• 한국응용곤충학회지
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• 제31권2호
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• pp.122-132
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• 1992

신규화합물의 in Vitro AChE 저해력을 평가하기 위한 저해시험법의 확립을 목적으로 실험을여 다음과 같은 결과를 얻었다. {TEX}$I_{50}${/TEX} 산출이 가능한 고정시간 저해시험(fixed time inhibition test)의 경우, DFP, DDVP 및 paraoxon과 같은 대표적 AChE 저해제의 저해력 평가에 적당한 저해시간은 10분 정도로 조사되었으며, 10분간의 저해조건하에서 ester group을 보유한 신규화합물의 저해여부 및 정도의 평가에 효과적인 시험농도는 10$\mu$M 수준으로 판정되었으나 다른 화합물의 경우에는 보다 높은 농도로부터 단계적 저해시험이 필요한 것으로 보였다. {TEX}$K_{d}${/TEX}, {TEX}$k_{3}${/TEX} 및 {TEX}$k_{i}${/TEX}와 같은 저해상수의 산출을 위한 경시적 저해시험(Progressive inhibition test)에 있어서, 본 실험의 장시간 저해(0.8~13.3분)와 낮은 저해제농도(1$\times${TEX}$10^{-9}${/TEX}~2$\times${TEX}$10^{-6}${/TEX}M)의 조건하에서는 극히 낮은 {TEX}$K_{d}${/TEX}(1.3$\times${TEX}$10^-{8}${/TEX}~5.6$\times${TEX}$10^{-7}${/TEX}M) 및 {TEX}$K_{3}${/TEX}(0.21~{TEX}$0.27min^{-1}${/TEX})가 관찰되었으나 저해제간의 저해독성 조사에는 충분한 타당성을 가지는 것으로 조사되었다. 또한 DFP, paraoxon 및 신규화합물인 KH501간의 현격한 저해력 차이는 {TEX}$K_{d}${/TEX}의 차이 즉, 저해제와 AChE간의 친화력 차이에 기인된 것으로 판명되었다. 따라서 AntiChE 스크리닝에 있어서 1차단계에서는 고정시간 저해법을 통하여 산출된 {TEX}$I_{50}${/TEX}를 바탕으로 하여 저해력 유무 및 정도를 파악하고, 1차단계에서 고도의 AChE 저해력이 인정된 신규 화합물을 대상으로 2차단계인 경시적 저해법을 이용하여 저해양상 및 특성을 조사하는 것이 합리적인 체계일 것으로 생각되어진다.

• 한국환경과학회지
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• 제17권12호
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• pp.1325-1330
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• 2008

This study was carried out with the detection for multiresidue of the carbamate pesticide such as carbaryl and cabofuran by enzyme-inhibition method. The check time for determination of acetylcholinesterase(AChE) activity was selected at 60 sec. The AChE activity in chicken brain determined by the Ellman's method was $162{\mu}$mol/min/g protein. $I_{50}$ for AChE by carbamate pesticide with wet kit was 0.169mg/L of carbaryl and 0.089mg/L of cabofuran, respectively. The incubation time for enzyme kit with substrate kit was 30min for determination of AChE activity. Enzyme kit with substrate kit was stable at $4^{\circ}C\;and\;25^{\circ}C$ for 5 days. Limit detection concentration of carbaryl with dry kit for AChE was 0.05mg/L. The dry kit such as wet kit applied Enzyme-Inhibition(EI) method with AChE was confirmed the multi residue method to detect the carbamate pesticides.

• 한국응용곤충학회지
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• 제31권1호
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• pp.1-6
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• 1992

