스트레스硏究 : 第 16 卷第 4 號 2008 원저 연자육의항우울효과에대한뇌부위별신경전달물질의변화 * 가톨릭대학교의과대학면역생물학연구소, 가톨릭대학교의과대학통합의학교실, 경희대학교한의과대학생의학교실 김지현 * ㆍ이규섭 ㆍ정혜영 ㆍ심현수 ㆍ배현수 ㆍ김경수 ㆍ심인섭 The Neurochemical Study on Antidepressant Effect of Nelumbinis Semen Extract Ji Hyun Kim*, Kyu-Sop Lee, Hye-Young Joung, Hyun Soo Shim, Hyunsu Bae, Kyung Soo Kim, Insop Shim *Research Institute of Immunobiology, Department of Biomedical Sciences, The Catholic University of Korea, Department of Integrative Medicine, College of Medicine, The Catholic University of Korea, Department of Physiology, College of Oriental Medicine, Kyung Hee University, Seoul, Korea In the previous study, we examined that a variety of extracts of Nelumbinis semen (NS) have an antidepressant effect on the forced swimming test (FST) in the rat, which is widely used as a behavioral screening test for antidepressant activity of new compounds. Depression-like symptom in n-buoh extract showed the strongest inhibitory effect on the immobility time and further fractionated n-buoh with H 2O and MeOH (3:7) fraction (NS37) was the most effective on the immobility time on FST. The purpose of the present study was to observe the neurochemical changes on the anti-depressant actions of unknown compounds in the NS37 extract by using HPLC coupled with electrochemical detection (ECD). The results showed that the NS37-10 (10 mg/kg) has a significant increase of dopamine in cortex, hypothalamus, and hippocampus and a significant decrease of dopamine turnover in hypothalamus and hippocampus. These neurochemical actions of NS37 extract could be responsible for the beneficial effect on depression. (Korean J Str Res 2008;16:393 400) Key Words: Nelumbinis semen, Antidepressant effect, HPLC, Neurotransmitter, Brain 서 최근우울증에대한연구의결과들에의하면우울증에원인이복합적요소를가지고있음은분명하지만, 도파민 (dopamine: DA), 세로토닌 (serotonin: 5-HT), 노르에피네프린 책임저자 : 심인섭, 서울시서초구반포동 505 번지 137-701, 가톨릭대학교의과대학통합의학교실 Tel: 02-590-2971, E-mail: ishim@catholic.ac.kr 접수 : 2008 년 11 월 12 일, 게재승인 : 2008 년 12 월 10 일이논문은 2005 년도정부재원 ( 교육인적자원부학술연구조성사업비 ) 으로한국학술진흥재단의지원을받아연구되었음 (KRF-2005-050-E00005). 론 (norepinephrine: NE) 등을포함하는 monoamine 계열의신경전달물질의기능적불균형혹은결핍에그원인을부여하는 monoamine 가설 은우울증발현의분자기전측면에서오랜기간동안연구되어왔다 (Bunney et al., 1965; Coppen, 1967). 특히 monoamine transporter의기능을막아단기적으로 monoamine의기능을향상시키는효과를가진초기의삼환계항우울제 (tricyclic antidepressant: TCA) 들과 monoamine 산화억제제 (monoamine oxidase inhibitor: MAOI) 가항우울작용이있음은 monoamine 가설을뒷받침하는중요한기반이되고있다 (Fuller, 1995). 1980년대중반부터 fluoxetine, paroxetine, sertralin 등의 selective serotonin reuptake inhibitor (SSRI) 들의등장은 monoamine system이우울증의발현에밀 393
