균질액을 1.500 g에서 5분간원심분리한뒤상층액을 radical detector에넣은후 zanthine oxide를넣어 440~460 nm에서흡광도를측정하였다. 표준용액의흡광도로표준곡선을그린뒤각시료의 SOD 활성을계산하였다. Catalase 의활성은간균질액을 10,000 g에서 15분간원심분리한뒤얻어지는상층액을이용하여실험하였다. formaldehyde standard와시료에 assay buffer, methanol, hydrogen peroxide를넣고 20분간실온에서방치후 potassium hydroxide와 purpald를넣고 10 분간배양하고 potassium periodate를넣어 540 nm에서흡광도를측정하였다. Catalase 활성도표준곡선을이용하여 계산하였다. GSH-px는 10,000 g에서 15분간원심분리하여얻어지는간균질액의상층액을 assay buffer와 cosubstrate에 Cumene Hydroperoxide를넣은후 340nm에서흡광도를시간대별로찍어각시간에따른흡광도의변화를계산하여 GSH-px의활성을측정하였다. 통계분석모든실험결과는평균 ± 표준오차로나타내었다. 각실험군에대한유의차검정은 SPSS 21.0 (SPSS Inc., Chicago, IL, USA) 을사용하여일원분산분석 (ANOVA) 을실시한후, p < 0.05 수준에서 Tukey s multiple range test를이용하여유의성을검증하였다. 결 Jaceosidin이체중변화및식이효율에미치는영향 Jaceosidin의섭취가체중및식이효율에영향을미치는지본알아본결과는 Table 2와같다. 섭취전체중은모든군에서유의한차이가없었으나실험후 STZ로당뇨병을유도한군의체중이 Normal과비교하여유의하게감소하였다. 당뇨군에서는 Jaceosidin의섭취유무에따라체중의차이가없는것으로나타났으나, jaceosidin을섭취한군의체중이 D-Control보다다소높은경향을보였다. 8주간체중증가량을측정한결과 Normal은 2.2 g의체중이증가하였으나, 당뇨군의경우체중이감소하는것으로나타났으며, D-0.02% 의경우 Normal군보다체중이유의적으로감소한것으로나타났다. 식이섭취량의경우, Normal의섭취량이당뇨군에비해유의적으로적은것으로나타났으며, 당뇨군중에서는 D-Control의섭취량이 D-0.005% 에비해유의적으로많은것으로나타났다. 식이효율은정상군이당뇨군에비해유의적으로높게나타났으며, 당뇨군간에는유의적인차이를보이지않았다. Jaceosidin이장기무게에미치는영향 Jaceosidin이장기무게에영향을미치는지알아보기위해간, 비장, 신장, 지방조직의무게를측정한결과는 Table 과 Table 2. Effects of jaceosidin on changes in body weight, food intake, and food efficiency ratio in STZ-induced diabetic mice Groups Initial body weight (g) Final body weight (g) Food intake (g/day) Total body weight gain (g/8 weeks) FER 2) Normal D-Control D-0.005% D-0.02% 25.24 ± 1.68 1) 22.70 ± 3.31 1 23.16 ± 1.72 1) 23.98 ± 2.45 1) 27.44 ± 3.13 a 21.90 ± 2.88 b 22.36 ± 1.27 b 22.48 ± 3.53 b 2.97 ± 0.49 c 5.10 ± 0.57 a 4.30 ± 0.84 b 4.67 ± 0.70 ab 2.20 ± 2.25 a -0.80 ± 3.82 ab -0.80 ± 2.63 ab -1.50 ± 2.15 b 1.13 ± 0.52 a -0.16 ± 0.46 b -0.26 ± 0.41 b -0.46 ± 0.23 b 1) Mean ± SEM 2) FER: Food efficiency ratio Values not sharing the same letter were significantly different (p < 0.05).
