• Title/Summary/Keyword: AChE 저해

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

  • Kim, Jung-Ho
    • Journal of Environmental Science International
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    • v.17 no.12
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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.

Effects of Ethanol Extracts from Zingiber officinale Rosc., Curcuma longa L., and Curcuma aromatica Salisb. on Acetylcholinesterase and Antioxidant Activities as well as GABA Contents (생강, 울금, 강황 추출물의 항산화 효과, AChE 억제활성 및 GABA 함량)

  • Jung, Yeon-Seop;Park, Sung-Jin;Park, Jung-Hyun;Jhee, Kwang-Hwan;Lee, In-Seon;Yang, Seun-Ah
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.41 no.10
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    • pp.1395-1401
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    • 2012
  • This study investigated the cognition-related effects on antioxidant activities, ${\gamma}$-aminobutyric acid (GABA) contents, and AChE inhibitory activities in ethanol extracts from Zingiber officinale Rosc. (Korea), Curcuma longa L. (Korea), Curcuma longa L. (Myanmar), and Curcuma aromatica Salisb. (India). These extracts were investigated to determine the relationships among total polyphenols as well as DPPH and ABTS radical scavenging activities, GABA contents, and acetylcholinestrase (AChE) inhibitory activity. For the results, C. longa L. from Myanmar exhibited the highest contents of curcumin (97.1 ${\mu}g/mg$), total polyphenols (98.4 ${\mu}g/mg$), and GABA (1.31 ${\mu}g/mg$), as well as the strongest radical scavenging activities and AChE inhibitory activity. In addition, C. aromatica Salisb from India, which had the highest total polyphenol content (98.4 ${\mu}g/mg$) and strongest radical scavenging activities, exhibited relatively high AChE inhibitory activity similar to that of C. longa L. from Myanmar. On the other hand, Z. officinale Rosc. and C. longa L. from Korea showed low contents of curcumin (12.2 ${\mu}g/mg$) and polyphenols (85.7 ${\mu}g/mg$), as well as low AChE inhibitory activities. However, we could detect 1.11 ${\mu}g/mg$ of GABA in these extracts, which was similar to that of C. longa L. from Myanmar. Therefore, GABA content was not correlated with AChE inhibitory activity. Based on the results, AChE inhibitory activity is highly correlated with polyphenol contents in Zingibearceae family. Overall, among the Zingiberaceae tested, C. longa L. from Myanmar might be a strong candidate as a cognitive-enhancing ingredient.

Screening of Plant Extracts with Cholinesterase Inhibition Activity (콜린 에스테라제 저해효과 보유 식물 추출물 탐색)

  • Park, Saet-Byul;Lee, Jeong-Hoon;Kim, Hyung-Don;Soe, Kyung-Hae;Jeong, Hyeon-Soo;Kim, Dong-Hwi;Lee, Seung-Eun
    • Korean Journal of Plant Resources
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    • v.31 no.5
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    • pp.433-452
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    • 2018
  • This study was conducted to select candidates from plant resources for the purpose of improving or treating Alzheimer's disease, a type of dementia. One hundred and eighty-four plant extracts at a final concentration of $100{\mu}g/ml$ were screened to determine their capacity to inhibit acetylcholinesterase (AChE) by in vitro assay. From this AChE assay, seven plant extracts - including methanol ext. and water ext. of Phellaodendron amurense Rupr. (bark), methanol ext. of Nelumbo nucifera Gaertn (stamen/ovary), methanol ext. of Persicaria tinctoria H. GROSS (flower), methanol ext. of Coptis chinensis (rhizome), ethanol ext. of Cinnamomum cassia Blume(bark) and ethanol ext. of Carthamus tinctorius L. (fruit) - showed effective inhibition activity ranging from 18.7% to 63.1%. The selected extracts were testified their inhibition activities on AChE and BuChE (butyrylcholinesterase) at concentrations of 25, 50, 100, $200{\mu}g/ml$. In the AChE assay, five extracts including methanol ext. of Nelumbo nucifera Gaertn. (stamen/ovary), methanol ext. of Persicaria tinctoria H. GROSS (flower), methanol ext. of Coptis chinensis (rhizome), methanol ext. and water ext. of Phellaodendron amurense Rupr. (bark) showed inhibition activity of 15.0%~73.5%, 19.5%~63.5%, 81.6%~58.5%, 69.9%~80.5%, and 54.8%~78.3%, respectively, at concentrations of 25, 50, 100, $200{\mu}g/ml$. In the BuChE assay, the extracts of Nelumbo nucifera Gaertn. (stamen/ovary), Persicaria tinctoria H. GROSS (flower), and Coptis chinensis (rhizome) showed inhibitory capacities of 58.9~81.6%, 45.8%~72.4%, and 33.1%~55.4% at concentrations of 25, 50, 100, $200{\mu}g/ml$, respectively. In conclusion, it is suggested that Nelumbo nucifera Gaertn. (stamen/ovary), Persicaria tinctoria H. GROSS, Coptis chinensis (rhizome) and Phellaodendron amurense Rupr. (bark) could be selected as candidate materials for improving or treating Alzheimer's disease on the basis of further study.