스트레스硏究 : 제 16 권제 4 호 2008 접한관련이있을거라는기존가설을더강화하였다. 이들항우울제가많은환자들에게서긍정적효과를보이는것이사실임에도불구하고적용가능환자군이전체의 50% 이하로한정되어있고, 다양한부작용, 장기투여요구등개선의여지가많을뿐만아니라새로운유형의치료제개발이요구되고있는것또한현실이다. 최근독일에서개발되어국내에도시판되고있는 St. John s Wort (Hypericum perforatum) 은영국에서민간치료제로쓰던것을독일에서개발하였는데이는전문적인항우울제들보다부작용이나안정성에서우위를가지며, 독일의대표적 SSRI계약물인 fluoxetin의 6배에달하는처방율을보이고있다. 이약물은치매치료제인 ginkgo, galantamine 등과더불어생약이임상에서성공한대표적인약물로평가되고있다. 따라서한방약재중에서항우울효과를검색하고증명하는연구가매우의미있으며필요한실정이다. 연꽃의씨앗인연자육은한방에서오랫동안여성의가벼운우울증상이나불안증등의치료를위하여처방되어오던약재로그안정성이입증되어있다 (Jang CG et al., 2004). 그러나기존에알려진연자육의효과는연자육전체 의추출물을대상으로진행되었으며, 연자육의어떤성분이항우울효과를나타내는지에대한연구는수행된바없다. 본연구는 강제수영실험을통한연자육의항우울효과 (Lee KS et al., 2008) 의후속연구로서연자육의투여에의한항우울효과의기전을규명하고자뇌의우울증신경회로로알려진 cortex-hypothalamus-hippocampus (Nestler et al., 2002) 를채택하여부위별 5-HT과 DA의함량을조사하고이들대사산물의측정및대사속도를검토하였다. 재료및방법 1. 실험동물실험에사용된동물은체중 220 250 g의 Sprague Dawley 계수컷흰쥐 ( 오리엔트, 경기도, 한국 ) 로일주일간사육실환경에적응시킨후사용하였다. 사육실온도는 22±2 C, 습도는 55 60% 로유지하고 light-dark cycle이 12시간단위로조절되게한후, 흰쥐용고형사료와물을제한없이공급하였다. Fig. 1. Location of tissue extractions for monoamine analysis. Cortex (COR), hypothalamus (HYP), and hippocampus (HIP). Images scanned and imported directly from the stereotaxic atlas of Paxinos and Watson. 394
김지현외 6 인 : 연자육의항우울효과에대한뇌부위별신경전달물질의변화 2. 연자육의추출및처치실험에사용된연자육은신우약업사 ( 서울, 한국 ) 에서구입하였다. 연자육 4.4 kg을분쇄한후 80% EtOH 20 liter를가한다음 30분간초음파추출 (2510EDTH, Branson, USA) 및여과 (No. 2. Whatman, England) 하고, 이과정을 2회반복하여여과한후 rotary evaporator로 40 C의수욕상에서감압농축 (N-N series, Eyela, 일본 ) 하였다. 80% EtOH 농축액 (192 g, 수득율 4.4%) 은증류수 1 liter를가해녹이고, 용매극성에따라 n-hexane, EtOAc (ethyl acetate), n-buoh, H 2O층으로순차적으로각용매별로 2회씩분획하였다. 각각추출용매별수득율은 n-hexane 분획물 13.85% (26.95 g), EtOAc 분획물 50.22% (10.02 g), H 2O 분획물 29.57% (56.77%) 등이었다. 이들분회물로강제수영검사를실시한결과수영자세의증가가가장큰 n-buoh 분획물을추가분획하였는데, MeOH와 H 2O의비율에따라총 5개로나누어감압농축하였다. 이들분획물의항우울효과의기전을비교검증하기위해강제수영검사를시행하기 24시간, 5시간, 1시간전에각각 1회씩총 3회, 10 mg/kg과 30 mg/kg으로분류하여복강투여하였다. 강제수영검사에서 H 20:MeOH=3:7 조건의추출된물질이부동자세의유의한감소 (10 mg/kg & 30 mg/kg) 및등반자세의유의한증가 (30 mg/kg) 를나타낸실험결과 (Lee KS et al., 2008) 를토대로, 뇌조직에서의 monoamine 정량을위한실험군분류는대조군 (saline 투여 ), NS37-10 군 (H 20:MeOH=3:7에서의추출물, 10 mg/kg 투여 ), NS37-30군 (H 20:MeOH=3:7에서의추출물, 30 mg/kg 투여 ) 으로하였다. 하여여과한후분석에사용하였다. 모든시료는분석할때까지 80 C로즉시냉동보관되었다. 2) 표준약물 : Dopamine (DA), 3,4-dihydroxyphenyl acetic acid (DOPAC), homovanillic acid (HVA), serotonin (5-HT), 5-hydroxyindoleacetic acid (5-HIAA) 를 100 nm 농도로표준용액조제후 inject vol. 5, 10, 20 μl 의 HPLC를실시하여얻은 chromatogram으로부터피크면적을구한후, 이들면적과표준용액의농도를변수로한검량선 (Empower 2 software, Waters, MA, USA) 을이용하여 calibration하였다. 