316 / Jaceosidin 에의한항당뇨효능 Table 3. Effects of jaceosidin on organ weight in STZ-induced diabetic mice Group Normal D-Control D-0.005% D-0.02% Tissue Weight (g) Liver Spleen Kidney Epididymal adipose tissue Perirenal adipose tissue 1.132 ± 0.093 1) 0.090 ± 0.026 a 0.351 ± 0.033 0.709 ± 0.273 a 0.231 ± 0.097 a 1.047 ± 0.231 0.057 ± 0.021 b 0.364 ± 0.058 0.138 ± 0.148 b 0.017 ± 0.042 b 1.095 ± 0.089 0.054 ± 0.008 b 0.347 ± 0.058 0.210 ± 0.122 b 0.034 ± 0.047 b 1.061 ± 0.143 0.057 ± 0.016 b 0.369 ± 0.048 0.174 ± 0.191 b 0.045 ± 0.076 b 1) Mean ± SEM Values not sharing the same letter were significantly different (p < 0.05). Table 4. Effects of jaceosidin on liver function in STZ-induced diabetic mice Normal D-Control D-0.005% D-0.02% GOT (U/L) 65.8 ± 8.9 1) 131.8 ± 66.6 93.6 ± 18.1 132.0 ± 70.7 GPT (U/L) 16.6 ± 3.3 55.2 ± 36.2 32.4 ± 5.4 48.7 ± 40.5 1) Mean ± SEM Values not sharing the same letter were significantly different (p < 0.05). 3과같다. 간과신장의무게는그룹간유의적인차이를보이지않았으나, 비장과부고환지방, 신장주위지방조직의무게는당뇨군이 Normal군에비해유의적으로적었다. Jaceosidin의섭취에따른조직무게의변화는 D-Control군과비교하여유의적인차이를보이지않았으나부고환지방과신장주위지방조직의경우 D-Control에비해다소높은경향을보였다. Jaceosidin이간기능에미치는영향 Jaceosidin의섭취가간기능에미치는영향을알아보기위해혈장 GOT, GPT 활성을측정한결과 (Table 4), GOT와 GPT의활성은그룹간에유의한차이가없었으나 Normal 군이활성이당뇨군에비해낮게나타났으며, jaceosidin 섭취는간기능에영향을주지않는것으로나타났다. Jaceosidin이공복혈당, 내당능및인슐린내성에미치는영향제1형당뇨병에서 jaceosidin의섭취가공복혈당, 내당능및인슐린내성에미치는영향을알아보기위해공복혈당과당부하검사, 인슐린내성을측정하였다 (Fig. 1). 공복혈당의경우당뇨군의혈당이 Normal군에비해유의적으로높았으며, jaceosidin을섭취시킨군이 D-Control에비해감소하는경향을보였다. 특히 D-0.005% 군은 D-Control에비해공복혈당수준이유의적으로낮았다 (Fig. 1A). 7주간의 jaceosidin의섭취가당뇨시내당능에미치는영향을측정한결과는 Fig. 1B와 1C에제시되었다. Normal군의공복혈당이당뇨군보다유의적으로낮았으며, 당뇨군간에는유의적인차이를보이지않았다. 또한, 정상군의경우포도당을경구투여한이후 15분경에최대혈당수치를 보였으며, 이후감소하여 120분이후에는공복혈당과비슷한수치가되었다. 그러나, 당뇨군의경우포도당투여후 15~30분에최고혈당치를보였고, jaceosidin의섭취에따른내당능의차이는보이지않았다. Jaceosidin의섭취에따른인슐린저항성을측정한결과 (Fig. 1D, 1E), Normal 군의경우혈당이서서히감소하여 60 분에가장최저치를보였고, 그이후서서히증가되었다. 당뇨군의경우인슐린처치 15분이후부터혈당이감소하기시작하였으며, 120분까지공복혈당보다낮은수준으로혈당이유지되었다. 또한, 인슐린저항성의경우 jaceosidin 섭취군과 D-Control간에유의적인차이는없었으나, D- 0.002% 군의인슐린내성이낮은경향을나타내었다. Jaceosidin이과산화지질생성및체내항산화체계에미치는영향 Jaceosidin 섭취가당뇨병으로인해야기되는산화적스트레스와항산화체계에미치는영향을측정한결과는 Table 5와같다. Jaceosidin의섭취가당뇨병으로인한산화적스트레스에미치는영향을간내과산화지질함량으로측정한결과모든군에서유의적인차이를보이지않았다. 항산화효소활성의경우, Normal군의활성이 D- Control보다높게나타났다. 또한, 당뇨병으로인해낮아진항산화효소의활성은 jaceosidin의섭취에의해높아지는경향을보였다. GSH-px의경우, D-0.005% 와 D- 0.02% 군의활성이 D-Control과비교하여유의적으로높게나타났으며, Normal 군과는차이를보이지않았다. 또한, catalase의경우 jaceosidin을섭취한군의활성이 D- Control보다높은경향을보였으나, 유의적인차이는없는것으로나타났다.