점박이응애(Tetranychus urticae Koch)에 대한 독성시험방법을 비교하고, 감수성계통을 누대도태하여 in vivo에서 저항성 발달수준과 in vitro에서의 AChE insensitivity의 관계를 실험한 결과는 다음과 같다. 점박이응애의 살비제 저항성 수준결정의 독성시험방법중 slide dip법 (CV=8.7%)은 leaf dip법(CV=12.2%) 이나 leaf disc법(CV=13.6%)보다 변이가 적고 정확도가 높을 뿐만 아니라 처리방법이 간편하고 한가지 농도에 대하여 여러 계통을 동시에 처리할 수 있고 처리 후의 조건을 균일하게 유지시킬 수 있으며 기주식물 없이 시험이 가능하였다. topical application법(CV=8.1%) 역시 정확도는 높았으나 시험 조작이 어렵고 필요한 장비의 구입 비용이 더 들며 시간이 많이 소요되는 단점이 있었다. 두 유기인제에 의한 누대도태결과 22세대 carbophenothion 도태개체군은 동일약제에 156배의 저항성이 발달하였으며, ethion에 대하여 128배의 저항성 반응을 보였다. 24세대 ethion 도태개체군은 동일약제에 64.1배의 저항성 발달이 되었으며, carbophenothion에 대하여도 65배의 저항성 반응을 보였다. 도태개체군의 AChE 활성저해도를 보면 감수성계통에 비하여 carbophenotion 누대계통은 carbohenothion은 3.3배, ethion은 2.7배의 활성저해도를 보였다. 또한 ethion 도태개체군에 대해서는 ethion은 2.6배 carbophenothion은 3.0배의 활성저해도를 나타냈다. 위와 같이 in vivo에서의 저항성 발달 수준과 in vitro에서의 AChE 저해도와는 밀접한 관계가 있는 것으로 보아 유기인계 살비제에 대한 점박이응애의 저항성 발현에는 AChE의 insensitivity가 주요인으로 작용하는 것으로 생각된다.

• 셀메드
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• 제2권2호
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• pp.19.1-19.6
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• 2012

$Capparis$ $zeylanica$ Linn. a 'Rasayana' drug is used for its memory enhancing effects in the traditional Ayurvedic system of medicine. The aim of this study was to evaluate acetylcholinesterase (AChE) inhibitory and memory enhancing activities of $Capparis$ $zeylanica$ Linn. The$in-vitro$ and $ex-vivo$ models of AChE inhibitory activity were used along with Morris water maze test to study the effect on memory in rats. The anticholinesterase effect of methanolic and aqueous extracts of $Capparis$ $zeylanica$ was measured by spectrophotometric Ellman method at 0.1, 0.3, 1.0, 3.0, 10 and 30 mg/ml and brain monoamine oxidase (MAO-A and MAO-B) activity was assessed by Naoi's method. The results $in-vitro$ and $ex-vivo$ AChE assay revealed that methanolic and aqueous extracts of $Capparis$ $zeylanica$ inhibit AChE activity, whereas these extracts did not alter MAO activity at any concentration tested as compared to moclobemide and L-deprenyl. The results indicate that $Capparis$ $zeylanica$ improves scopolamine-induced memory deficits through inhibition of AChE activity, and not by direct MAO inhibition.

• 동의신경정신과학회지
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• 제28권2호
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• pp.123-136
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• 2017

Objectives: The objective of this study is to investigate the anti-amnesic effects of AR, Aucklandiae Radix, ground powder on scopolamine (Sco)-induced memory impairment in mice (C57BL/6) through its favorable acetylcholine (ACh) and acetylcholinesterase (AChE) activity, Choline acetyltransferase (ChAT) mRNA expression, and antioxidant effect. Methods: Six groups, a total of 20 intact or 100 Sco treated mice, were selected based on their body weights and were used in this study. Half of the mice in each group were used for the passive avoidance task test and the measurements of hippocampus ACh content, AChE activity and ChAT mRNA expression. The remaining half of the mice in each group were used for the Morris water maze test and cerebral antioxidant defense system measurement. Results: Marked decreases in step-through latency times in the passive avoidance task test and increases in escape latency times in the Morris water maze test were observed with decreases in the hippocampus ACh content and ChAT mRNA expression, and increases in the hippocampal AChE activities, as a result of Sco intraperitoneal treatment, in the present study. In addition, destruction of the cerebral cortex antioxidant defense systems was observed in Sco control mice as compared with intact vehicle control mice. However, 28 days of continuous oral pre-treatment with AR ground powder at doses of 400, 200 and 100 mg/kg markedly and dose-dependently inhibited the Sco treatment-related amnesia. Conclusions: The results prove that oral administration of AR ground powder reduces Sco-induced memory impairment. This is because it can preserve ACh, related to ChAT mRNA expression, cause AChE inhibition, and activate the cerebral antioxidant defense system.

