KISEP Original Article 백서에서알코올장기투여가도파민과세로토닌신경전달계에미치는영향 박성우 1 이유철 2 하은경 1 김영경 1 이용규 3 정도운 4 류정환 4 공보금 1 김영훈 1 인제대학교정신과학교실및백인제기념임상의학연구소, 1 동래병

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1 KISEP Original Article 백서에서알코올장기투여가도파민과세로토닌신경전달계에미치는영향 박성우 1 이유철 2 하은경 1 김영경 1 이용규 3 정도운 4 류정환 4 공보금 1 김영훈 1 인제대학교정신과학교실및백인제기념임상의학연구소, 1 동래병원, 2 동서대학교식품생명공학과, 3 마산동서병원 4 ABSTRACT Effects of Chronic Alcohol Exposure on Dopaminergic and Serotonergic Neurotransmission System in Rat Sung-Woo Park, PhD, 1 Yoo-Chul Lee, MD, 2 Eun-Kyung Ha, MD, 1 Young-Kyung Kim, MS, 1 Yong-Kyu Lee, PhD, 3 Do-Oun Chung, MD, 4 Jeoung-Whan Ryu, MD, 4 Bo-Geum Kong, MD 1 and Young-Hoon Kim, PhD, MD 1 1 Department of Neuropsychiatry, School of Medicine and Paik Institute for Clinical Research, Inje University, Busan, 2 Dongrae Hospital, Busan, 3 Department of Food and Biotechnology, Dongseo University, Busan, 4 Dongsuh Hospital, Masan, Korea Objective:This study investigates the effects of chronic alcohol exposure on rat brain THmRNA expression, TH (tyrosine hydroxylase) acitivity, and TPH (tryptophan hydroxylase) activity which are important in synthesis of dopamine and serotonin and other components of both the dopaminergic and serotonergic systems of the rat brain. Methods:Rats were fed a liquid diet containing alcohol for 4 weeks. We investigated effects of chronic alcohol exposure on dopaminergic systems as follows. We evaluated expression of THmRNA in LC, VTA and substantia nigra by using in-situ hybridization and measured activity of TH by using immunoassay. We used HPLC for simultaneous measurement of dopamine, DOPAC and HVA in the cerebral cortex, striatum, hypothalamus, hippocampus, mid brain, hind brain, and cerebellum. Also we investigated serotonergic systems as follows. We evaluated expression of TH mrna in the dorsal raphe nucleus by using radioprobe and measured the activity of TPH by using enzyme immunoassay. We used HPLC for simultaneous measurement of 5-HT and 5-HIAA in the cerebral cortex, striatum, hypothalamus, hippocampus, mid brain, hind brain, and cerebellum. Results:Alcohol exposure for 4 weeks increased the expression of TH mrna in the ventral tegmental area and the locus ceruleus but not in the substantia nigra. The 4 weeks of alcohol exposure did not cause significant changes in levels of dopamine and metabolites in the different areas of the brain, nor was it associated with changes in the maximal binding and affinity (Kd) of anterior striatal dopamine D2 receptor. Alcohol exposure for 4 weeks had no effect on the expression of TPH mrna or on the activity of TPH in the dorsal raphe nucleus and the hypothalamus. Conclusion:We reported at first that chronic alcohol exposure could increase TH mrna in the locus ceruleus. In a previous study of acute alcohol treatment, there is increase of dopamine metabolism but in this study, we did not observe any changes in dopamine metabolism in the different areas of the brain. Also we did not see any significant changes in the syn- 교신저자 81

2 thesis and metabolism of serotonin after 4 weeks of chronic alcohol exposure compared with control. Therefore, synthesis and metabolism of serotonin was affected in the acute phase. And, as previous reports have suggested, any changes caused by alcohol returned to previous levels via adaptation and regulatory mechanisms. (Korean J Psychopharmacol 2005;16(1):81-92) KEY WORDS:Alcohol Dopamine Serotonin TH (tyrosine hydroxylase) TPH (tryptophan hydroxylase). 서론 연구대상및방법 1. 연구대상 Korean J Psychopharmacol 2005;16(1):81-92