3) 분석기기및분석방법 : 뇌조직중의 monoamine 및그대사산물을정량하기위한 mobile phase의조성은 150 mm sodium phosphate monobasic monohydrate, 1.85 mm octanesulfonic acid, 0.25 mm EDTA, 0.01% triethylamine, 6% acetonitrile, 4% methanol (ph 3.2) 로하였으며 mobile phase는 0.22 μm GV filter (Durapore membrane filter, Millipore, MA, USA) 로여과하여사용하였다. HPLC 장비는 Waters 1525 binary HPLC pump로 0.8 ml/min의유속으로흘렸으며, column (nova-pak C18, 3.9 150 mm, 4 μm particle size) 과 coulometric detector (ESA; Coulochem II Model 5200A with guard cell model 5020; 300 mv, microdialysis cell model 5014B; E2+320 mv, E1-100 mv R; 20 na) 를사용하여분석하였다. 3. HPLC에의한중추신경계 monoamine 및대사산물측정 1) 표본준비 : 실험동물을강제수영검사 (Lee KS et al., 2008) 실시후, 단두하여뇌를적출하고, cortex, hippocampus, hypothalamus를적출하여 (Fig. 1) 각각의무게를재고준비된 buffer 150 mm sodium phosphate monobasic monohydrate, 1.85 mm octanesulfonic acid, 0.25 mm EDTA (ethylene diaminetetra acetic acid) 0.01% triethylamine, 0.1% perchloric acid, 10% acetonitrile에담궜다. 각시료를 Bio-vortexer (Bartlesvill, OK, USA) 로 30초간균질화하였다. 이를 bradford 방법으로단백질정량후 micro 17R centrifuge (Micro 17R, Hanil Co. Korea) 로원심분리 (400 g, 15 min) 한후다시원심분리 (27,000 g, 30 min) 하였다. 상층액을채취하여 PVDF filter (target syringe filter, National Scientific, USA:pore size, 0.45 μm ) 를사용 Fig. 2. High performance liquid chromatographic analysis of monoamine neurotransmitters and their metabolites. Monoamine neurotransmitters and their metabolites were separated by HPLC- ECD system. (1) DOPAC: 3,4-dihydroxyphenylacetic acid, (2) DA: dopamine, (3) 5- HIAA: 5-hydroxyindoleacetic acid, (4) HVA: homovanillic acid, (5) 5-HT: 5-hydroxytryptamine (serotonin). 395
스트레스硏究 : 제 16 권제 4 호 2008 Table 1. Monoamine and metabolite levels in cortex (COR), hypothalamus (HYP), and hippocampus (HIP). Brain area group DA (ng/mg prot) DOPAC (ng/mg prot) HVA (ng/mg prot) 5-HT (ng/mg prot) 5-HIAA (ng/mg prot) COR Control NS37-10 NS37-30 HYP Control NS37-10 NS37-30 HIP Control NS37-10 NS37-30 0.705±0.181 1.655±0.240 a 1.082±0.176 2.529±0.727 6.379±0.926 b 1.758±0.296 0.225±0.075 1.043±0.224 a 0.510±0.307 3.301±1.441 4.156±2.336 1.859±0.425 8.146±2.307 10.825±1.719 2.927±0.727 a 1.567±0.092 2.385±0.669 1.750±0.650 1.234±0.365 2.489±0.532 1.756±0.173 1.290±0.245 2.207±0.483 0.642±0.179 0.453±0.047 0.234±0.086 0.180±0.087 a 1.032±0.376 1.616±0.244 1.680±0.258 3.717±2.085 3.281±0.522 2.222±0.841 1.146±0.165 0.875±0.221 2.223±1.304 2.768±0.371 4.020±0.702 3.226±0.230 11.607±3.510 22.633±4.829 6.714±1.198 