Journal of Nutrition and Health (J Nutr Health) 2015; 48(2): 133 ~ 139 http://dx.doi.org/10.4163/jnh.2015.48.2.133 pissn 2288-3886 / eissn 2288-3959 Research Article 갱년기모델유도흰쥐에서갈근과지황복합물및유산소운동이대사이상개선에미치는영향 * 오상아 1 옥향목 1 김혜진 2 이원준 2 권오란 1 이화여자대학교식품영양학과, 1 이화여자대학교체육과학과 2 Effects of a Pueraria lobata-root based combination supplement containing Rehmannia glutinosa and aerobic exercise on improvement of metabolic dysfunctions in ovariectomized rats* Oh, Sang A 1 Ok, Hyang Mok 1 Kim, Hye Jin 2 Lee, Won Jun 2 Kwon, Oran 1 1 Department of Nutritional Science and Food Management, Ewha Womans University, Seoul 120-750, Korea 2 Department of Exercise Science, College of Health Sciences, Ewha Womans University, Seoul 120-750, Korea ABSTRACT Purpose: There is a fair amount of evidence indicating that increased risk of obesity and insulin resistance is associated with postmenopausal state, but can be modulated by diet and exercise. In this study, we explored whether a Pueraria lobata root-based supplement containing Rehmannia glutinosa (PR) and/or aerobic treadmill exercise can modify the metabolic changes associated with estrogen deficiency. Methods: Seventy rats were randomly assigned to the following groups for 8 weeks (n=10 per group): SHAM, sham-operated; PR0, ovariectomized (OVX) control; PR200, OVX with PR200 mg/kg B.W; PR400, OVX with PR400 mg/kg B.W; EPR0, OVX with exercise; EPR200, OVX with exercise and PR200 mg/kg B.W; EPR400, OVX with exercise and PR400 mg/kg B.W. Results: OVX induced significant increases in body weight, food intake, fat mass, LDL-cholesterol, and fasting blood glucose, confirming induction of menopausal symptoms. PR supplementation or exercise significantly suppressed the above mentioned changes through different regulatory elements in adipose tissue: PR supplement upregulated adiponectin gene expression and aerobic exercise upregulated adiponectin and insulin receptor gene expression and a combination of PR supplement and aerobic exercise showed an additive effect on adiponectin gene expression. Conclusion: Taken together, the results of this study suggest that PR supplement has a potential to provide health benefits in OVX rats through leptin and adiponectin secretion. In addition, the data suggest that combination of exercise and PR would have additive effects on metabolic dysfunction associated with estrogen deficiency. KEY WORDS: Pueraria lobata, Rehmannia glutinosa, aerobic exercise, metabolic dysfunction, ovariectomized rats 서 갱년기는여성의삶에있어결정적인시기이다. 신체조직이노화하는과정에서난포자극호르몬은증가되나에스트로겐분비가저하되므로에스트로겐수용체를갖는조직및기관이영향을받아체지방량과인슐린저항성이증가하고간, 지방조직, 근육의항상성유지가깨어진다. 1 론 이로써갱년기여성은당뇨병, 심혈관계질환등의위험이증가되는것으로보고되고있다. 갱년기건강증진을위한방법으로는호르몬보충요법, 기능성식품의섭취, 규칙적인운동을들수있다. 이중호르몬보충요법은부작용의우려가높으므로기능성식품의섭취와규칙적인운동등생활양식의변화가선호되고있다. 2 에스트로겐이심혈관질환과관련이있음은역학적으로 Received: December 3, 2014 / Revised: February 3, 2015 / Accepted: March 27, 2015 *This work was carried out with the support of Cooperative Research Program for Agriculture Science & Technology Development (Project title: Research on New Functional Food Materials with Domestic Agricultural Products, Project No. PJ00845002) Rural Development Administration, Republic of Korea. To whom correspondence should be addressed. tel: +28-2-3277-6860, e-mail: orank@ewha.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 (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.
