• 약학회지
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• 제52권6호
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• pp.419-425
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• 2008

A qualitative and quantitative analytical method was developed for detection of methamphetamine (MA) and its main metabolite amphetamine (AM) in oral fluid. Oral fluids of eleven drug abusers were provided by Police, specimens were collected by stimulation with a cotton swab treated with 20 mg of citric acid ($Salivette^{(R)}$; Sarstedt, USA). As the preliminary test, oral fluid samples were screened for amphetamines by Fluorescence Polarization Immunoassay (TDxFLx, Abbott Co.). Extraction for MA was performed using solid-phase extraction (SPE) by $RapidTrace^{TM}$ (Zymark, USA) with mixed mode cation exchange cartridge, CLEAN $SCREEN^{(R)}$ (130 mg/3 ml, UCT) after dilution with phosphate buffer. Samples were evaporated and derivatized by pentafluoropropionic acid anhydride (PFPA). Quantitation of MA and AM was performed by gas chromatography-mass spectrometry (GC-MS) using selective ion monitoring (SIM), the quantitation ions were m/z 204 (MA), 208 (MA-$D_5$), 190 (AM) and 194 (AM-$D_5$). The selectivity, linearity of calibration, limit of detection (LOD) and quantification (LOQ) within- and between day precision, accuracy and recoveries were examined as parts of the method validation. All oral fluid samples gave positive results to immunoassay for MA (cut-off level, 50 ng/ml as d-amphetamine). Concentrations of MA and AM by GC-MS in eleven samples were ranged 104.2${\sim}$4603.3 ng/ml and 32.4${\sim}$268.6 ng/ml, respectively. Extracted calibration curves of MA and AM were linear over the two concentration range of 1${\sim}$100 and 50${\sim}$1000 ng/ml with correlation coefficient of above 0.999. LOQ of MA and AM was 1 and 3 ng/ml, respectively. The intraand inter-day run precisions (CV) for MA and AM were less than 10%, and the accuracies (bias) for MA and AM were also less than 10% at the two different concentrations 5 and 100 ng/ml at low calibration range, 50 and 1000 ng/ml at high calibration range. The absolute recoveries of MA and AM at low and high calibration ranges were more than 82% and 75%, respectively. In this study the qualitative and quantitative analytical method of MA in oral fluid was established. Oral fluid testing may detect drug use in past hours because of its shorter detection window than urine, and be useful in post-accident situations. So oral fluids will be most useful for testing drug abuse in the driving under the influence of drug (DUID) as the alternative specimens of urine.

• 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.

• 대한약리학회지
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• 제20권2호
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• pp.35-47
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• 1984

실험적으로 L:D, 12:12또는 D:D, 12:12에 적응시킨 흰쥐의 간뇌에서 beta-endorphin함량의 일중변동과 이에 대한 수종 중추성 약물의 효과를 검토하고 beta-endorphin함량의 생리 및 약리학적 의의를 추구코저 $^3H-morphine$ binding을 측정하여 다음과 같은 성적을 얻었다. 1) L:D, 12:12 cycle에 적응시킨 대조군에서 뇌내 beta-endorphin함량은 06 : 00시 에 최고에 달하고 18 : 00시에 최저인 대단히 유의한 일중변동을 일으켰고 24시간 평균 beta-endorphin함량은 $46.7{\pm}3.6$ fmole/mg protein 이었다. 2) D:D, 12:12에 적응시친 표본에서 beta-endorphin 함량은 대조군에서와는 달리 14 : 00시에 최고에 이르고 02 : 00시에 최저에 이르는 일중변동을 보였으며 24시간 평균 beta-endorphin함량도 대조군에 비하여 현저히 감소되었다. 3) 뇌내 beta-endorphin함량 일중별동은 reserpine. pargyline, imipramine, amphetamine. 또는 chlorpromazine처리로써 변동되었다. 4) 대조군에서 $^3H-morphine$ binding은 22 : 00시에 최고에 달하는 일중변동을 보였으며, 지속적인 암적응군에서의 binding은 02 : 00최고, 그리고 14 : 00시에 최하에 이르는 일중변동을 보였다. 5) 대조군과 지속적인 암적응군에서 함량과 maximum $^3H-morphine$ binding간에는 유의한 역상관관계가 있었다. 이상의 실험성적으로 흰쥐 뇌내 beta-endorphin함량과 수용체는 유의란 일중변동을 일으키고 opiate receptor binding의 일중변동은 beta-·endorphin함량변동에 따른 receptor regulation에 의함을 시사하며 또한 지속적인 암적응을 포함하여 수종 중추에 작용하는 약물들이 endorphin함량 일중변동을 변동시킬 수 있음을 보여 준다.