Journal of Nutrition and Health (J Nutr Health) 2014; 47(5): 313 ~ 320 / 317 Fig. 1. Effects of jaceosidin intake on fasting blood glucose, oral glucose tolerance test and insulin tolerance test in STZ-induced mice. A: Fasting blood glucose levels after 8 weeks of diet, B: Oral glucose tolerance test (OGTT) was performed at 6 week of diet. C: Area of under curve (AUC) was calculated based on OGTT results. D: Insulin tolerance test (ITT) was performed at 7 week of diet. E: AUC was calculated based on ITT results. 고찰 당뇨병증상이나당뇨병합병증은혈당이높게유지되면서과다하게생성된유리기로인해항산화시스템의기능이감소되어항산화체계의균형이깨지고, 이로인해세포 막의지질과산화물과최종당산화물 (AGEs) 이생성되고축적됨으로써야기된다고보고되고있다. 10,12 또한, 산화적스트레스시전사인자인 NF-κB의활성이증가되어염증인자의발현을증가시킴으로써당뇨병증상을더욱악화시키는것으로보고되고있다. 22-26 당뇨로인한고혈당
318 / Jaceosidin 에의한항당뇨효능 Table 5. Effects of jaceosidin on liver lipid peroxidation and antioxidant enzyme activities in STZ-induced diabetic mice Normal D-Control D-0.005% D-0.02% TBARS (µm/mg protein) SOD (U/ml/mg protein) GSH-px (nmol/min/ml) Catalase (nmol/min/ml/mg protein) 0.78 ± 0.05 1) 67.14 ± 8.15 a 694.04 ± 42.53 ab 323.23 ± 13.03 a 0.68 ± 0.04 36.75 ± 12.41 ab 597.82 ± 64.06 b 272.74 ± 16.40 b 0.66 ± 0.67 28.94 ± 12.60 b 761.95 ± 31.01 a 289.54 ± 14.52 ab 0.79 ± 0.62 58.49 ± 8.44 ab 826.57 ± 36.09 a 301.79 ± 11.52 ab 1) Mean ± SEM Values not sharing the same letter were significantly different (p < 0.05). 과과도한산화적스트레스, 염증반응은심혈관계질환, 당뇨성망막증, 당뇨신증, 상처치유억제와같은합병증을유발시키는것으로알려져있다 2. 8,28 따라서, 당뇨병의증상을완화시키기위해혈당을낮출뿐아니라산화적스트레스를완화시킴으로써당뇨병을억제하고그합병증을예방하기위한연구가지속적으로이루어지고있으며, 항산화, 항염증및항암기능 19,29,30 을갖는 jaceosidin이당뇨병의증상을완화시킬수있을것으로사료되는바, 제 1형당뇨병모델을이용하여 jaceosidin이혈당및항산화능에미치는효능을알아보았다. Jaceosidin의항당뇨효능을제1형당뇨모델을이용하여측정한결과, D-0.005% 군의공복혈당이 D-Control에비해유의적으로낮게나타났으며, D-0.02% 군은유의적인차이는없었으나 D-Control에비해감소하는경향을보였다. 여러연구결과에의하면항산화제를섭취하거나항산화효소의발현을증가시키게되면산화적스트레스가감소되어췌장내 β세포의사멸이억제되거나, 간내 glucose transporter의발현이증가되어혈당이감소하고산화적스트레스를저하시킨다고보고되고있다. 31-33 따라서, jaceosidin 에의한혈당감소효과가산화적스트레스를억제함으로써야기되는것인지를알아보기위해간내과산화지질의농도와항산화효소의활성을측정하였다. 그결과, 과산화지질의농도에는유의적인차이가없었으나, GSH-px의활성이 D-Control에비해유의적으로높아졌으며, catalase의활성도높아지는경향을보였다. 따라서, jaceosidin의섭취가과산화지질의생성을억제하지는못하였으나, jaceosidin 자체의항산화력과체내항산화효소인 GSH-px의활성을증가시켜산화적스트레스를억제함으로써공복혈당을감소시킨것으로사료된다. Jaceosidin이함유된약쑥은강력한항산화력이있는것으로보고되고있다. 30 Superoxide anion, hydrogen peroxide, hydroxyl radical과같은유리기의소거능이 positive Control인 butylated hydroxyanisole (BHA) 보다높은것으로나타났으며, 간내지질과산화물의생성을효과적으로감소시키는것으로보고되었다. 