4.520±1.399 6.743±0.100 5.598±1.040 Data represent mean±s.e.m. a p<0.05, b p<0.01 versus control group. ANOVA followed by the post hoc LSD test. DA: Dopamine, DOPAC: 3,4-dihydroxyphenylacetic acid, HVA: Homovanillic acid, 5-HT: 5-hyroxytryptamine (serotonin), 5-HIAA: 5-hydroxyindoleacetic acid. Fig. 3. Effects of NS37 treatment on the DOPAC/DA, HVA/DA and 5-HIAA/5-HT ratios in the rat cortex. Data represent mean±s.e.m. ANOVA followed by the post hoc LSD test. Not significant. DOPA: 3,4-dihydroxyphenylacetic acid, DA: dopamine, HVA: homovanillic acid, 5-HIAA: 5-hydroxyindoleacetic acid, 5-HT: 5-hyroxytryptamine (serotonin). Fig. 4. Effects of NS37 treatment on the DOPAC/DA, HVA/DA and 5-HIAA/5-HT ratios in the rat hypothalamus. Data represent mean± S.E.M. ANOVA followed by the post hoc LSD test. *p<0.05 versus control group. DOPAC: 3,4-dihydroxyphenylacetic acid, DA: dopamine, HVA: homovanillic acid, 5-HIAA: 5-hydroxyindoleacetic acid, 5-HT: 5-hyroxytryptamine (serotonin). 4. 통계처리실험결과의통계처리는 SPSS ver. 13.0의 one-way ANOVA (for Windows OS) 를이용하였으며, LSD (least significant difference) test를이용해대조군과의유의성을사후검증하였다 (p<0.05). 결과 1. 표준액의 Chromatogram 뇌조직에서의 catecholamines양을표준화하기위해 DOPAC, DA, 5-HIAA, HVA, 5-HT 표준액 (100 nm) 의 chromatogram을그린결과각각의 retention time은 3.8 min (DOPAC), 4.9 min (DA), 5.7 min (5-HIAA), 8.2 min (HVA), 11.6 min (5-HT) 였다 (Fig. 2). 2. Cerebral cortex내 DA와 5-HT 함량및대사변화 Cerebral cortex에서의 catecholamines양을측정한결과는다음과같다 (Table 1). DA의함량은전반적으로대조군 (0.705±0.181 ng/mg prot) 에비해 NS37 투여시증가되었으며 [F(2,15)=4.820, p=0.027], NS37-10군 (1.655±0.240 ng/mg 396
김지현외 6 인 : 연자육의항우울효과에대한뇌부위별신경전달물질의변화 우대조군 (3.717±2.085 ng/mg prot) 에비해 NS37 투여에의한변화를관찰할수없었으며 [F(2,15)=0.491, p=0.623], 5- HIAA/5-HT 교체율 (Fig. 4) 또한유의미한변화가없었다 [F(2,15)=0.046, p=0.955]. 4. Hippocampus 내 catecholamine 함량및대사변화 Fig. 5. Effects of NS37 treatment on the DOPAC/DA, HVA/DA and 5-HIAA/5-HT ratios in the rat hippocampus. Data represent mean± S.E.M. ANOVA followed by the post hoc LSD test. *p<0.05, p<0.01 versus control group. DOPAC: 3,4-dihydroxyphenylacetic acid, DA: dopamine, HVA: homovanillic acid, 5-HIAA: 5-hydroxyindoleacetic acid, 5-HT: 5-hyroxytryptamine (serotonin). prot) 은대조군에비해유의한증가를보였다 (p<0.05). 한편 A 대사속도를나타내는 DOPAC/DA 교체율과 HVA/DA 교체율 (Fig. 3) 은 NS37 투여시감소하는경향을나타내었으나, 통계적으로유의한변동을나타내지못하였다. 5-HT함량의경우전반적으로대조군 (1.032±0.367 ng/mg prot) 에비해 NS37 투여시함량은증가하고 [F(2,15)=1.364, p= 0.290], 5-HIAA/5-HT 교체율 (Fig. 3) 은감소하는경향을나타내었으나 [F(2,15)=1.581, p=0.243] 통계적으로유의한변동을보이지않았다. 