Antioxidant and Acetylcholinesterase Inhibitory Effect of Aged Raw Garlic Extracts (생마늘 추출 숙성물의 항산화 및 아세틸콜린에스터라아제 저해 효과)

  • Jeong, Hee-Rok;Jeong, Ji-Hee;Jo, Yu-Na;Shin, Jung-Hye;Kang, Min-Jung;Sung, Nak-Ju;Heo, Ho-Jin
    • Journal of agriculture & life science
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    • v.45 no.4
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    • pp.113-120
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    • 2011
  • In vitro antioxidant activities and acetylcholinesterase (AChE) inhibitory effects of solvent fractions from aged raw garlic extracts were investigated. Total phenolics fractioned by hexane, chloroform, ethyl acetate and water from Aged raw garlic extracts were 3.70, 23.63, 31.27 and 2.35 mg/g, respectively. We found that ethyl acetate fractions had the highest in ABTS radical scavenging activities, ferric reducing antioxidant power and inhibitory effect on auto-oxidation of linoleic acid. Intracellular ROS accumulation resulting from $H_2O_2$ treatment of PC12 cells was significantly reduced when ethyl acetate fractions were present in the medium compared to PC12 cells treated with $H_2O_2$ only. In addition, we found that ethyl acetate fractions from aged raw garlic extracts resulted in a dose-dependent manner on AChE inhibition. Consequently, our results suggest that ethyl acetate fractions from aged raw garlic extracts may be useful as decreasing agents of oxidative stress and AChE inhibitors.

A Comparative Study of GABA, Glutamate Contents, Acetylcholinesterase Inhibition and Antiradical Activity of the Methanolic Extracts from 10 Edible Plants (10가지 식용식물 추출물의 가바, 글루탐산 함량, 아세틸콜린에스테라제 억제 및 라디컬 소거능 비교)

  • Jung, Yeon-Seop;Park, Sung-Jin;Kim, Ji-Eun;Yang, Seun-Ah;Park, Jung-Hyun;Kim, Jung-Hyun;Jhee, Kwang-Hwan;Lee, Sam-Pin;Lee, In-Seon
    • Korean Journal of Food Science and Technology
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    • v.44 no.4
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    • pp.447-451
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    • 2012
  • In order to investigate the efficacies for cognitive function of edible plants, we measured the inhibitory effects of acetylcholinesterase(AChE) activity, radical scavenging activities, and the contents of GABA and glutamate in the plant extracts. Among the plant extracts, Schizandra chinensis contained the highest GABA 14.8 mg/g and the extracts of Cnidium officinale and Polygonum multiflorum also had a relatively high GABA. On the other hand, plant extracts except, Acorus gramineus, showed similar glutamate contents. S. chinensis, Hovenia dulcis, Thuja orientalis, and Eleutherococcus senticosus exhibited high inhibition against AChE activities at about 18-33% at 1 mg/mL. In addition, strong radical scavenging activities were also detected in those extracts which showed high AChE inhibition. Taken together, H. dulcis, T. orientalis, E. senticosus, and S. chinensis could be effective resources for enhancing cognitive function. Further, it was suggested that the AChE inhibitory activities of plant extracts might be related to antioxidative activity.

