• 약학회지
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• 제43권6호
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• pp.743-750
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• 1999

Human and bovine dopamine transporters (DAT) demonstrate discrete functional differences in the dopamine (DA) transport and cocaine binding. The functional analyses on the chimeras of human and bovine DAT have revealed that the region from the $133^{rd}{\;}to{\;}186^{th}$ residue(encompassing the $3^{rd}$ trans-membrane domain (TM) is responsible for the substrate transport and cocaine binding. The present studies have been done to find out the specific amino acid(s) which is essential for the binding of cocaine to DAT by interchanging the amino acids in that region between human and bovine DAT. When isoleucine, the $152^{nd}$ residue of chimera B3 (bovine DAT sequence) was transformed back to valine, the human DAT residue at the identical position, the cocaine binding was remarkably recovered to 98% of the human DAT values. In addition, the cocaine binding of the human DAT was decreased by 57% by substituting isoleucine for valine at position 152. When isoleucine at position 152 of the chimera B3 was converted to the other amino acids to provide an possible molecular basis for the functional role of the $152^{nd}$ residue, only the conversion to alanine among acids tested significantly the cocaine by 34%, but these effect were not as much as those by the conversion to valine. In conclusion, valine at position 152 is a crucial amino acid for the interaction of cocaine to the DAT.

• Toxicological Research
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• 제6권1호
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• pp.75-88
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• 1990

The characteristics of dopamine uptake, D-1 and D-2 receptors after acute and subacute cocaine administration were determind in striatum from WKY and SHR. Cocaine was administered either acutely (40 mg/kg, s.c.) or twice daily (20 mg/kg, s.c.) for 3 and 7 days in 9-wk old WKY and SHR. Rats were sacrificed 30 min, 2 or 24 h after the single injection and 18 h after the last administration to the subacutely treated group. The changes in dopamine uptake, dopamine uptake sites, D-1 and D-2 receptors were determined using $(^3H)$dopamine, $(^3H)$-GBR-12935, $(^3H)$SCH-23390 and $(^3H)$sulpiride, respectively. In acutely treated rats, significant increases in $V_{max}$of dopamine uptake were observed 30 min after the cocanine injection in both strains without changes in $K_m$ values. The in vitro $IC_{50}$for cocaine was significantly decreased 30 min in WKY and 2 h in SHR. However, that for in vitro GBR-12909 was significantly increased 30 min and 2 h in both strains. Also densities of $(^3H)$-GBR-12935 binding sites were significantly increased 30 min and 2 h without changes in their $K_d$. Significant increases in D-2 receptor density were observed 30 min, 2 or 24 h after acute injection in both strains without changes in their affinities. The density of D-1 receptor was significantly decreased 30 min after the injection in WKY, but not in SHR. In subacutely treated rats, a significant increase in $K_m$ of dopamine uptake was observed in 7-day treated SHR. The in vitro $IC_{50}$fot GBR-12909 was significantly increased in 3-day treated WKY. The density of D-1 receptors was significantly increased in 3- and 7-day treated WKY, but not in SHR. The affinity of both binding sites remained unchanged. The results suggest that cocanine administration alters dopamine uptake, characteristics of dopamine uptake sites and dopamine receptor binding characteristics in rat brain. Furthermore, D-1 and D-2 dopamine receptors appear to be differently regulated.

• 동의생리병리학회지
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• 제26권3호
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• pp.281-286
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• 2012

The present study was designed to investigate the effect of Rehmannia glutinosa on phosphorylation of extracellular signal-regulated kinase(ERK) and cAMP response element-binding protein(CREB) in the acute cocaine-treated rats. Rats orally received vehicle or extract of Rehmannia glutinosa 1 h prior to saline (1 ml/kg, i.p.) or cocaine hydrochloride (20 mg/kg, i.p.) treatment. Rats were sacrificed 15 min after a single intraperitoneal injection of saline or cocaine. Rehmannia glutinosa at dose of 50 mg/kg significantly decreased phosphorylation of ERK, CREB and Elk-1 in the nucleus accumbens and striatum of the cocaine-treated rat brain by immunocytochemistry. These results suggest that Rehmannia glutinosa may contribute to the effects of cocaine on gene expression and on behaviors.