3. Hypothalamus 내 DA 와 5-HT 함량및대사변화 Hypothalamus에서의 catecholamines양을측정한결과는다음과같다 (Table 1). DA의함량은대조군 (2.529±0.727 ng/mg prot) 에비해 NS37 투여시변화가관찰되었는데 [F(2,15)= 13.326, p=0.001], NS37-10군 (6.379±0.926 ng/mg prot) 은대조군에비해유의한증가를보였다 (p<0.01). DOPAC의함량도대조군 (8.146±2.307 ng/mg prot) 에비해 NS37 투여시변화가관찰되었는데 [F(2,15)=7.414, p=0.007], NS37-30군 (2.927±0.727 ng/mg prot) 은대조군에비해유의한감소를보였다 (p<0.05). 한편 DA 대사속도를나타내는 DOPAC/DA 교체율 (Fig. 4) 의경우 NS37 투여시감소하였는데 [F(2,15)= 4.337, p=0.036], NS37-10, NS37-30군모두통계적으로유의미한감소를보였다 (p<0.05). 하지만 HVA/DA 교체율 (Fig. 4) 은 NS37 투여시감소하는경향을보였으나유의한변동을나타내지못했다 [F(2,15)=1.613, p=0.237]. 5-HT함량의경 Hippocampus에서의 catecholamines양을측정한결과는다음과같다 (Table 1). DA의함량은대조군 (0.225±0.075 ng/mg prot) 에비해 NS37 투여시변화가관찰되었는데 [F(2,15)= 2.611, p=0.111], NS37-10군 (1.043±0.224 ng/mg prot) 이대조군에비해유의한증가를보였다 (p<0.05). HVA 함량도대조군 (0.453±0.047 ng/mg prot) 에비해 NS37 투여시변화가관찰되었는데 [F(2,15)=2.624, p=0.110], NS37-30군 (0.180± 0.087 ng/mg prot) 은대조군에비해유의한감소를보였다 (p<0.05). 한편 DA 대사속도를나타내는 DOPAC/ DA 교체율 (Fig. 5) 의경우 NS37 투여시감소하였는데 [F(2,15)= 6.379, p=0.012], NS37-10군에서통계적으로유의미한감소를보였다 (p<0.01). HVA/DA 교체율 (Fig. 5) 또한 NS37 투여시감소하였는데 [F(2,15)=2.729, p=0.102], NS37-10군에서유의한변동을나타냈다 (p<0.05). 5-HT함량의경우는대조군 (1.146±0.165 ng/mg prot) 에비해 NS37 투여에의한변화를관찰할수없었으며 [F(2,15)=0.736, p=0.498], 5- HIAA/5-HT 교체율 (Fig. 5) 또한유의미한변화가없었다 [F(2,15)=1.883, p=0.191]. 고찰선행연구 (Lee KS et al., 2008) 에서보고되었던강제수영검사에서연자육의용매별추출물들의부동시간의감소효과를확인하고그중부동자세의감소및수영자세에서효과를나타낸 n-buoh 분획을선택하여추가적으로 H 2O과 MeOH의비율에따라 5개의물질을분리하여강제수영검사를재실시한후 H 2O:MeOH=3:7에서분리된물질 (NS37) 에서부동자세감소및등반자세증가를보인결과를바탕으로, 흰쥐에게 NS37을투여시뇌조직에서의 monoamine neurotransmitter 함량및대사의변화를구명하기위해 DA, 5-HT 및대사산물을 1회에분석할수있는 HPLC 방법을이용하였다. NS37 (10 mg/kg) 의투여시 cortex, hypothalamus, hippocampus 내의 DA 함량은대조군에비해의미있게증가하였으며, 397
스트레스硏究 : 제 16 권제 4 호 2008 실제 DA 대사속도를표시해줄수있는 DOPAC/DA 교체율은 hypothalamus와 hippocampus에서, HVA/DA 교체율은 hippocampus에서유의하게감소하였다. 5-HT 함량및대사속도는통계적인유의미한변화가관찰되지않았다. 이러한결과는연자육의항우울효과가 5-HT의작용으로나타난다는보고 (Kang MK et al., 2005) 와달리, DAergic neuron의활성화를통해부동시간의감소효과를나타낸다고볼수있다. DA계에대한 5-HT와 DA의상호작용의기전은실험을통해증명되었으나 (Borsini et al., 1985), Kang MK et al. 의실험에서는 DA 함량을측정하지않았으며우울증동물모델의차이가있으므로추후반드시검증이필요하리라생각된다. DA와우울증과의관련을설명하는여러연구들이있는데본연구와유사한동물모델을이용한우울증에서 DA와의연관을설명하려는시도들로 DA agonist 투여를통해부동시간감소효과를관찰하였고, haloperidol에의해이런효과가역전됨을관찰하였다 (Duterte et al., 1988). 우울증의다른동물모델인 sucrose consumption model을통해삼환계항우울제대신 D1 수용체 antagonist인 SCH-23390 혹은 D2 수용체 antagonist인 sulpiride를주면항우울제에의해개선된행동이역전됨을관찰하였으며삼환계항우울제의작용이 DA계의활성화에있음을제시했다 (Sampson et al., 1990). DA과우울증과의관계를설명하는임상적증거로서몇가지우울증유형에있어강력한 DA계의개입의증거가있고, Parkinson병과우울증과의증상에있어서의유사성, 계절성정동장애의 DA계의이상등을들수있다 (Brown et al., 1993). 