현호색의 Acetylcholinesterase 활성 저해 성분 및 그 작용기전

황세영,장영표,변순정,전미희,김영중,Hwang, Se-Young,Chang, Young-Pyo,Byun, Soon-Jung,Jeon, Mee-Hee,Kim, Young-Choong 한국생약학회 1996 생약학회지 Vol.27 No.2

In the course of searching for acetylcholinesterase inhibitors from crude drugs, it was found that total MeOH extract of Corydalis Tuber showed significant inhibitory effect on acetylcholinesterase. To isolate acetylcholinesterase inhibitors from Corydalis Tuber, total MeOH extract of the the crude drug was subjected to activity guided fractionation. The MeOH extract was suspended in water and fractionated with methylene chloride and subjected to acid-base fractionation. Silica gel column chromatography of the basic fraction which showed significant inhibitory effect on acetylcholinesterase was carried out and 5 subfractions (1-5) were obtained. From subtraction 4, compound I was isolated. The structure of isolated compound I was identified by spectroscopic parameters of $^1H-NMR$, $^{13}C-NMR$, EI-MS and FAB-MS. The compound I was identified as berberine. It was found from the Lineweaver-Burk plot that berberine was a reversible and specific inhibitor of acetylcholinesterase having 90% inhibitory effect at the concentration of $2.5{\mu}M$.

운동과 블루베리 섭취가 알츠하이머 치매에 미치는 영향분석

손원목,곽이섭 한국스포츠학회 2016 한국스포츠학회지 Vol.14 No.4

고령화 사회로 접어들면서 치매 유병률이 빠르게 증가 하고 있으며, 이중 알츠하이머 치매는 점진적인 기억, 판단 및 언어능력 등 지적인 기능이 감퇴된다. 알츠하이머 치매의 발병 원인에 관한 명확한 기전이 밝혀지지 않았으나, Amyloid-β의 응집, 신경원섬유덩굴의 형성, acetylcholinesterase의 활성으로 acetylcholine 농도 저하 등이 원인 이 되고 있다. 블루베리는 탄수화물, 무기질 등을 함유하고 있으며, 특히 안토시아닌, 플라보노이드 등이 풍부하고 그 중 안토시아닌은 주목할 만하다. 안토시아닌은 뇌의 신경체계를 활성화하여 기억기능을 증가와 해마에서 안토시아닌의 분 포는 신경 신호 증가와 관련을 보고 하였다. 블루베리를 섭취한 결과 Amyloid-β 감소, acetylcholinesterase의 활성 억제 및 산화스트레스 감소를 보고 하였다. 운동은 노화로 인한 신경세포생성의 감소율을 억제시키며, 노화로 인한 인지 기능의 손상을 약화 시키고, 운동 후 Amyloid-β 감소, acetylcholinesterase의 활성 억제가 보고되었다. 따라서, 고 령화로 인해 증가 되는 치매환자의 예방 및 치료를 위하여 다양한 천연식물과 치매노인을 위한 운동프로그램에 관한 연 구가 이루어 져야 할 것으로 사료된다. As the society is aging, the prevalence of dementia is rapidly increasing. Among the types of dementia, Alzheimer’s disease brings gradual deterioration of intellectual functions such as memory, judgment, and the faculty of speech. Although the specific mechanism of the pathogenesis of Alzheimer’s disease is not known, aggregation of Amyloid-β, decrease in the concentration of acetylcholine due to the activation of acetylcholinesterase, and activation of free radicals are cited as the possible causes. Blueberries contain carbohydrates and minerals, and are particularly rich in anthocyanins and other flavonoids; here anthocyanins are especially noteworthy. They activate the nerve system of the brain and improve memory function. After consumption of blueberries, reduction in Amyloid-β, inhibition of acetylcholinesterase activation, and decrease in oxidative stress could be observed. Exercise can lead to a decrease in aging-related processes, like neuronal and cognitive impairment. Both reduction of Amyloid-β and inhibition of acetylcholinesterase activation have been reported after exercising. Therefore, to prevent the onset of dementia and treat patients already affected studies on various natural plants and specific exercise programs for elderly patients with dementia are thought to be necessary.

Modification of Substrate Inhibition of Synaptosomal Acetylcholinesterase by Cardiotoxins

( Seyed Omid Ranaei Siadat ),( Gholam Hosein Riazi ),( Mehdi Sadeghi ),( Long Sen Chang ),( Shinne Ren Lin ),( Peyman Eghtesadi Araghi ),( Gholam Hossein Hakimelahi ),( Ali Akbar Moosavi Movahedi ) 생화학분자생물학회 2004 BMB Reports Vol.37 No.3

Different types of cardiotoxin (I-V and n) were isolated and purified from the venom of the Taiwan cobra (Naja naja atra). The effects of these cardiotoxins were studied on membrane-bound acetylcholinesterase, which was isolated from a sheep`s brain cortex. The results showed that cardiotoxins I-III, V, and n activated the enzyme by modification of substrate inhibition, but cardiotoxin IV`s reaction was different. The inhibition and activation of acetylcholinesterase were linked to the functions of the hydrophobicity index, presence of a cationic cluster, and the accessible arginine residue. Our results indicate that Cardiotoxins have neither a cationic cluster nor an argininge residue in their surface area of loop I; therefore, in contrast to fasciculin, cardiotoxins are attached by loop II to the peripheral site of the enzyme. As a result, fasciculin seems to stabilize nonfunctional conformation, but cardiotoxins seem to stabilize the functional conformation of the enzyme. Based on our experimental and theoretical findings, similar secondary and tertiary structures of cardiotoxins and fasciculin seem to have an opposite function once they interact with acetylcholinesterase. s