• 대한약리학회지
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• 제20권2호
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• pp.23-34
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• 1984

실험적으로 명암주기 또는 암주기에 적응시킨 백서의 간뇌 homogenate에서 opiate receptor의 일중변동 유무를 검토하고, circadian rhythm에 영향을 미치는 것으로 알려진 수종 중추작용약물의 영향을 보고 저 7 group에서 4시간 간격으로 1일 6회 maximum $^3H-morphine$ binding을 측정하여 다음과 같은 성적을 얻었다. 1) L : D, 12 : 12에 적응시킨 대조군에서 maximum $^3H-morphine$ binding은 22시에 최고에 달하는 매우 유의한 일중변동을 보였고, 24시간 평균 $^3H-morphine$ binding치는 $0.45{\pm}0.03\;pmole/mg Protein이었다. 2) 지속적인 암적응을 시킨 D : D, 12 :12군에서 $^3H-morphine$ binding의 일중변동은 14시에 최하, 그리고 2시에 최고의 binding치를 보이는 만상성의 일주변동은 보였으며 대조군의 곡선과는 그 shape, Phase, 진폭, 최고 또는 최하 binding시기 및 24시간평균 opiate수용체의 수에 현저한 차이가 있었다. 3) 지속적인 암적응, reserpine, pargyline, imipramine, amphetamine 또는 chlorpromazine 처리는 opiate receptor의 일중변동을 변화시켰다. 4) 그러나 전 실험군에서 Kd치는 변동되지 않았다. 이상 실험성적으로 백서뇌내 opiate수용체의 일중변동은 수용체의 질적변동이 아닌 수적인변동에 의하고, 지속적인 암적응이나 circadian rhythm을 변동시키는 중추성 약물들이 opiate receptor의 일중변동을 변화시킬 수 있으며, 또한 동통을 포함하여 제종 진통제의 효과가 일중변동을 일으키는 것은 opiate receptor의 일중변동과 일정한 관계가 있을 것으로 추측하였다.

• 한국가축번식학회지
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• 제25권4호
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• pp.389-397
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• 2001

CART는 leptin에 의해 조절되는 포식인자이며 섭식과 운동 습성에 관계된 것으로 알려져 있다. 사람의 CART Leu34Phe 돌연변이는 비만의 표현형을 나타내었지만, 생쥐의 CART 돌연변이는 일반사료의 섭취 후 급격한 체중증가를 나타내지는 않았다 생체 내 신경세포에서 CART의 역할을 확인하기 위한 새로운 형질전환 모델을 확립하고자 분화하는 신경세포의 유전자 발현을 조절하는 NF-L promoter와 CART의 재조합 발현 벡터를 구축하였다. 형질전환 생쥐는 유전자 미세 주입법에 의하여 생산되었으며, PCR과 Southern blot의 방법으로 확인하였다. 이러한 형질전환 생쥐에서 CART의 과 발현을 수정 후 13.5일째 초기 배아와 생후 6주째 형질전환 생쥐의 뇌와 척수에서 확인하였다. 본 연구의 결과는 섭식 관련 유전자들이 상호 연관된 섭식행동에서 CART의 역할을 연구하는데 모델 동물로써 이용할 수 있을 것으로 사료된다.

• 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 $D_1R$.

• 대한약리학회지
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• 제21권2호
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• pp.90-98
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• 1985

새앙쥐 강제수영 실험에서 부동자세 시간의 단축은 향주도성 행동의 항진이라는 본능적 충동의 유발이라는 가정 아래 중추 noradrenaline neuron에 있어서 ${\alpha}_1$- 및 ${\alpha}_2$-adrenoceptor의 역할과 관련지어 이 실험을 행하였고 다음과 같은 결론을 얻었다. 1. 새앙쥐를 이용한 강제수영 실험에서 부동자세 시간은 clonidine 및 guanabenz 등과 같은 ${\alpha}_2$-agonists에 의하여 용량에 의존하여 단축되었다. B-HT 933 및 oxymetazoline은 용량에 의존하지 않으나 단축시켰다. xylazine에의하여는 오히려 증가되었다. 2.${\alpha}_1$-Agonists 인 cirazoline, amidephrine 및 methoxamine은 부동자세 시간에 일관성 있는 영향을 미치지 아니하였다. 3. Clonidine과guanabenz에 의한 부동자세 시간의 단축은 ${\alpha}_2$-antagonists, yohimbine, idazoxan 및 phentolamine 전처치로 봉쇄되었으나 ${\alpha}_1$-antagonists, prazosin 및 corynanthine에 의하여는 영향을 받지 아니하였다. 4, d-Amphetamine 투여시 부동자세 시간은 용량에 비례하여 단축되었고, 이러한 단축효과는 yohimbine에 의하여는 길항되었으나 prazosin에 의하여는 영향을 받지 아니하였다. 5. ${\alpha}$-methyl-p-tyrosine 이나 reserpine 또는 두 약물을 동시에 전처치 하였을때 clonidine에 의한 부동자세 시간의 단축은 영향을 받지 아니하였다. 6. Desipramine 및 imipramine 같은 항우울제를 장기처치 또는 장기간 저기충격 요법을 가한 새앙쥐에서도 clonidine의 효과는 영향을 받지 아니하였다. 이상의 결과로 보아 새앙쥐의 강제수영 실험에서의 부동자세 시간의 변동은 중추내 noradrenergic neuron의 postsynaptic ${\alpha}_2$-adrenoceptor와 밀접한 관련이 있다고 시사되며 이러한 ${\alpha}_2$-agonists에 의하여 항진되는 escape-directed behavior는 자기보호를 위한 일종의 충동의 유발로 인한 행동으로 사료된다.