134 / 갈근 / 지황복합추출물과운동의인슐린저항성 확인되었으며, 3 그기전연구를위해난소절제로에스트로겐분비를감소시킨실험동물을많이사용하여왔다. 조등 4 은난소절제흰쥐는혈관의이완능력이감소되었을뿐아니라형태학적으로전형적인동맥경화증의소견을초래하였다고보고하고있다. 최근에는갱년기의건강유지기능성을검증하는데난소절제갱년기모델유도흰쥐를많이사용하고있다. 5,6 칡 (Pueraria lobata) 은뿌리 ( 갈근 ) 와꽃 ( 갈화 ) 부위가예로부터사용되어왔는데, 특히갈근은고혈압, 관상동맥경화증, 협심증, 노인성당뇨등의치료에사용되어왔다. 갈근에함유된이소플라본류의 90% 이상이 puerarin으로분석되었고, 7 갈근또는갈근에서분리한이소플라본을중심으로항산화효과를보고한연구가가장많다. 8,9 이소플라본은에스트로겐유사활성을나타내어에스트로겐의대체효능을발휘하는것으로알려져있다. 10 또한현삼과식물인지황 (Rehmannia glutinosa) 은뿌리부위가가장활성을지니는부분으로전통적으로몸안에있는기를원활히순환하여혈을보하고, 신진대사기능을회복시키는효과를가지고있다고알려져있다. 11 최근에는지황추출물을중심으로항산화 12 및간보호기능 13 이주로연구되고있다. 지황의기능성분으로는이리도이드배당체화합물인 catalpol과 leonuride 등이알려져있다. 14 이리도이드배당체는지황뿐아니라노니의기능성분으로도알려져있다. 15 그러나갈근, 지황또는갈근과지황복합물의갱년기건강에대한연구는매우드물며, 아울러갈근과지황복합물과운동의병행효과에대한연구는전무한실정이다. 따라서본연구는난소절제를통해인슐린저항성과지질대사이상을유도한모델동물을사용하여갈근과지황복합물섭취와유산소운동이이상지질혈증과인슐린저항성에대한개선효과가있는지를알아보고그기전을밝히고자하였다. 연구방법 시험물질시험물질인갈근 (Pueraria lobata) 과지황 (Rehmannia glutinosa) 복합물은뉴메드 (Seoul, Korea) 에서공급받았다. 갈근및지황각추출물을제조하고혼합하여갈근과지황복합물을제조하였다. 간략히정리하면갈근과지황 100 g을각각세절하여 10배중량의물을첨가하여반복추출하였다. 추출물은여과하여감압농축후동결건조하여분말상태로만들었다. 갈근및지황분말은같은중량비로혼합하여복합물을제조하였다. 지황추출물의수율 은 53.86% 이며, 갈근추출물의수율은 54.19% 이다. 갈근과지황복합물은제조공정상의표준화를위해 HPLC로 puerarin과 catalpol을분석하였다. 갈근의 puerarin 함량평균은 42.1 mg/g이었으며, 지황의 catalpol 함량평균은 9.3 mg/g 으로분석되었다. 운동프로토콜트레드밀 (motor-driven rodent treadmill, Cornella (BCN), Panlab, Spain) 을이용하여오전일정한시간에주 5회운동을실시하였다. 운동강도는점차증가하여 5주부터고강도의운동을지속하기위해처음 5주동안에는경사없이매분 15 m의속도로하루에 15분운동을실시하였으며, 이후 3주동안에는 10% 경사에서매분 26 m의속도로매일 60분운동을실시하였다. 16,17 실험동물및식이본실험에사용된실험동물은 8주령 Sprague-Dawley계암컷흰쥐 (180~200 g) 이며, 중앙실험동물 (Seoul, Korea) 로부터공급받았다. 일주일간물과표준식이를공급하여사육실환경에적응시켰다. 실험실의사육조건은온도 22 ± 2 o C, 습도 55 ± 5%, 12시간간격으로명암조절을자동유지하였다. 적응기간후정상대조군 (SHAM, n = 10) 과양측난소절제군 (OVX, n = 60) 으로분류하고마취제 (Zoletil: Rumpun mixture (3:1) 0.2 mg/kg B.W.) 를복강내주입하여마취한후등부위를절개하여양측난소를절제하였다. 수술부위의감염방지를위해포비돈요오드액을수술부위에도포하며일주일의회복기를가졌다. 일주일의회복기후양측난소절제술군은난괴법으로 10마리씩 6군으로분류하였다. 즉, 시험물질과운동의 2요인디자인으로운동부하없이시험물질의용량 (0, 200, 400 mg/kg B.W.) 에따라 3군 (PR0, PR200, PR400) 으로나누고, 운동을부하하며시험물질의용량에따라 3군 (EPR0, EPR200, EPR400) 으로총 6군이난소절제술군에포함되었다. AIN-76A 식이 18 를기본식이로제공하였으며, 증류수또는시험물질을매일일정한시간에경구투여하여총 8주간실시하였다. 본연구는이화여자대학교동물실험윤리위원회의승인을받아수행하였다 (IACUC No. 2013-01-075). 체중, 식이섭취량, 채혈및장기적출체중은주 2회일정한시각에측정하였으며, 식이섭취에서오는갑작스런체중의변화를막기위하여, 2시간공복상태에서체중측정을하였고, 식이섭취량은매일일정한시간에측정하여계산하였다. 실험동물을 12시간이상절식시킨후, CO 2 가스로호흡마취하였다. 혈액은복대동