34 이러한항산화능은약쑥내다량함유된 flavone인 eupatiolin 과 jaceosidin에의한것으로나타났다. 17 뿐만아니라 jaceosidin은 NF-κB의활성을억제함으로써 pro-inflammatory mediators인 prostaglandin E 2 와 tumor necrosis factor-α, interleukin-1의발현을억제하는것으로보고되고있다. 19 따라서, jaceosidin은외인적, 내인적항산화력을증진시켜산화적스트레스를감소시킬뿐아니라과도한염증반응으로인해악화될수있는당뇨병의증상을효과적으로억제할수있을것으로사료된다. 다른여러연구에서도항산화기능이있는성분이나각종식물추출물의섭취가당뇨병으로인한산화적스트레스를감소시켜공복혈당을감소시키거나, 35,36 혈당과상관없이항산화능을향상시키는것으로보고하고있다. 37 생약제나와송추출물을 1% 섭취시킨군의경우공복혈당을유의적으로감소시키고, 항산화능을향상시켜지질과산화물의생성을억제하는것으로보고되었으며, 36,37 asiatic acid (20 mg/kg body weight) 를제1형당뇨모델에섭취시킨결과, 공복혈당을낮출뿐아니라 GSH와 GSH-px, catalase등체내항산화효소의활성을높이는것으로보고되었다. 36 그러나, 대부분의추출물이나단일성분의항당뇨효능연구에서는총섭취량의 0.5%~1% 또는체중당 20 mg을섭취시켜본연구에서사용한 0.005%~0.02% 보다다량의성분을섭취시켰다. 따라서, 소량의섭취만으로공복혈당을감소시킬뿐아니라항산화효소의활성을증가시키는 jaceosidin이당뇨병을예방하고억제하는데더욱효과적으로활용할수있을것으로사료된다. Jaceosidin 의섭취여부에따른내당능을측정한결과유의적인차이를보이지않았으나인슐린에대한민감성을측정하는 ITT 실험결과에서는 0.02% Jaceosidin을섭취한군의 AUC ( 곡면하면적 ) 가감소하는것으로나타나인슐린민감도가당뇨대조군에비해높은것으로나타났다. 이러한결과는쑥추출물의섭취가인슐린저항성을낮춰당뇨증상을완화시켰다는연구결과 38 와유사한경향을보였다. 제1형당뇨병의경우인슐린을투여하여혈당을낮추기때문에인슐린의민감성을향상시켜주는 jaceosidin과같은기능성성분을섭취한다면적은양의인슐린으로혈당을효율적으로조절할수있을것으로기대된다. STZ에의해유도된당뇨군은정상군과비교하여체중및
Journal of Nutrition and Health (J Nutr Health) 2014; 47(5): 313 ~ 320 / 319 식이효율이낮았다. 이는당뇨병으로인해체내에너지대사에불균형이초래됨으로써발생한것으로, 다량의항산화성분을섭취하게되면체중감소가완화되고, 식이효율이높아진다고보고되고있다. 39,40 그러나본연구에서는항산화능이있는 jaceosidin의섭취가체중감소와식이효율에영향을미치지않는것으로나타났는데, 이는 jaceosidin을식이의 0.005%~0.02% 로섭취시켜다른연구와비교하여적은양 (0.1%~0.5%) 을섭취시켰기때문으로판단된다. 39,40 또한, jaceosidin 섭취에의한간, 비장, 신장, 지방조직의무게를비교한결과당뇨군과비교하여유의적인차이를보이지않았으나당뇨대조군에비해 jaceosidin을섭취한군의지방조직무게가다소높은경향을보였다. 본연구에서는 jaceosidin의섭취가인슐린결핍으로인해발생하는제1형당뇨병에미치는영향을연구함에있어인슐린의민감도를측정하였을뿐혈중인슐린농도나분비능을측정하지못한한계점이있으므로이에대한추가적인연구가필요할것으로생각된다. 결론적으로 jaceosidin 섭취는 STZ로유도한당뇨병모델의공복혈당을감소시키고, 체내항산화효소인 GSHpx의활성을높여주며, 인슐린민감성을증가시키는효과가있어제1형당뇨병을억제하거나당뇨병으로인한합병증을예방할수있을것으로생각되며, 항당뇨기전이나당뇨합병증에대한연구및임상연구등의후속연구가이루어진다면항당뇨기능성성분으로사용될수있을것으로사료된다. 요 본연구는약쑥의기능성성분인 jaceosidin이제1형당뇨병에서혈당과체내항산화상태에미치는영향를조사하고자실시되었다. jaceosidin을식이에농도별 (0.005%, 0.02%) 로혼합시켜준비하였으며, 실험동물은 C57BL/6J 5주령수컷마우스를사용하여정상군 (Normal) 과당뇨군으로나눈뒤당뇨군의마우스는 5일간 streptozotocin (55 mg/kg bw) 을복강으로주사하여당뇨를유발하였다. 당뇨군은당뇨대조군 (D-Control), jaceosidin 0.005% 섭취군 (D-0.005%), jaceosidin 0.02% 섭취군 (D-0.02%) 으로나누어사육하였다. 8주간의식이섭취결과, 당뇨군의체중이 Normal과비교하여유의적으로감소되었고, 식이효율은유의적으로낮아졌으며, jaceosidin의섭취는당뇨로인한체중및식이효율저하에영향을미치지않는것으로나타났다. 제1형당뇨시 jaceosidin의섭취가장기무게와간기능에영향을미치지않는것으로나타났으나, 공복혈당은 D- 0.005% 가 D-Control과비교하여유의하게감소하는것으 약 로나타났으며, 농도의존적은경향은보이지않았다. 