• Biomolecules & Therapeutics
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• 제12권2호
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• pp.101-107
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• 2004

Repeated administration of psychostimulants such as amphetamines increases locomotor activity in rodents. These drugs, including methamphetamine, enhance dopaminergic neurotransmission and result in hyper-locomotion and behavioral sensitization. It is well known that the existence of a complex balance between the cholinergic and dopaminergic systems in the central nervous system. Thus, behavioral sensitization by methamphetamine may be related to the expression of the M1 muscarinic acetylcholine receptors gene. The present study investigated the changes of M1R mRNA in hyperlocomotor activity and behavioral sensitization by methamphetamine (2 mg/kg) in mice. Our results showed that M1R mRNA expression was increased in the frontal cortex and the hippocampus region (the CA2 region) in the acute methamphetamine administered group compared to the saline administered group. In the chronic group, M1R mRNA expression was increased in the frontal cortex ill1d the hippocampus regions (CA2 and DG regions) in melt1amphetamine administered group compared to saline control group. These results indicate that acute or chronic treatment of mathamphetamine leads to the region-specific changes in mRNA expression levels of M1R. Therefore, Therefore, the present result suggests that M1R may play a role in modulating of methamphetamine-induced behavioral sensitization in mice.

• 대한약리학회지
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• 제5권2호
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• pp.87-92
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• 1969

The effects of dephenylhydantoin, strychnine, coramine, d-amphetamine and chlorpromazine on $QO_2$ and non-inulin space $Na^+$, $K^+$ concentration of rat cerebral cortical slices incubated in pH 7.4 glycylglycine glucose saline was investigated. In general, there are decreased non-inulin space $Na^+$ concentration or increased non-inulin space $K^+$ concentration or both when the ratio of respiration to non-inulin space is greater than control group except in case of chlorpromazine $10^{-4}\;M$. And it is suggested that the ratio of respiration to non-inulin space is responsible more closely for the non-inulin space Na, K concentration than $QO_2$ expressed per tissue wet weight. Effects of diphenylhydantoin and several other agents on electrolytes and the electroshock seizure threshold are discussed.

• Journal of Microbiology and Biotechnology
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• 제32권9호
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• pp.1168-1177
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• 2022

Parkinson's disease (PD) is the second-most prevalent neurodegenerative disease and is characterized by dopaminergic neuronal death in the midbrain. Recently, the association between alterations in PD pathology and the gut microbiota has been explored. Microbiota-targeted interventions have been suggested as a novel therapeutic approach for PD. Agathobaculum butyriciproducens SR79^T (SR79) is an anaerobic bacterium. Previously, we showed that SR79 treatment induced cognitive improvement and reduced Alzheimer's disease pathologies in a mouse model. In this study, we hypothesized that SR79 treatment may have beneficial effects on PD pathology. To investigate the therapeutic effects of SR79 on PD, 6-hydroxydopamine (6-OHDA)-induced mouse models were used. D-Amphetamine sulfate (d-AMPH)-induced behavioral rotations and dopaminergic cell death were analyzed in unilateral 6-OHDA-lesioned mice. Treatment with SR79 significantly decreased ipsilateral rotations induced by d-AMPH. Moreover, SR79 treatment markedly activated the AKT/GSK3β signaling pathway in the striatum. In addition, SR79 treatment affected the Nrf2/ARE signaling pathway and its downstream target genes in the striatum of 6-OHDA-lesioned mice. Our findings suggest a protective role of SR79 in 6-OHDA-induced toxicity by regulating the AKT/Nrf2/ARE signaling pathway and astrocyte activation. Thus, SR79 may be a potential microbe-based intervention and therapeutic strategy for PD.