• 동의신경정신과학회지
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• 제30권3호
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• pp.221-235
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• 2019

Objectives: The purpose of this study was to confirm the anti-amnesic effects of Bunsimgieum (BSGE) through its favorable acetylcholine (ACh) and, acetylcholinesterase (AChE) activity, choline acetyltransferase (ChAT) mRNA expressions, and antioxidant effect on scopolamine (Sco)-induced memory impairment in C57BL/6 mice. Methods: Six groups, a total of 20 intact or 100 Sco-induced mice were used in this study, based on their body weight. Half of each group underwent passive avoidance tests and the measurement of hippocampus AChE activity, ACh content, and ChAT mRNA expression, The remaining half of each group underwent a Morris water-maze test and antioxidant defense system measurement as well. Results: Significant reductions in the step-through latency times from the passive avoidance test and reductions in the escape latency times from the Morris water-maze test were observed with increases of hippocampal AChE activities and, reductions in ACh contents and ChAT mRNA expression in hippocampus, as a result of Sco intraperitoneal treatment, in this study. Additionally, the increases in cerebral cortical MDA levels and, reductions in GSH contents, SOD activities, and CAT activities were demonstrated in the Sco control mice compared with the intact vehicle control mice, respectively. However, 28 days of consecutive oral pre-treatment of BSGE hot water extracts of 400, 200, and 100 mg/kg, respectively, markedly and dose-dependently inhibited Sco treatment-related amnesia. Conclusions: The results demonstrate that the oral administration of BSGE hot water extracts reduces Sco-induced memory impairment, through preserving ACh, related to ChAT mRNA expressions, causes AChE inhibition, and enhances the cerebral antioxidant defense system.

• 동의신경정신과학회지
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• 제29권3호
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• pp.121-134
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• 2018

Objectives: The purpose of this in vivo study is to observe anti-amnesic effects of Yeongkyekamjotanggayonggolmoryo (YGYM), a novel mixed herbal prescription, Ossis Mastodi and Ostreae Testa added Yeongkyekamjo-tang, on scopolamine induced amnesia in C57BL/6 mice through acetylcholine (ACh) and acetylcholinesterase (AChE) activity, Choline acetyltransferase (ChAT) mRNA expression, and antioxidant effects. Methods: Six groups, total 20 intact or 100 Sco treated mice were used in this study after one week of acclimatization period. Half the animals were used for passive avoidance task tests and hippocampus ACh content, AChE activity, and ChAT mRNA expression were measured. The other half was subjected to an underwater maze test and then the cerebral cortex antioxidant defense system was measured. Results: In the passive avoidance experiment, there was significant decrease in residence time in the bright room and in the underwater maze test, escape latency to escape from the esophagus significantly increased compared with the normal control group. At the final sacrifice, ACh content and ChAT mRNA expression decreased, AChE activity increased, and cerebral cortical MDA increased GSH content, SOD and CAT activity in Sco control mice, as compared to intact vehicle control mice. However, these Sco treatment-related memory loss through AChE activation destroyed the cerebral cortex antioxidant defense system, and was inhibited dose-dependently by 28 days consecutive oral pretreatments of YGYM extracts 500, 250, 125 mg/kg. Conclusions: In the above results, YGYM extract that oral administration of YGYM extracts alleviates the antioxidant defense system, through preservation of ACh mediated by upregulation of ChAT mRNA expression, and increase of AChE inhibition and brain antioxidant defense systems.