3 2. 연구방법 - 1) 알코올장기투여방법 2) 실험방법 - in vitro

4 in vitro Korean J Psychopharmacol 2005;16(1):81-92

- - - - - 3) 자료분석 연구결과 1. 장기알코올투여가도파민신경전달계에미치는영향 1) TH mrna 발현에미치는영향 Table 1.

ethanol diet-treated group N4 Region Control group N4 Ethanol group N4 p value Locus ceruleus 0.

5 ) 자료분석 연구결과 1. 장기알코올투여가도파민신경전달계에미치는영향 1) TH mrna 발현에미치는영향 Table 1. Tyrosine hydroxylase mrna level OD in the different areas of the brain of control group N4 and ethanol diet-treated group N4 Region Control group N4 Ethanol group N4 p value Locus ceruleus * Ventral tegmental area * Substantia nigra * ValuesMeanSD, p0.05, ODoptical density Locus ceruleus Ventral tegmental area Substantia nigra Figure 1. Tyrosine hydroxylase mrna expressed in locus ceruleus, ventral tegmental area, and substantia nigra. 85

6 알코올 장기투여의 영향 A B Figure 2. The in situ hybridization mrna expression of tyrosine hydroxylase (TH) in the locus ceruleus (LC, indicated by arrowheads). Alcohol treatment for 4weeks increased TH signal in the LC. A and B represent control and alcohol-treated groups, respectively. 86 Korean J Psychopharmacol 2005;16(1):81-92

7 2) TH 활성도에미치는영향 3) 도파민및그대사물의농도에미치는영향 - 4) 전선조체 (anterior striatum) 에서 H-spiperone 결합에미치는영향 - 2. 장기알코올투여가세로토닌신경전달계에미치는영향 1) TPH mrna 발현에미치는영향 2) TPH 활성도에미치는영향 Table 2. Tyrosine hydroxylase activity nmol/mg/min in the different areas of the brain of control and ethanol diet-treated groups Region Control group N6 Ethanol group N6 Locus ceruleus NS Midbrain NS Caudate nucleus NS ValuesMeanSD, NSnot significant p Table 3. Mean concentrations of dopamine, serotonin and their metabolites in the different areas of the brain ng/ul of control group N10 and ethanol diet-treated group N10 Cerebral cortex Striatum Hypothalamus Hippocampus Midbrain Region Group Dopamine DOPAC HVA 5-HT 5-HIAA Control Ethanol p NS NS NS NS NS Control Ethanol p NS NS NS NS NS Control Ethanol p NS NS NS NS NS Control Ethanol p NS NS NS NS NS Control Ethanol p NS NS NS NS NS Control Hindbrain Ethanol p NS NS NS NS NS ValuesMeanSD, NSnot significant 87

8 Table 4. 3 H-spiperone binding to anterior striatum of control and ethanol diet-treated groups Bmax fmol/mg protein Control group N ) 세로토닌및그대사물의농도에미치는영향 - - 고 찰 Ethanol group N4 p NS Kd nm NS ValuesMeanSD, NSnot significant Table 5. Tryptophan hydroxylase mrna levels OD in dorsal raphe nucleus of control and ethanol diet-treated groups N Control group N5 Ethanol group N MeanSD NS ValuesMeanSD, NSnot significant Table 6. Tryptophan hydroxylase activity nmol/mg/min in the different areas of the brain of control N6 and ethanol diet-treated groups N6 Region Control group N6 Ethanol group N6 Raphe nucleus NS Hypothalamus NS ValuesMeanSD, NSnot significant p p 1. 알코올이도파민에미치는영향 88 Korean J Psychopharmacol 2005;16(1):81-92

9 - 2. 알코올이세로토닌에미치는영향 89

10 - in vitro in vivo - 90 Korean J Psychopharmacol 2005;16(1):81-92

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