• 제목/요약/키워드: Ellman's Test

검색결과 8건 처리시간 0.017초

하수오 지상부의 활성 성분 (Active Compounds from The Aerial Parts of Polygonum multiflorum Thunberg)

  • 김진표;김도훈;김기은;김진효;정성희;김성건;황완균
    • 약학회지
    • /
    • 제50권6호
    • /
    • pp.351-357
    • /
    • 2006
  • Polygoni Radix (Polygonaceae) has been used as renal disoder hematopoietic, menstrual irregularity and antiaging in Korean folk medicine. Specially aerial parts has been used for insomnia and sedative agent. In order to investigate the efficacy of antioxidant activity on these aerial parts, the bio-activity guided fraction and isolation of physiologically active substance were performed. H$_2$O, 30%, 60%, 100% MeOH were examined antioxidant activity by DPPH method. It was revealed that 30%, 60% MeOH fractions have significantly antioxidant activity. From 60% MeOH fraction, six known flavonoid glycosides (quercetin, quercetin 3-O-${\beta}$-D-galactopyranosyl-4'-O-glucopyrano side-3'-methyl ether, quercetin 3-O-${\alpha}$-L-rhannopyranoside, quercetin 3-O- ${\alpha}$-L-rhamnopyranoside-4'-methylethor, quorcetin 3-O-${\beta}$-D-galactopyranoside, quercetin 3-O-${\beta}$-D-galactopyranoside) and 3,4-Oihydrocinnarnic acid were isolated. To investigate the antioxidant activities of each com-pounds, we measured radical scavening activity with DPPH method and anti-lipid peroxidative efficacy on low density lipo-protein (LDL) with TBARS assay and anti-acetylcholinestrase activity with Ellman's Test. Four compounds (ll, IV Vl, Vll) of quercetin glycosides showed significant activity.

효소 저해법을 이용한 Carbamate계 농약의 다성분 잔류분석법 개발 (Development of Multi-Residue Methods for Carbamate Pesticides by the Enzyme Inhibition Test)

  • 김정호
    • 한국환경과학회지
    • /
    • 제17권12호
    • /
    • pp.1325-1330
    • /
    • 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.

Inhibitory effect of Capparis zeylanica Linn. on acetylcholinesterase activity and attenuation of scopolamine-induced amnesia

  • Chaudhary, Amrendra Kumar;Solanki, Ruchi;Singh, Vandana;Singh, Umesh Kumar
    • 셀메드
    • /
    • 제2권2호
    • /
    • pp.19.1-19.6
    • /
    • 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.

Scopolamine 유발 기억 손상 마우스 모델에서 국산 산사와 중국산 산사의 항건망 효과 비교 (Comparative Study on the Effects of Korean and Chinese Crataegus pinnatifida on Scopolamine-induced Memory Impairment in Mice)

  • 이지혜;김혜정;이찬희;박상혁;정철종;백경연;신진기;정지욱
    • 동의생리병리학회지
    • /
    • 제32권6호
    • /
    • pp.375-383
    • /
    • 2018
  • This study was conducted to investigate the cognitive improvement and memory recovery effects of Korean and Chinese Crataegus pinnatifida ethanolic extracts on scopolamine-induced memory impairment in mice. In vivo studies were carried out with mice treated with Korean Crataegus pinnatifida extracts (KCF) and Chinese Crataegus pinnatifida extracts (CCF) in doses of 5 and 50 mg/kg (p.o.) and scopolamine was injected 30 min before the behavioral testing. Antioxidant activity was assessed by 2,2-diphenyl-1-picryl hydrazyl (DPPH) assay and 2,2-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) assay, and acetylcholinesterase inhibition by Ellman's modified method. The chlorogenic acid and hyperoside as marker compounds of KCF and CCF was quantified by ultra-performance liquid chromatography analysis (UPLC). Results showed that KCF was more contained high content of chlorogenic acid and hyperoside than CCF. In addition, KCF was more exerted free radical (DPPH and ABTS) scavenging activity and blocked AChE activity than CCF. In vivo studies also showed that KCF administration has a further improved the memory of scopolamine-treated mice than CCF in Y-maze test, passive avoidance test and Morris water maze test. These results revealed that KCF more prevents scopolamine-induced memory impairments through antioxidant and acethylcholinesterase inhibition effect compared CCF.