• 농업생명과학연구
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• 제45권6호
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• pp.213-226
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• 2011

아세틸콜린 분해효소(acetylcholine esterase, AChE) 억제제는 아세틸콜린 함량을 높여 콜린성 neuron을 활성화함으로써 기억 능력의 개선 및 치매 개선을 가져와 현재 다양한 AChE 억제제들이 개발되어 사용되고 있다. 본 연구에서는 AChE에 대한 억제 활성을 갖는 천연물을 다양한 식물추출물 및 에센스오일로부터 탐색하였으며, 탐색한 추출물의 scopolamine으로 기억손상을 유발한 쥐의 기억력 개선 활성을 치매 치료제로 사용하고 있는 donepizil과 비교 분석하였다. 그 결과 자몽(Citrus paradisi) 유래의 에센스 오일이 AChE 억제 활성이 가장 높아 20 ug/ml의 농도로 처리하였을 때 90% 이상의 효소 억제 활성을 나타내었다. 수동회피 실험 결과, 자몽 유래의 에센스오일(100 mg/kg, p.o.)을 투여한 쥐는 치매 치료제로 사용하고 있는 donepizile (0.5 mg/kg)을 투여한 쥐와 유사한 latency time을 나타내어 인지기능이 개선되었다. 또한, 수중미로 시험 결과, 자몽 유래 에센스오일(100 mg/Kg, p.o.)을 투여한 쥐는 donepizile(0.5 mg/kg)을 투여한 쥐와 유사한 latency time을 나타내어 인지기능이 개선되었다. 이상의 결과로부터 자몽 유래 에센스오일은 매우 효과적으로 기억력을 개선하여 인지기능을 개선해 줄 수 있는 안전하고 효과적인 후보물질이라고 사료된다.

• 대한본초학회지
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• 제22권4호
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• pp.51-58
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• 2007

Objectives : Investigation of the memory and cognitive enhancing effect of HT008-1 in scopolamine induced amnesia mice. Methods : At 60 min before acquisition trials, HT008-1 (30, 100, 300 mg/kg p.o.) was administered, and 30 min later, mice were injected with scopolamin (1.0 mg/kg, i.p.). In the passive avoidance test, acquisition trials were carried out 30 min after a single scopolamine treatment. Retention trials were carried out 24h after acquisition trials. Y-maze test was carried out 30 min after a single scopolamine treatment. Spontaneous alternation behavior during an 8-min session was recorded. Inhibitory effects of HT008-1 (0.01, 0.1, 1.0 mg/ml) on AChE activity was measured. Result : HT008-1 ameliorated scopolamine-induced learning impairments and spatial cognitive function in passive avoidance and Y-maze test, respectively. Moreover HT008-1 showed a significant inhibitory effect on AChE activity. Discussion: This study presented that eMultiherb mixture HT008-1 enhanced learning memory and spatial cognitive function in scopolamine-induced amnesia mice. These results suggest that the effect of HT008-1 may be dependent on the inhibition of AChE activity.

• 동의신경정신과학회지
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• 제29권4호
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• pp.207-221
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• 2018

Objectives: The objective of this study was to observe the anti-amnesic effects of Yukmijihwang-tang (YMJHT), on the scopolamine (Sco)-induced memory impairment in C57BL/6 mice through its favorable acetylcholine (ACh). Also, to observe acetylcholinesterase (AChE) activity, Choline acetyltransferase (ChAT) mRNA expressions, and antioxidant effect. Methods: Six groups, with a total of 20 normal and 100 Sco treated mice were selected based on their body weights after 1 week of acclimatization, were used in this study as follows. Half of the mice in each group were used for passive avoidance task tests and hippocampus ACh content, AChE activity and ChAT mRNA expression measurement, and the remaining half in each group used for Morris water maze test and measurement of cerebral antioxidant defense system. Results: Amnesia due to AChE activations and destroyed cerebral cortex antioxidant defense systems were markedly and dose-dependently inhibited after 28 days of continuous oral pre-treatment with YMJHT 400, 200 and 100 mg/kg, respectively. The overall effects of YMJHT 400 mg/kg were similar to those of tacrine 10 mg/kg. Conclusions: Based on the results, it was established that oral administration of YMJHT favorably alleviates Sco-induced memory impairment, through preservation of ACh, mediated by up-regulation of ChAT mRNA expressions and related AChE inhibition and augmentation of cerebral antioxidant defense system, at least in a condition of this experiment. The overall effects of YMJHT 400 mg/kg were similar to those of tacrine 10 mg/kg.