생화학적증거로서자살을시도했던우울증환자를대상으로저하된 DA 회전율을시사하는뇌척수액에서의 HVA 연구 (Gjerris et al., 1987; Roy et al., 1992), DA 대사물질연구, 약리학적인증거로서우울증의치료에있어서 DA계에영향을끼치는항우울제의효과등이연구되어져있고 (Barber et al., 1985; Wolfe et al., 1990), 정신병적우울증에서 dopamine-ß-hydroxylase의활성도가감소된것을관찰하였다 (Rihmer et al., 1990). 5-HT계도연관되어있다는연구들이있는데 5-HT1B 수용체 agonist가 desipramine으로인한부동시간전소효과를역전시킨다고하였고 (Cervo et al., 1989), 선택적 5-HT1A 수용체 agonist의 DA transmission 의활성화를통해부동시간감소효과를관찰했었고 (Chojnacka et al., 1991), 5-HT1C 수용체 antagonist가 imipramine에의한부동시간감소효과를차단한다는것을보고하였다 (Borsini et al., 1991). 최근에서천연물을이용한항우울제개발을위해많은 연구가이루어지고있다. 이중약 2000년전부터상처치유등에널리사용되어온 Hypericum perforatum이라는식물에서추출한 St. John s Wort 약품이기존항우울약물을대체할수있는효과를가지고있으면서도부작용이적음이밝혀지면서환자를대상으로한대규모 Cohort 연구가이루어지고있고 (Linde et al., 2008), 한랭유영스트레스후소요산을투여받은백서는대조군에비해 prefrontal cortex와 hypothalamus에서 DA의증가가관찰되었으며 hippocampus 에서는 5-HT의유의한증가가있다는보고가있었다 (Lee JH et al., 1998). 본연구에서살펴본연자육내구성물질의항우울효과를보면이들뇌부위에서비록 5-HT의변동은관찰되지않았으나, DA의함량증가및대사속도감소를나타냈기에연자육이항우울작용을갖는물질을포함하고있을가능성을시사한다. 최근급격히발달한뇌 imaging을통한해부학적연구는제한적이나마많은뇌영역들이우울증의다양한증세를분담하고있음을나타낸다 (Dranovsky et al., 2006; Duman et al., 2006). 예를들어 prefrontal/cingulated cortex, hippocampus, striatum, amygdale, thalamus, hypothalamus 등의뇌영역에서기능적변화가우울증발현과상관관계를보이고이는전통적인해부학적연구결과와많이일치하는결과로서이들뇌영역들이우울증연구의주요부위가된다 (Nestler et al., 2006). 우울증 episode가많은경우다양한형태의 stress 가있는정황에서일어난다는점은비록 stress가우울증을일으키는충분조건으로작용하지않을지라도우울증발현에중요한역할을할수있다는것이다. 생체에가해지는수많은 stress 자극은혈중과뇌조직각부위에서 DA를포함한 neurotransmitter의대사변화를유발하는것으로알려져있는데 (Murrin et al., 1975; LeBlanc et al., 1979; Dimsdale et al., 1980) 특히 stress에의한정서변화와관련된 mesocortical tract와 mesolimbic tract에관한연구가많이진행되어외부자극시이들조직에서 DA대사가항진되어있음이보고되었으며 (Murrin et al., 1975; Thirry et al., 1976) dopamine reuptake도증가된다고한다 (Gilad et al., 1986). 또한 serotonin 의합성과분비가증가되었음을보고하였다 (Kennet et al., 1987). 그러나이러한 stress에의한 neurotransmitter의대사변화는종간또는조직간의변화가다양하고 stress의종류와시간에따라많은차이를보임으로써 (Cabib et al., 1988), 특히 stress 자극시자주관찰되는운동성저하와관련될것으로생각되는여러 neurotransmitter의변화에대하여는아직많은연구가진행되어야할것으로생각된다. 398
김지현외 6 인 : 연자육의항우울효과에대한뇌부위별신경전달물질의변화 본연구의결과를볼때연자육의 NS37 추출물투여시 DA의함량증가및대사속도감소는이전연구 (Lee KS et al., 2008) 에서관찰되었던강제수영검사에서부동자세감소와등반시간증가등운동성의증가를뒷받침할수있는근거가될것으로생각된다. 따라서연자육의항우울효과는 NS37추출물이 DA계를주요매개체로그기능이발현되는것이라할수있다. 하지만운동성과직접적으로관련된 striatum에서의 DA 변화에대해서살펴보지못하였고, 등반자세의증가는 NE의증가를의미하는것으로알려져있으므로 (Deussing. 2006), 본연구에서살펴본결과외에다양한뇌부위별 neurotransmitters의변화및관련성에대한추가연구가진행되어야할것이다. 