• The Korean Journal of Physiology and Pharmacology
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• 제19권2호
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• pp.89-97
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• 2015

The aim of this study was to investigate the involvement of dopaminergic receptors (DR) in behavioral sensitization, as measured by locomotor activity, and the over-expression of cocaine- and amphetamine-regulated transcript (CART) peptides after repeated administration of cocaine in mice. Repeated administrations of cocaine induced behavioral sensitization and CART over-expression in mice. The levels of striatal CART mRNA were significantly increased on the $3^{rd}$ day. CART peptides were over-expressed on the $5^{th}$ day in the striata of behaviorally sensitized mice. A higher proportion of $CART^+$ cells in the cocaine-treated mice were present in the nucleus accumbens (NAc) shell than in the dorsolateral (DL) part of caudate putamen (CP). The concomitant administration of both $D_1R$ and $D_2R$ antagonists, SCH 23390 ($D_1R$ selective) and raclopride ($D_2R$ selective), blocked cocaine induced-behavioral sensitization, CART over-expression, and cyclic adenosine 5'-monophosphate (cAMP)/ protein kinase A (PKA)/phospho-cAMP response element-binding protein (pCREB) signal pathways. SCH 23390 more predominantly inhibited the locomotor activity, CART over-expression, pCREB and PKA activity than raclopride. Cocaine induced-behavioral sensitization was also attenuated in the both $D_1R$ and $D_2R$ knockout (KO) mice, respectively. CART over-expression and activated cAMP/PKA/pCREB signal pathways were inhibited in the $D_1R$-KO mice, but not in the $D_2R$-KO mice. It is suggested that behavioral sensitization, CART over-expression and activated cAMP/PKA/pCREB signal pathways induced by repeated administration of cocaine could be more predominantly mediated by $D_1R$.

• Biomolecules & Therapeutics
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• 제19권1호
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• pp.45-51
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• 2011

These experiments were designed to use typical makers from behaviors and molecular basis in addicted animals of morphine and cocaine. Morphine has been widely abused with a high physical dependence liability. Morphine withdrawal activates the intracellular cAMP signaling pathway and further leads to changes in the expression of the cAMP response element binding protein (CREB), which may be important to the development and expression of morphine dependence. From these experiments, repeated morphine (10 mg/kg, twice per day for 7 days) developed physical dependence. Withdrawal signs were precipitated by naloxone and also increased the expression of the CREB. In addition, repeated exposure of cocaine (15 mg/kg) to mice develops locomotor sensitization and produced lasting behavioral sensitivity. Cocaine- and amphetamine-regulated transcript peptide (CART) peptide was up-regulated by repeated administration of cocaine in the striatum. Therefore, repeated morphine induced the development of physical dependence and increased pCREB. In addition, repeated cocaine induced locomotor sensitization and over-expressed CART peptide. In conclusion, the development of physical dependence and pCREB for morphine, and locomotor sensitization and CART peptide over-expression for cocaine would be useful markers to predict the abuse potential of opioid analgesics and pychostimulant drugs in animals, respectively.

• 약학회지
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• 제36권2호
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• pp.188-190
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• 1992

Two clones of monconal antibodies(Co-1 and Co-2) against BSA-benzoylecgonine(BSABE) were produced. Both monoclonal antibodies showed high binding affinity to BSA-BE. Observing from ELISA inhibition assay, Co-1 reacted only weakly with soluble benzoylecgonine, while Co-2 showed considerable reactivity with soluble benzoylecgonine.

• Archives of Pharmacal Research
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• 제23권4호
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• pp.360-366
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• 2000

The in vivo binding of dopamine (DA) radioligands to $D_2$receptors can be affected by competition with endogenous dopamine. In the present study, we used a brain slice preparation that provides more controlled conditions than in vivo preparations in order to examine the relationship between synaptic DA and the binding of [$^3H$] raclopride to $D_2$receptors. We also estimated the synaptic DA concentration in rat striatal slices by determining the changes in [$^3H$] raclopride binding. To correlate the changes in [$^3$H]raclopride binding with the concentration of synaptic DA, the kinetic parameters were determined. [$^3H$] Raclopride reached equilibrium binding conditions within two hours. The K value for DA in inhibiting [$^3$H]raclopride binding was about 2.2 nM. The increase in synaptic DA evoked by electrical stimulation decreased the striatal binding of [$^3H$] raclopride in a frequency-dependent manner. Increases in the DA concentration evoked by amphetamine (AMPH) or cocaine decreased [$^3H$] raclopride binding by 74% or 20%, respectively, corresponding to increases in the synaptic DA concentrations of 1.6 nM or 0.6 nM, respectively. Pargyline also decreased [$^3H$] raclopride binding by 36%corresponding at a concentration of 1.2 nM. In contrast, the depletion of synaptic DA by $\alpha$-methyl-p-tyrosine ($\alpha$-MpT) increased the specific binding of [$^3H$] raclopride by 43%when the DA concentration was decreased to 0.7 nM. The changes in the DA concentration at the synapse were responsible for the changes in the striatal binding of [$^3H$] raclopride. The values calculated in this study may therefore approximate the changes in the synaptic DA concentration in rat striatal slices following manipulation.