Journal of Nutrition and Health (J Nutr Health) 2015; 48(2): 133 ~ 139 / 135 맥에서채혈하여 EDTA가처리된튜브와별도의처리가없는튜브에넣어 4 o C, 3,500 rpm, 10분동안원심분리한후혈장과혈청을 -80 o C에보관하여분석에이용하였다. 실험동물의간, 근육및지방조직은적출하고생리식염수로세척후여분의수분을제거하고무게를측정한뒤액체질소로급속동결시켜분석시까지 -80 o C에보관하였다. 혈장의생화학적지표분석혈장총콜레스테롤 (TC), 중성지질 (TG), HDL-콜레스테롤 (HDL-C) 농도는측정용 kit (Asan Pharmaceutical, Seoul, Korea) 를사용하여측정하였다. 혈장 LDL-콜레스테롤 (LDL-C) 농도는 LDL-C = TC-HDL-C-(TG/5) 의 Freidewald 19 의방법에따라계산하였다. 공복혈당 (FBG), 인슐린, C-peptide 농도는측정용 kit (Asan Pharmaceutical, Seoul, Korea 및 Mercodia, Uppsala, Sweden) 을사용하여측정하였다. 간조직의지질분석간조직의지방함량은 Folch방법 20 을사용하여분석하였다. 클로로포름을사용하여간조직에서지질을녹여낸후, 유기물을증발시키고남은지질의무게로계산한후 TG 와 TC 농도를측정하였다. 유전자발현분석 TRIzol (Invitrogen, San Diego, CA, USA) 을이용하여간과지방조직에서 total RNA를추출하였다. BioSpec nano spectrophotometer (Shimadzu, Kyoto, Japan) 를사용하여 RNA 함량을측정하고, cdna reverse transcription kit (Applied Biosystem, Foster City, CA, USA) 를사용하여 cdna를합성하였다. mrna 측정을위하여 Step-One- Plus RT-PCR System (Applied Biosystem Foster City, CA, USA) 을사용하여합성된 cdna sample을증폭시켰다. 각유전자에대한 primer 로는렙틴 [Lep; Rn00565158_m1], 아디포넥틴 [Adipoq; Rn00595250_m1], insulin receptor [IR; Rn00565158_m1], 및 β-actin [Actb; Mm00667869_ m1] 을사용하였다. mrna 발현수준은 CT 방법으로계산하였으며, 모든데이터는 β-actin 값에대한상대적인양이다. 통계분석모든실험결과는 Statistical Analysis Systems package version 9.3 (SAS Institute, Cary, NY, USA) 을이용하여분석하였으며, 평균 ± 표준오차 (S.E.) 로나타내었다. 정상대조군과난소절제군간의비교를위하여 Student s t-test 를수행하였다. 난소절제군에서물질보충및운동간차이검증을위하여이원변량분산분석 (two-way analysis of variance) 을실시하여두가지요인중주효과와물질보충과운동부하의교호작용에대한실험군간차이를검증하였다. 또한, 유의수준은유의성이있는경우사후검증으로 Duncan's multiple range test를사용하였다. 모든통계적유의성은유의확률이 5% 미만 (p < 0.05) 일때유의한것으로평가하였다. 결 체중, 식이섭취량및지방조직무게난소절제, 시험물질섭취및유산소운동이체중및식이섭취량에영향을미치는지알아본결과는 Table 1과같다. 정상대조군에비해난소절제군에서 8주후최종체중, 식이섭취량이유의적으로증가하였다 (p = 0.0001, p = 0.0217). 체중과식이섭취량의증가는 PR 보충 (p = 0.003, p < 0.0001) 과운동부하 (p = 0.017, p = 0.002) 에따라유의하게억제되었다. 