참고문헌 Barber J, Tomer R, Sroka H et al. S (1985) Dose unilateral dopamine deficit contribute to depression? Psychiatry Res. 15:17-24. Borsini F, Cesana R, Vidi A et al. (1991) Evidence that imipramine activities 5-HT1C receptor function. Eur. J. Pharmacol. 203:359-363. Borsini F, Pulvirent L, Samanin R (1985) Evidence of dopamine involvement in the effect of repeated treatment with various antidepressants in the behavioral despair test in rats. Eur. J. Pharmacol. 110:253-256. Brown AS, Gershon S (1993) Dopamine and depression. J. Neural. Transm. Gen. Sect. 91:75-109. Bunney WE Jr, Davis JM (1965) Norepinephrine in depressive reactions. A review. Arch. Gen. Psychiatry 13:483-494. Cabib S, Kempf E, Oliverio A et al. (1988) Effects of immobilization stress on dopamine and its metabolites in different brain areas of the mouse: role of genotype and stress duration. Brain Res. 441:153-160. Cervo L, Grignaschi G, Nowakowska et al. (1989) 1-(3-Trifluoromethylphenyl) piperazine (TEMPP) in the ventral tegmental area reduces the effect of desipramine in the forced swimming test in rats: Possible role of serotonin receptors. Eur J. Pharmacol. 171:119-125. Chojnacka Wojeik E, Tatarczynsk E, Golembiowska K et al. (1991) Involvement of 5-HT1A receptors in the antidepressant-like activity of gepirone in the forced swimming test in rats. Neuropharmacology 30:711-717. Coppen A (1967) The biochemistry of affective disorders. Br. J. Psychiatry 113:1237-1264. Deussing Jan M (2006) Animal model of depression. Drug discovery today: disease model 3. Dimsdale JE, Moss J (1980) Short-term catecholamine responses to psychological stress. Psychosomatic Med. 42:493-497. Drannovsky A, Hen R (2006) Hippocampal neurogenesis: regulation by stress and antidepressants. Biol. Psychiatry 59:1136-1143. Duman RS, Moteggia LM (2006) A neurotrophic model for stress-related mood disorders. Biol. Psychiatry 57:1116-1127. Duterte-Boucher D, Leclere JF, Panissaud C et al. (1988) Acute effects of direct dopamine agonists in the mouse behavioral despair test. Eur. J. Pharmacol. 154:185-190. Fuller RW (1995) Serotonin uptake inhibitors: uses in clinical therapy and in laboratory research. Prog. Drug. Res. 45:167-204. Gilad GM, Gilad VH, Rabey JM (1986) Dopaminergic modulation of the septohippocampal cholinergic system activity under stress. Life Sciences 39:2387-2393. Jang CG, Kang MK, Cho JH et al. (2004) Nelumbinis Semen reverses a decrease in 5-HI1A receptor binding induced by chronic mild stress, a depression-like symptom. Arch. Pharm. Res. 27:1065-1072. Kang MK, Pyun KH, Jang CG et al. (2005) Nelumbinis