• Journal of Ginseng Research
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• 제32권1호
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• pp.1-7
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• 2008

Ginseng saponin has been shown to inhibit the development of dependence on morphine, cocaine, methamphetamine, but the antinarcotics effects of ginseng on nalbuphine remains still largely unknown. Ginseng administration attenuated the naloxone-induced jumping behavior on nalbuphine dependent mice. The development of morphine dependence was mediated through ${\mu}-opioid$ receptor, however, development of nalbuphine dependence was mediated through ${\kappa}-opioid$ receptor. However, it was found that the efficacy of analgesic antagonism of GTS was mediated through the serotonergic mechanism, not mediated through the opioid receptor. In addition, ginseng administration modulated cellular signal transduction in the brain. The increased NMDA receptor subunit (NR1, pNR1), phosphate extracellular signal regulated protein kinase (pERK), phosphate cAMP response element binding protein (pCREB) expression by nalbuphine was decreased by the administration of ginseng powder in cortex, hippocampus, striatum of rat brain. These results suggest that ginseng could be one of the targets of antinarcotic therapies to reduce the development of tolerance and dependence on nalbuphine as well as morphine.

• 대한핵의학회지
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• 제39권6호
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• pp.481-488
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• 2005

목적: 도파민운반체의 기능적 영상술은 도파민 신경계의 시냅스전 신경말단의 상태를 나타내 줄 수 있어, 기저신경절을 침범하는 파킨슨 병 등의 진단 및 중증도 판단에 이용될 수 있으며, 도파민운반체가 코카인이나 methamphetamine 등 의존성 약물의 목표 부위이므로 이러한 약물의 남용자의 평가에 이용될 수 있다. 본 연구는 한국에서 생산된 도파민운반체 영상용 방사성의약품을 사용한 뇌 SPECT영상을 이용하여 methamphetamine 남용자에서 나타나는 소견과 이의 임상적 의의를 알아보고자 하였다. 대상 및 방법: Methamphetamine 남용자 6명 (남용자군, 전부 남자, 평균연령: $32{\pm}6.8$) 및 정상대조군 4명(대조군, 남:여=3:1, 평균연령=$44.0{\pm}14.7$세)을 대상으로 하였다. 남용자군 6명 모두에서 뇌 MRI가 시행되었다. 남용자군에서는 정신과적 증상과 우울증 증상을 정량적으로 평가하였다. 도파민운반체 영상은 I-123 IPT 185 MBq을 정맥 주사한 후 2시간에 삼중헤드 감마카메라(Prism 3000, Picker, USA)를 이용하여 뇌 SPECT 영상을 얻었다. 정성적인 판독과 함께, 정량적 분석으로 기저신경절 부위에 관심영역을 그려 기저신경절의 방사능량을 구하였고, 배후 방사능량은 후두엽 부위에 관심영역을 그려 방사능량을 구하였다. 기저신경절과 배후 방사능량을 이용하여 기저신경절 도파민운반체 특이결합/비특이결합 비율(특이결합비) (기저신경절방사능량-배후방사능량/배후방사능량)을 구하였다. 각군의 도파민운반체 특이겹합비를 비교하였으며, 남용자군에서는 정신과적 증상(Brief Psychiatric Rating Scale: BPRS) 및 우울증 증상 정도(Hamilton Depression Rrating Scale : HAMD)와 도파민운반체 특이겹합비의 상관성을 알아보았다. 결과: 남용자군 6명 모두는 뇌 MRI상 특이 소견이 관찰되지 않았으나, 모든 환자는 정신과적 증상과 우울증 증상을 나타내었으며 BPRS 점수와 HAMD 점수는 서로 밀접한 상관성을 보여 주었다(r=1.0, p=0.005). 남용자군의 I-123 IPT 뇌 SPECT의 정성적분석에서 5명(83.3%)이 기저신경절의 전체적 섭취가 불균등하고 미측의 섭취가 감소되어 비정상으로 판독 되었으며, 1명은 정상으로 판독되었다. 남용자군은 대조군에 비하여 낮은 기저신경절 도파민운반체 특이겹합비를 나타내었다($2.38{\pm}0.20\;vs\;3.04{\pm}0.27$, p=0.000) 남용자군의 BPRS 점수와 HAMD 점수가 높을수록 낮은 기저신경절 특이겹합비를 나타내어 역의 상관관계를 보였다(r=-0.908, p=0.012, r=-0.924, p=0.009). 결론: I-123 IPT뇌 SPECT는 흑질-선조체 도파민신경계의 손상을 일으키는 중증의 methamphetamine 남용자의 평가에 이용될 수 있다고 생각된다.