당부하검사에서는당뇨군이 normal에비해혈당과 AUC 값이유의적으로높게나타났으나, jaceosidin 섭취에의한차이는없는것으로나타났으며, 인슐린내성검사결과에서는 D-0.02% 가당뇨대조군보다 AUC값이감소하는경향을보였다. Jaceosidin의섭취가당뇨병로인한산화적스트레스에미치는영향을측정한결과, jaceosidin의섭취가과산화지질생성에는영향을미치지않는것으로나타났으나, 항산화효소인 GSH-px의활성은 D-Control과비교하여유의적으로증가시켰다. 이상의결과로미루어볼때, jaceosidin 은제1형당뇨병시간의항산화효소계를활성화시킴으로써공복혈당을낮추고, 인슐린민감성을증가시키는것으로생각되는바당뇨병이나당뇨로인한합병증예방및치료에활용될수있을것으로판단된다. References 1. Lim S, Lee EJ, Koo BK, Cho SI, Park KS, Jang HC, Kim SY, Lee HK. Increasing trends of metabolic syndrome in Korea-based on Korean National Health and Nutrition Examination Surveys. J Korean Diabetes Assoc 2005; 29(5): 432-439. 2. Alberti KG, Zimmet PZ. Definition, diagnosis and classification of diabetes mellitus and its complications. Part 1: diagnosis and classification of diabetes mellitus provisional report of a WHO consultation. Diabet Med 1998; 15(7): 539-553. 3. Pambianco G, Costacou T, Ellis D, Becker DJ, Klein R, Orchard TJ. The 30-year natural history of type 1 diabetes complications: the Pittsburgh Epidemiology of Diabetes Complications Study experience. Diabetes 2006; 55(5): 1463-1469. 4. Baynes JW. Role of oxidative stress in development of complications in diabetes. Diabetes 1991; 40(4): 405-412. 5. Eppens MC, Craig ME, Cusumano J, Hing S, Chan AK, Howard NJ, Silink M, Donaghue KC. Prevalence of diabetes complications in adolescents with type 2 compared with type 1 diabetes. Diabetes Care 2006; 29(6): 1300-1306. 6. Brownlee M. Biochemistry and molecular cell biology of diabetic complications. Nature 2001; 414(6865): 813-820. 7. Loomans CJ, de Koning EJ, Staal FJ, Rookmaaker MB, Verseyden C, de Boer HC, Verhaar MC, Braam B, Rabelink TJ, van Zonneveld AJ. Endothelial progenitor cell dysfunction: a novel concept in the pathogenesis of vascular complications of type 1 diabetes. Diabetes 2004; 53(1): 195-199. 8. Giacco F, Brownlee M. Chapter 35. Pathogenesis of microvascular complications. In: Holt RI, Cockram C, Flyvbjerg A, Goldstein BJ, editors. Textbook of Diabetes, 4th edition. Chichester: Wiley- Blackwell; 2010. p.555. 9. Jacob RA. The integrated antioxidant system. Nutr Res 1995; 15(5): 755-766. 10. Evans JL, Goldfine ID, Maddux BA, Grodsky GM. Oxidative stress and stress-activated signaling pathways: a unifying hypothesis of type 2 diabetes. Endocr Rev 2002; 23(5): 599-622. 11. Yasunari K, Maeda K, Nakamura M, Yoshikawa J. Oxidative
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