Scopolamine 유발 기억 손상 마우스에서 익모초의 효과 (Effect of Leonurus japonicus Houtt. on Scopolamine-induced Memory Impairment in Mice)

  • 이지혜;김혜정;장귀영;서경혜;김미려;최윤희;정지욱
    • 동의생리병리학회지
    • /
    • 제34권2호
    • /
    • pp.81-87
    • /
    • 2020
  • Cognitive impairment is symptoms of dementia, a degenerative brain disease that is drawing attention in a rapidly aging society. This study was conducted to investigate the improvement of cognitive function of Leonurus japonicus on scopolamine-induced memory impairment in mice and the effect and mechanism of memory recovery. In vivo studies were conducted on mice orally pretreated with L. japonicus in doses of 50, 100 and 200 mg/kg (p.o.) and scopolamine (1 mg/kg, i.p.) were injected 30 min before the behavioral task. Antioxidant activity was assessed by 2,2-diphenyl-1-picryl hydrazyl (DPPH) assay and 2,2-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) assay, and acetylcholinesterase (AChE) inhibition activity evaluated by Ellman's method. In behavior studies showed that L. japonicus has an improved the memory of scopolamine-treated mice in Y-maze, passive avoidance and Morris water maze test. In addition, L. japonicus was also exerted free radical scavenging activity and inhibited acetyl cholinesterase activity. These results suggest that L. japonicus improves short-term and long-term memory in scopolamine-induced memory decline model and prevents scopolamine-induced memory impairments through in reduced oxidative stress and acetyl cholinesterase inhibition effect. Thus, L. japonicus is related to functional medicinal materials for prevention and treatment of human dementia patients.

Scopolamine 처리에 의한 인지 및 기억력 손상 마우스에서 박하의 효과 (Mentha arvensis Attenuates Cognitive and Memory Impairment in Scopolamine-treated Mice)

  • 이지혜;김혜정;장귀영;서경혜;김미려;최윤희;정지욱
    • 생약학회지
    • /
    • 제51권1호
    • /
    • pp.70-77
    • /
    • 2020
  • Mentha arvensis is used traditional medicine to treat various disorders. In the present study, M. arvensis were extracted by the solid-phase microextraction (SPME) method and analyzed by gas chromatograph-mass spectrometry (GC-MS). We investigated the protective effects and mechanisms of a M. arvensis extract on scopolamine-induced cognitive and memory impairment. Mice were orally pretreated with a M. arvensis extract or normal saline, and then behavior tests were conducted 30 min after scopolamine injection. The antioxidant capacities were analyzed by free radical scavenging (DPPH and ABTS). Acetylcholinesterase (AChE) activity were also measured using Ellman's method ex vivo test. In behavior tests, percent of spontaneous alteration, escape latency and swimming time in target quadrant were improved by the administration of the M. arvensis extract, which suggests that the M. arvensis extract improves memory function in the scopolamine-treated mice model. In addition, M. arvensis extract showed inhibition of the free radical and AChE activity. The results of the present study suggest that the M. arvensis extract ameliorates scopolamine-induced cognitive and memory deficits through the inhibition of free radicals and AChE activity. Therefore, M. arvensis may be a promising neuroprotective agent for management of learning and memory improvements in human dementia patients.

천연식물자원으로부터 Acetylcholine esterase 저해 활성 탐색 및 인지기능에 미치는 영향 (Screeening of Natural Plant Resources with Acetylcholine esterase inhibitory activity and Effect on Scopolamine-induced Memory Impairment)

  • 최장원;원무호;주한승
    • 농업생명과학연구
    • /
    • 제45권6호
    • /
    • pp.213-226
    • /
    • 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.