Effect of Organophosphorus Insecticides on the Inhibition of the Acetylcholinesterase Activities (유기인계(有機燐系) 살충제(殺?劑)가 Acetylcholinesterabe 활성저해(活性沮害)에 미치는 영향(影響))

  • Kim, Jung-Ho
    • Applied Biological Chemistry
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    • v.31 no.1
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    • pp.92-99
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    • 1988
  • The responses of brain acetylcholinesterase and plasma cholinesterase activities were examined in chicken given oral doses of an organophosphorus insecticides. Acute oral $LD_{50}$ of terbufos was 1.82mg/kg, and terbufos sulfone was 2.85mg/kg, terbufos sulfoxide, terbufoso xon, terbufosoxon sulfone and terbufosoxon sulfoxide were about $0.30{\sim}0.79mg/kg$, respectively. Acute oral $LD_{50}$ of phorate was 1.02mg/kg, and phorate sulfone was 1.73mg/kg, phorate sulfoxide, phoratoxon, phoratoxon sulfoxide and phoratoxon sulfone were about $0.36{\sim}0.63mg/kg$, respectively. Dosages of 50% inhibition for brain AChE and plasma ChE activities in chicken at 60 mins after oral administration of organop hosphorus insecticides were $35.3{\pm}6.8%\;and\;17.4{\pm}3.6%$ of acute oral $LD_{50}$, respectively. Activities of brain AChE and plasma ChE in chicken at 60 mins after oral administratration of acute oral $LD_{50}$ of organophosphorus insecticides were inhibited by $83.7{\pm}3.9%\;and\;93.3{\pm}2.2%$, respectively.

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Optimal Extraction Condition and Characterization of Antidementia Acetylcholinesterase Inhibitor from Job's Tears (Coix lachrymajobi L.) (율무로부터 항치매성 Acetylcholinesterase 저해물질의 최적추출 조건 및 특성)

  • Seo, Dong-Soo;Jang, Jeong-Hoon;Kim, Na-Mi;Lee, Jong-Soo
    • Korean Journal of Medicinal Crop Science
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    • v.17 no.6
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    • pp.434-438
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    • 2009
  • For the development of a new antidementia functional food or alternative drug using agricultural products, Job's tears (Coix lachrymajobi L.), which shows high acetylcholinesterase (AChE) inhibitory activity (55.1%) was selected and the extraction conditions of AChE inhibitor were optimized. AChE inhibitor of Job's tears was maximally extracted when it was treated with 60% methanol at $40^{\circ}C$ for 6 h. The AChE inhibitor of the methanol extracts was partially purified by systematic solvent extraction, thin layer chromatography, silica gel chromatography and reverse-phase HPLC and the partial purified AChE inhibitor with inhibitory activity ($IC_{50}$) of $0.608\;{\mu}g$ was obtained. The partial purified AChE inhibitor was soluble in methanol and hexane, and insoluble in water. Its maximum absorption spectra was 230 nm and also it was stable in the range of $30^{\circ}C$ and $70^{\circ}C$ and pH 4.0-8.0 for 1 h.

Acetylcholinesterase Inhibitory and Antioxidant Properties of Aster yomena Extract (쑥부쟁이 추출물의 아세틸콜린에스테라제 저해 및 항산화 활성)

  • Bae, Jong-Sup;Kim, Tae-Hoon
    • The Korea Journal of Herbology
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    • v.24 no.4
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    • pp.121-126
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    • 2009
  • Objectives : To evaluate the radical scavenging and acetylcholinesterase (AChE) inhibitory activities of the ethylacetate (EtOAc)-soluble portion of a methanolic extract of Aster yomena, three different assay systems were performed. Methods : The antioxidant activity of A. yomena extract was tested as its capacity to scavenging free radicals of DPPH and $ABTS^+$, which has been widely used to evaluate the antioxidant activity of natural products from plant sources. AChE inhibitory activity was tested against mouse brain AChE by spectrophotometric method of Ellman using ELISA microplate reader. Results : The methanolic extract of A. yomena was fractionated and the EtOAc-soluble portion showed significant AChE inhibitory and free radical scavenging effects. Also the EtOAc-soluble portion revealed the highest phenolic contents as compared to the other extracts. Conclusions : These results indicate that phenolic compounds may be important constituents that give rise to the anti-AChE and antioxidative activities of A. yomena extract. Further phytochemical studies on this plant, for nutraceutical or pharmaceutical application, are warranted.