복막후지방의무게도정상대조군에비해난소절제군에서유의적으로증가하였고 (p = 0.043), PR 보충 (p < 0.0001) 과운동부하 (p = 0.012) 에따라유의 과 Table 1. Effects of PR and aerobic exercise on final body weight, food intake, and fat mass in ovariectomized Groups SHAM OVX Two-way ANOVA PR0 PR200 PR400 EPR0 EPR200 EPR400 PR Ex PR x Ex Final body weight (g) 295.9 ± 9.61 352.4 ± 6.3 *a 331.7 ± 6.1 b 286.4 ± 8.5 cd 313.8 ± 9.2 b 293.4 ± 6.2 c 268.5 ± 5.5 d 0.003 0.017 0.260 Food intake (g) 17.52 ± 0.64 19.3 ± 0.3 *a 17.4 ± 0.4 b 15.0 ± 0.5 d 16.8 ± 0.5 bc 15.8 ± 0.4 cd 14.6 ± 0.3 d < 0.0001 0.002 0.423 Retroperitoneal fat (g) 5.32 ± 0.80 7.4 ± 0.5 *a 5.4 ± 0.5 b 2.8 ± 0.3 de 5.0 ± 0.5 bc 3.9 ± 0.5 cd 2.2 ± 0.2 e < 0.0001 0.012 0.723 Perirenal fat (g) 1.64 ± 0.17 1.9 ± 0.3 a 1.8 ± 0.2 a 1.0 ± 0.1 b 1.7 ± 0.1 a 1.1 ± 0.1 b 0.8 ± 0.1 b 0.001 0.081 0.254 Values are expressed as Means ± SE. SHAM, sham operated; OVX, ovariectomized; PR, Pueraria lobata-root based combination supplement containing Rehmannia glutinosa; Ex, exercise; PR0, ovariectomized; PR200, ovariectomized with PR200 mg/kg B.W; PR400, ovariectomized with PR400 mg/kg B.W; EPR0, ovariectomized with exercise; EPR200, ovariectomized with exercise and PR200 mg/kg B.W; EPR400, ovariectomized with exercise and PR400 mg/kg B.W. *p < 0.05 SHAM vs. PR0. Values not sharing the same letter were significantly different in the OVX group at p < 0.05.
136 / 갈근 / 지황복합추출물과운동의인슐린저항성 하게감소하였다. 그러나신장주변지방의무게는정상대조군에비해높아지는경향이있었으나통계적으로유의하지않았으며, 운동부하보다는 PR 보충에영향을받아난소절제군중에서 PR400, EPR200, EPR400군은신장주변지방의무게가유의하게감소되었다 (p = 0.001). 않은것으로관찰되었다. 그러나난소절제군에서총지방과 TG는 PR 보충에영향받아 (p < 0.0001, p = 0.003), PR200, PR400, EPR200, EPR400군에서유의하게감소하였으며, 운동만수행한 EPR0군에서는유의한영향이나타나지않았다. 혈중지질및간조직지질수준공복혈액에서측정한혈중지질및당지표는 Table 2와같다. 정상대조군에비해난소절제군에서혈중 TG 및 TC 수준은의미있게변화되지않았다. 그러나, 난소절제군중에서 TG는 PR 보충 (p = 0.015) 과운동부하 (p = 0.018) 에영향을받아 EPR400군에서 TG 수준이유의하게감소한반면, TC는 PR 보충에영향을받아 PR200, PR400, EPR200, EPR400군에서유의하게감소하였다 (p < 0.0001). 한편 LDL-C과공복혈당은정상대조군에비해난소절제군에서유의적으로증가하였다 (p = 0.0043, p = 0.0085). 난소절제군중에서는 PR 보충에영향을받아 PR200, PR400, EPR200, EPR400군의 LDL-C와공복혈당증가가유의하게억제되었다 (p < 0.0001, p = 0.0001). 그러나운동만부여한 EPR0군은 LDL-C와공복혈당증가에유의한영향을나타내지못하였고, 공복혈당에서는 PR 보충과운동부하의교호작용이있었다 (p = 0.024). 한편인슐린과 C-peptide 수준은난소절제, PR 보충및운동에의해모두변화되지않았다. 간에서총지방, TC, TG를측정한결과, 정상대조군에비해난소절제군에서이들지표에유의한변화가나타나지 지방조직의렙틴, 아디포넥틴유전자발현지방조직에서렙틴, 아디포넥틴의유전자발현을측정하였다 (Fig. 1). 정상대조군에비해난소절제군에서렙틴유전자의발현은유의하게증가하였으나 (p = 0.0454), 아디포넥틴유전자발현은통계적으로유의한변화가관찰되지않았고 PR 보충및운동부하에서도차이를보이지않았다. 반면아디포넥틴유전자발현은 PR 보충 (p = 0.028) 과운동부하 (p = 0.003), 그리고 PR 보충과운동부하의교호작용 (p = 0.013) 에모두영향받아 EPR200, EPR400군에서유의하게증가하였다. 