Semen reverses a decrease in hippocampal 5-HT release induced by chronic mild stress in rats. J. Pharmacy. & Pharm 57:651-656. Kennet GA, Dickinson SL, Curzon G (1985) Enhancement of some 5-HT dependent bahavioral reponses following repreated immobilization in rats. Brain. Res. 330:252-263. LeBlanc J, Cote J, Jobin M et al. (1979) Plasma catecholamines and cardiovascular responses to cold and mental activity. J. Appl. Physol. 47:1207-1211. Lee JH, Chung DK (1998) Effects of the Soyosan and Chyengansoyosan on the regional brain monoamines contents of cold swimming stressed mice. J. Orient. Neuropsychiatry 9:1-18. Lee KS, Cho CW, Kang M et al. (2008) The antidepressant effect of Nelumbinis semen on forced swimming test in the rat. The Korean Journal of Stress Research 13:99-106. Linde K, Berner MM, Kriston L (2008) St John's wort for major depression. Cochrane Database Syst Rev. 4:CD000448. Murrin L, Roth RH (1975) Dopaminergic neurons: Effects of electrical stimulation on dopamine biosynthesis. Mol. Pharmacol. 12:436-475. Nestler EJ, Barrot M, DiLeone RJ et al. (2002) Neurobiology of depression. Neuron 34:13-25. Nestler EJ, Carlezon WA Jr. (2006) The mesolimbic dopamine reward circuit in depression. Biol Psychiatry 59:1151-1159. Rihmer Z, Arato M (1990) Serum dopamine-beta-hydroxylase activity in psychotic and nonpsychotic major depression: The importance of distinguishing between unipolar and bipolar disorder. Acta Psychiatr. Scand. 82:93-94. Roy A, Karoum F, Pollack S (1992) Marked reduction in indexes of dopamine metabolism among patients with depression who 399
스트레스硏究 : 제 16 권제 4 호 2008 attempt suicide. Arch. Gen. Psychiatry 49:447-450. Sampson D, Willner P, Muscar R (1991) Reversal of antidepressant action by dopamine antagonist in an animal model of depression. Psychopharmacology 104:491-495. Tuirry AM, Tassin JP, Blanc G et al. (1976) Selective activation of the mesocortical DA system by stress. Nature. 263:242-244. Wolfe N, Katz DI, Albert ML et al. (1990) Neuropsychological profile linked to low dopamine in Alzheimer's disease, major depression, and Parkinson's disease. J. Neurol. Neurosurg Psychiatry 53:915-917. = 국문초록 = 선행연구에서연자육추출물중강제수영검사를통해항우울효과를검증하여가장뛰어난결과를보인 NS37 (n-buoh 추출물의추가분획물, H 2O:MeOH=3:7) 의항우울효과기전을규명하고자, HPLC-ECD 시스템을이용하여뇌의우울증신경회로로알려진대뇌피질 - 시상하부 - 해마의부위별세로토닌과도파민의함량을조사하고이들대사산물의측정및대사속도를검토하였다. 연구결과 NS37-10 (10 mg/kg) 군에서대뇌피질, 시상하부, 해마에서도파민의유의한증가및시상하부, 해마에서도파민교체율의유의한감소를나타냄으로써항우울효과를확인하였다. 중심단어 : 연자육, 항우울효과, HPLC, 세로토닌, 도파민 400