Aβ42로 유도된 알츠하이머 마우스 모델에서 이중 가공 인삼열매 추출물의 학습 및 기억 손실 개선 효과 (Double-processed ginseng berry extracts enhance learning and memory in an Aβ42-induced Alzheimer's mouse model)

  • 장수길;안정원;조보람;김현수;김서진;성은아;이도익;박희용;진덕희;주성수
    • 한국식품과학회지
    • /
    • 제51권2호
    • /
    • pp.160-168
    • /
    • 2019
  • AD는 뇌의 신경세포 사멸뿐만 아니라 학습 및 기억능 소실을 초래하는 퇴행성 뇌 질환이며, 기억과 관련된 주요 뇌 구역인 해마(hippocampus)는 콜린성 조절(cholinergic modulation)에 의해 영향을 받는다(Konishi 등, 2015). AD 병인에 대한 다양한 해석과 설명이 있으나 크게는 amyloid cascade hypothesis과 함께 cholinergic hypothesis가 주류를 이루고 있다. 몇몇의 연구에서 ChAT 합성, ACh 분비, nicotin 및 muscarinic 수용체 감소가 AD 뇌의 대뇌피질과 해마에서 관찰되어(Tata 등, 2014) 퇴행성 뇌질환에서의 중요성이 제시되었고, 이를 배경으로 한 acetylcholinesterase inhibitors (AChEI)가 AD 증상 완화의 목적으로 미국 FDA로부터 승인되어 시판되고 있다. 본 연구에서는 인삼의 활성 성분인 진세노사이드가 다량 함유된 PGBC가 $A{\beta}42$로 유도된 치매 모델에서 뇌세포 보호, ACh 분비 증가, 학습력/기억력 증가, ChAT 발현 증가를 확인하여 인삼열매 추출물의 치매 적용 여부를 확인하고자 하였다. 결과에서 언급한 바와 같이 PGBC는 익기 직전의 4년근 인삼 열매에 추출 및 발효 단계를 추가하여 확보된 물질로서, $A{\beta}42$ 섭취, 제거 및 ACh 분비 촉진 활성이 있는 Re, Rd, Rg3 함량이 증가되어(Kim 등, 2014; Jang 등, 2015), PGBC 자체로서 치매 인자에 대한 조절 효능이 예측되었다. 특히, 7증 7포 및 발효과정을 거친 이중가공 인삼 열매 추출물이 비 발효 증포 추출물에 비해 Rg3가 현저히 증가하는 사전 연구결과와 Rg3가 $A{\beta}42$ 제거 활성을 가지는 것으로 확인된 결과를 종합할 때 PGBC 투여가 AD 증상 완화의 조절자 역할을 할 것으로 생각된다(Kim 등, 2013; Jang 등 2015). 본 연구에 따르면, 마우스 치매 모델에 PGBC 처리 시 PAT 및 Morris water-maze test를 통해 대조군 대비 유의한 수준의 인지능력 개선, ACh 합성을 유도하는 ChAT 유전자 발현 증가, ACh 분비량 증가 등이 확인되어 전체적인 인지능 개선에 극적인 영향을 준 것으로 판단된다. 특히 $A{\beta}42$를 뇌 실로 주입(intracranial injection) 하여 나타나는 뇌세포 손상이 PGBC 투여를 통해 보호된 것으로 사료되었으며, 이는 주요 뇌세포 중 하나인 성상세포에서 관찰되는 GFAP 분석을 통해 확인되었다. 뇌 균질액을 이용한 AChE 활성 연구에서도 PGBC가 AChE를 현저하게 저해하는 것으로 확인되어, AD 환자에게 처방이 가능한 2대 의약품 군중 하나인 시냅스 내 신경전달물질 ACh bioavailability 증가 목적의 처방 보조요법 적용이 기대된다(${\check{C}}olovi{\acute{c}}$ 등, 2013). 결론적으로, 본 연구에 사용된 PGBC는 학습 및 기억력을 개선하는 활성물질을 포함하고 있어 1차적인 퇴행성 뇌질환 보조재로서 직간접적인 대증요법 역할과 2차적으로는 뇌 세포 보호를 통한 질병 악화 지연 소재로 개발이 기대되며, 보다 심도 있는 기전연구를 통해 천연물 신소재 개발도 가능할 것으로 사료된다.