간및지방조직의인슐린수용체유전자발현간및지방조직에서인슐린수용체의유전자발현을측정한결과는 Fig. 2에제시하였다. 간조직에서는난소절제, PR 보충, 유산소운동여부에의해인슐린수용체유전자발현이변화되지않았다. 지방조직에서도정상대조군에비해난소절제군에서인슐린수용체유전자발현이유의하게변화되지않았으나, 난소절제군은운동부하의영향 (p = 0.011) 으로 EPR0, EPR400군에서유의하게증가한것으로관찰되었다. Table 2. Effects of PR and aerobic exercise on lipid and glucose metabolism in ovariectomized rats Groups SHAM OVX Two-way ANOVA PR0 PR200 PR400 EPR0 EPR200 EPR400 PR Ex PR x Ex Fasting blood TG (mg/dl) 128.8 ± 20.6 104.4 ± 8.0 a 89.7 ± 8.8 a 89.7 ± 8.8 a 90.2 ± 6.7 a 82.1 ± 7.1 ab 63.7 ± 3.8 b 0.015 0.018 0.726 TC (mg/dl) 133.0 ± 8.7 158.5 ± 10.5 a 114.4 ± 6.0 cd 92.4 ± 6.6 d 140.9 ± 6.5 ab 117.8 ± 8.3 bc 117.1 ± 5.2 bc < 0.0001 0.175 0.113 HDL-C (mg/dl) 51.5 ± 3.8 48.1 ± 4.8 a 39.4 ± 3.8 a 25.3 ± 3.2 b 48.6 ± 3.8 a 45.8 ± 3.2 a 38.4 ± 3.8 a 0.001 0.169 0.101 LDL-C (mg/dl) 52.8 ± 6.0 85.4 ± 7.8 *a 52.7 ± 4.7 c 43.3 ± 5.0 c 73.3 ± 3.8 ab 55.5 ± 7.0 bc 60.5 ± 3.5 bc < 0.0001 0.072 0.304 Glucose (mg/dl) 102.6 ± 5.7 168.2 ± 16.8 *a 104.7 ± 9.3 b 100.7 ± 6.3 b 180.3 ± 25.1 a 88.0 ± 5.4 b 96.5 ± 9.9 b 0.0001 0.126 0.024 Insulin (ug/l) 2.4 ± 0.4 1.7 ± 0.3 1.2 ± 0.2 1.5 ± 0.2 1.8 ± 0.4 1.9 ± 0.4 1.5 ± 0.3 0.150 0.672 0.380 C-peptide (umol/l) 1.9 ± 0.2 1.6 ± 0.2 1.4 ± 0.2 1.5 ± 0.3 1.5 ± 0.2 1.5 ± 0.2 1.0 ± 0.1 0.072 0.230 0.857 Liver (mg/g) Total lipid 26.4 ± 2.1 30.0 ± 3.6 a 21.3 ± 2.1 b 17.8 ± 1.2 b 28.8 ± 3.4 a 20.7 ± 1.6 b 20.0 ± 1.8 b < 0.0001 0.731 0.924 TG 12.6 ± 1.7 15.7 ± 2.5 a 9.6 ± 1.2 bc 6.5 ± 0.9 c 11.9 ± 1.6 ab 9.6 ± 1.6 bc 10.8 ± 1.8 bc 0.003 0.902 0.101 TC 2.1 ± 0.5 3.3 ± 0.8 1.9 ± 0.4 1.5 ± 0.2 1.6 ± 0.4 2.3 ± 0.7 2.1 ± 0.4 0.635 0.459 0.063 Values are expressed as Means ± SE. SHAM, sham operated; OVX, ovariectomized; PR, Pueraria lobata-root based combination supplement containing Rehmannia glutinosa; Ex, exercise; PR0, ovariectomized; PR200, ovariectomized with PR200 mg/kg B.W; PR400, ovariectomized with PR400 mg/kg B.W; EPR0, ovariectomized with exercise; EPR200, ovariectomized with exercise and PR200 mg/kg B.W; EPR400, ovariectomized with exercise and PR400 mg/kg B.W. *p < 0.05 SHAM vs. PR0. Values not sharing the same letter were significantly different in the OVX group at p < 0.05.