Extraction of Acetylcholinestrase from the Housefly and Three Other Insect Species for In Vitro Anticholinesterase Screening (In Vitro Anticholinesterase 스크리닝을 위한 집파리 및 3종 곤충으로부터의 Acetylcholinesterase의 추출)

  • 이시혁;이준호;조광연
    • Korean journal of applied entomology
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    • v.30 no.1
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    • pp.18-28
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    • 1991
  • The optimal pH of the extraction buffer was 7.5 considiering AChE stability and its buffer capacity when AChE was isolated and extracted from the housefly(Musca domesitca L.)and three other insect species with 0.01 M sodium phosphate buffer. Also, the optimal pH of the reaction buffer was 7.5 considering enzyme activity and its buffer capacity when AChE activity was measured with the substrate in 0.1 M sodium phosphate buffer. The Potter Elvehjem type homogenizer with Teflon pestle was used to homgenize the tissues. When preparing a AChE suspension by centrifuging the homogenate, 700 g supernatant of adult head for the housefly, 700 g supernatnat of 5th instar nymphal whole body for the brown planthopper, lipid-eliminated 10,000 g supernatant of 5th instar larval whole body for the diamondback moth, and 700 g supernatant of 4th instar larval head for the tobacco cutworm were considered satisfactory as enzyme sources in view of mass preparation, extraction efficiency and stability of enzyme activity during evaluation. When AChE suspensions of 4 insect species were stored at $-18^{\circ}C$, more than 90% of activity was maintained up to 3 weeks. Km values of AChEs of the housefly, the brown planthopper, and the diamondback moth were 0.042, 0.037 and 0.043 mM, respectively and AChE-specific substrate inhibition was observed at high concentration. Km value of the tobacco cutworm ChE was 1.15 mM and BuChE characteristics was observed, though further study is needed. The optimal substrate concentration for the AChE inhibition tests was 0.5 mM for the housfly, the brown planthopper, and the diamondback moth and 12 mM for the tobacco cutworm.

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Inhibitory Effect of Bee Venom Herbal Acupuncture Solution on Acetylcholinesterase in PC 12 Cells (PC12 세포에서 봉독약침액의 Acetylcholinesterase 억제효과)

  • Choi, Yang-Sik;Kim, Jong-In;Choi, Do-Young;Lee, Jae-Dong;Koh, Hyung-Kyun
    • Journal of Acupuncture Research
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    • v.25 no.2
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    • pp.1-9
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    • 2008
  • 목적 : Acetylcholine은 콜린과 아세트산의 에스테르로 인체에서 중요한 신경전달물질로 Acetylcholine-sterase(AChE)라는 효소에 의해 분해된다. Alzheimer's disease, ataxia, myasthenia gravis, Parkinson' disease 등의 질환에 AChE 억제제가 사용되어 왔으며 최근 한약재의 AChE 억제 효능에 관한 연구들도 진행되고 있다. 봉독은 관절염, 통풍 등의 질환에 응용되어 왔으며 진통효과 및 항염증작용에 대한 임상적, 실험적 연구가 많이 보고되어 왔으나 AChE 억제효과에 대한 연구는 아직까지 보고된 바 없다, 본 연구에서는 봉독약침액과 봉독의 과민반응 유발항원 중 하나인 Phospholipase A2 억제효능이 있는 것으로 알려진 상백피를 혼합한 상백피봉독약침액의 AChE 억제효과를 알아보았다. 방법 : PC12 세포주에서 추출한 AChE와 0.1, 0.01 and $0.001mg/m{\ell}$ 농도의 봉독약침액 및 상백피봉독약침액을 60분간 반응시켰다. 효소면역측정법(ELISA)을 이용하여 흡광도를 10분, 30분, 60분 경과시 각각 측정한 후 효소활성저해도(%)를 계산하였다. 효소활성저해도(%) = [(Cc - Ce)/Cc] ${\times}$ 100 Cc : 대조군 흡광도, Ce : 실험군 흡광도 결과 : 1. 봉독약침액은 0.1, 0.01, $0.001m{\ell}/mg$의 농도에서 30분 경과 후부터 유의성 있는 억제효과를 나타내었다. 2. 상백피봉독약침액은 $0.1m{\ell}/mg$ 농도에서 10분 경과 후부터 유의성 있는 억제효과를 나타내었고, $0.01m{\ell}/mg$ 농도에서 30분경과 시 유의성 있는 억제효과를 나타내었다. 3. 봉독약침액과 상백피봉독약침액의 AChE 억제효과 비교에서 봉독약침액의 억제효과가 상백피봉독약침액 보다 뛰어났다. 요약 : 봉독약침액과 상백피봉독약침액의 AChE 억제효과를 확인하여 두 군 모두 유의성 있는 결과를 얻을 수 있었다. 앞으로 알츠하이머병이나 치매와 같은 신경퇴행성 질환에 대한 봉독의 임상적 활용 및 보다 넓은 범위에 대한 연구가 필요할 것이라 사료된다.

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