Journal of Nutrition and Health (J Nutr Health) 2015; 48(2): 133 ~ 139 / 137 Fig. 1. Effects of PR and aerobic exercise on leptin and adiponectin gene expression the retroperitoneal fat of ovariectomized rats: (A) leptin and (B) adiponectin. SHAM, sham operated; PR0, ovariectomized control; PR200, ovariectomized with PR200 mg/kg B.W; PR400, ovariectomized with PR400 mg/kg B.W; EPR0, ovariectomized with exercise; EPR200, ovariectomized with exercise and PR200 mg/kg B.W; EPR400, ovariectomized with exercise and PR400 mg/kg B.W. * p < 0.05 SHAM vs. PR0. Values not sharing the same letter were significantly different in the OVX group at p < 0.05. Fig. 2. Effects of PR and aerobic exercise on IR gene expression in the organ of ovariectomized rats: (A) liver and (B) adipose tissue. SHAM, sham operated; PR0, ovariectomized control; PR200, ovariectomized with PR200 mg/kg B.W; PR400, ovariectomized with PR400 mg/kg B.W; EPR0, ovariectomized with exercise; EPR200, ovariectomized with exercise and PR200 mg/kg B.W; EPR400, ovariectomized with exercise and PR400 mg/kg B.W; IR, insulin receptor. * p < 0.05 SHAM vs. PR0. Values not sharing the same letter were significantly different in the OVX group at p < 0.05. 고찰 본연구는난소를절제하여인위적으로갱년기를유도한흰쥐에서갈근과지황복합물과운동부여가갱년기대사이상및인슐린저항성에미치는영향을검토하였다. 난소를절제하고 8주간정상식이로사육한흰쥐는정상대조군에비해체중, 식이섭취량, 체지방, 혈중 LDL-C 및공복혈당이유의적으로증가하여갱년기대사이상모델이유도되었음을확인하였다. 비만은갱년기의가장특징적인증상으로, 이는부족해진에스트로겐의분비를보상하기위해말초지방조직에서지방의축적이일으키기때문이다. 21 또한본연구결과, 난소절제후혈중인슐린수준은 변화되지않았으나지방조직에서인슐린수용체유전자발현이감소되었을뿐아니라난소절제후지방조직이비대해지고, 또한지방조직에서렙틴분비가증가된것으로미루어에스트로겐분비저하로인한비만은렙틴및인슐린저항과관련있음을확인하였다. 이는설치류에서난소절제후비만이발생하기전에는당대사이상에큰변화가없지만, 22 난소절제후식욕증진및지방축적으로비만이발생하면대사적장애와인슐린저항성이증가한다는연구결과와일치한다. 23 또한비만에서렙틴이증가하는것은인슐린증가와함께인슐린저항성이증가한다는개념으로받아들여지고있다. 24 한편갈근과지황복합물의보충및운동은서로다른기
138 / 갈근 / 지황복합추출물과운동의인슐린저항성 전으로난소절제로에스트로겐분비가감소되어나타나는대사이상을억제하는데유익한것으로나타났다. 갈근과지황복합물은식이섭취량, 체중, 체지방량, 혈중 LDL- C 및공복혈당증가를유의하게억제하였다. 지방조직에서는아디포넥틴유전자발현을유의하게증가시키는것으로나타났으며, 렙틴유전자발현은요인분석결과유의한영향을미치는요소는아니지만운동부여없이 PR을보충한두군에서렙틴유전자발현이유의하게감소된것으로미루어갈근과지황복합물의작용기전으로지방조직호르몬의변화가중요한것으로사료된다. 본연구에사용한갈근에는이소플라본의일종인 puerarin이함유되어있는데, 이는에스트로겐과유사한활성을내는물질이므로갈근과지황복합물의효과는 puerarin에기인하는것으로추정할수있다. 그러나본연구는 puerarin 화합물을직접시험물질로사용하여결과를얻은것이아니므로단정적으로 puerarin을원인물질로결론지을수없으며, 향후이에대한연구가필요하다. 한편, 운동부하는식이섭취량, 체중및체지방량증가에대해서는갈근과지황복합물과동일하게통계적으로유의한억제효과를나타냈으나, 혈중 LDL-C이나공복혈당증가를억제하는효과는나타내지못하였다. 지방조직에서는운동부하로인슐린수용체와아디포넥틴유전자발현이유의하게증가되었으나, 렙틴유전자발현은변화되지않았다. 이는운동과갱년기증상에대한연구에서운동은인슐린저항을개선하는것으로보고된결과와일치한다. 25 그러나신호전달분자수준의연구는부족한실정이며, 본연구결과역시일부정보만을제공하고있으므로향후이에대한지속적인연구가필요하다. 마지막으로갈근과지황복합물과운동의병행은본연구에서측정된지표들중공복혈당과아디포넥틴유전자발현에서상승효과를나타내는것으로관찰되었다. 이결과는아디포넥틴은인슐린과연계된지방대사를중재하는중요한조절인자라는연구내용과도연결지을수있다. 26,27 이상의결과를종합할때, 갈근과지황복합물보충과유산소운동은난소절제로갱년기를유도한모델동물에서에스트로겐분비저하로야기된체중, 체지방, 혈장지질및혈당대사이상을정상화하는것으로관찰되었다. 이는갈근과지황복합물로인한아디포넥틴유전자발현증가와렙틴저항개선, 유산소운동으로인한아디포넥틴유전자발현및인슐린수용체발현증가, 그리고갈근과지황복합물과유산소운동의병행으로인한아디포넥틴유전자발현증가에대한상승효과에기인된것으로사료된다. 이는실생활적용도가높은흥미로운결과이다. 다만, 본연구에서는난소절제흰쥐에게정상지질수준을제공하 는 AIN-76A 식이를제공하고매일 60분간고강도의유산소운동을부여하였는데, 이는대부분의폐경기여성들이달성하기에어려움이많은과도한운동으로사료된다. 따라서운동강도를달리하였을때비만및대사이상변화차이를규명하는후속연구가진행되고있다. 요 본연구는갈근과지황복합물과유산소운동이난소절제로갱년기를유도한흰쥐에서나타나는지질및당대사이상에미치는영향을확인하였다. 난소절제후 8주간 AIN-76A 식이를급여한결과체중증가량, 식이섭취량, 체지방, 혈장 LDL-C, 공복혈당증가를통해갱년기동물모델이유도되었음을확인할수있었다. 갈근과지황복합물과유산소운동은서로다른기전으로에스트로겐분비저하로나타나는비만및대사이상을정상화하는유익한결과를나타냈다. 지방조직에서확인된관련기전으로는갈근과지황복합물로인한아디포넥틴유전자발현증가및렙틴저항억제, 유산소운동으로인한아디포넥틴및인슐린수용체유전자발현증가, 갈근과지황복합물및유산소운동의병행으로인한아디포넥틴유전자발현증가상승효과가관찰되었다. 이상의결과로갈근과지황복합물과유산소운동의병행은폐경기에스트로겐결핍으로인한대사이상을개선하는효과가있을것으로기대한다. 향후에는폐경기유도동물모델의개선과함께단백질및신호전달분자수준에서추가적인기전연구가필요할것으로사료된다. 약 References 1. Couse JF, Korach KS. Estrogen receptor null mice: what have we learned and where will they lead us? Endocr Rev 1999; 20(3): 358-417. 2. Yeum KH, Lee HJ, Yeo SM, Yoo JH, Kim RH. Relation of regular exercise habit to bone mineral density in Korean climacteric women. J Korean Geriatr Soc 2006; 10(1): 23-30. 3. Manolio TA, Furberg CD, Shemanski L, Psaty BM, O'Leary DH, Tracy RP, Bush TL. Associations of postmenopausal estrogen use with cardiovascular disease and its risk factors in older women. The CHS Collaborative Research Group. Circulation 1993; 88(5 Pt 1): 2163-2171. 4. Cho SH, Cho SH, Moon H, Lee JA, Suh JK, Chung HS. The changes of vascular tone and morphology of rat thoracic aorta induced by oophorectomy. Korean J Obstet Gynecol 1994; 37(7): 1381-1388. 5. Park YS, Kang MS, Kim BK, Kim M. The effect of Eisenia bicyclis extracts on bone tissues in ovariectomized rat. J Korean Soc Food Sci Nutr 2013; 42(1): 33-39.
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