지렁이에서 추출한 Acetylcholinesterase, Cytochrome P₄₅₀, and Heat Shock protein 70을 이용한 유기성슬러지 독성 평가

나영은,방혜선,김명현,김민경,노기안,이정택,안용준,윤성탁,Na, Young-Eun,Bang, Hae-Son,Kim, Myung-Hyun,Kim, Min-Kyoung,Roh, Kee-An,Lee, Jung-Taek,Ahn, Young-Joon,Yoon, Seong-Tak Korean Society of Soil Science and Fertilizer 2007 한국토양비료학회지 Vol.40 No.4

4 종류의 폐기물(생활하수오니, 공단하수오니, 피혁오니, 주정오니)과 대조구로서 돈분퇴비가 지렁이에게 미치는 독성을 평가하기 위하여 대표적인 유해성 평가 biomarker 3종류 (acetylcholinesterase, cytochrome $P_{450}$, heat shock protein 70)를 사용하였다. 유기성 폐기물에 대한 acetylcholinesterase의 활성은 돈분퇴비의 경우 활성이 약간 촉진된 반면 생활하수오니, 공단하수오니, 피혁오니, 주정오니는 영향을 미치지 않았다. Cytochrome $P_{450}$의 활성은 공단하수오니와 피혁오니는 활성을 억제하였고 생활하수오니, 주정오니, 돈분퇴비는 영향을 미치지 않았다. 또한 Hsp70의 발현량은 증류수보다 돈분퇴비는 1.9배, 주정오니는 3.0배, 생활하수오니는 3.3배, 공단오니는 4.4배, 피혁오니는 4.7배 순으로 지렁이 (Eisenia fetida)에게 스트레스를 많이 주었다. 이상의 결과로부터, 4 종류의 폐기물(생활하수오니, 공단하수오니, 피혁오니, 주정오니)은 돈분퇴비보다 독성이 강한 것으로 판단하였다. 또한 AChE, Cytochrome $P_{450}$과 Hsp70은 추후 유기성 폐기물의 유해성을 모니터링하기에 적합한 biomarker로서 가치가 있다고 생각한다. The toxicitiy of organic sludge such as municipal sewage sludge (MSS), industrial sewage sludge (ISS), alcohol fermentation processing sludge (AFPS) and leather processing sludge (LPS) were evaluated with three environmental biomarkers as acetylcholinesterase, cytochrome P450, and heat shock protein 70 extracted from earthworm (Eisenia fetida). Their toxicities were compared with those of pig manure compost (PMC). MSS, ISS, LPS, and AFPS did not significantly affect the acetylcolinesterase activity, whereas only the elutriate of PMC slightly was increased the activity. MSS, AFPS, and PMC tended to slightly inhibit the cytochrome $P_{450}$ activity, but ISS and LPS showed significantly the inhibitory effect on cytochrome $P_{450}$. The hsp70 expression began to increase after treatments and showed high induction at 6 hour, followed by zero level at around 12 hour. The quantity of the hsp70 expressed by elutriate treatments of PMC, AFPS, MSS, ISS, and LPS was 1.9, 3.0, 3.3, 4.4, and 4.7 fold higher than that of distilled water. These results indicate that in toxicity tests of five organic waste materials, four kinds of sludge materials appeared more toxic than PMC. Results of AChE, P450, and hsp70 of earthworm might be useful for expecting or assessing an effect by exposure of organic wastes to earthworms in soil.

Cholinesterases inhibition studies of biological active compounds from the rhizomes of <i>Alpinia officinarum</i> Hance and <i>in silico</i> molecular dynamics

Lee, Ji Sun,Kim, Jang Hoon,Han, Yoo Kyong,Ma, Jin Yeul,Kim, Young Ho,Li, Wei,Yang, Seo Young Elsevier 2018 INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES Vol.120 No.2

<P><B>Abstract</B></P> <P>Six diarylheptanoids (<B>1</B>–<B>6</B>) and two flavonoids (<B>7</B> and <B>8</B>) derived from <I>Alpinia officinarum</I> were evaluated for their ability to inhibit acetylcholinesterase. Compound <B>1</B> showed the highest degree of inhibition, with an IC<SUB>50</SUB> of approximately 2 μM, followed by moderate degrees of inhibition by <B>2</B>, <B>4</B> and <B>7</B>, with IC<SUB>50</SUB> values ranging from 20 to 40 μM. The remaining isolated compounds <B>3</B>, <B>5</B>, <B>6</B> and <B>8</B> had IC<SUB>50</SUB> values greater than 50 μM. Enzyme kinetic studies showed that the compounds with high or moderate activity were competitive inhibitors, anchored to the active site of acetylcholinesterase. In particular, compounds <B>1</B> and <B>2</B> were docked at slightly different positions from those occupied by <B>4</B> and <B>7</B>. Furthermore, molecular dynamics studies showed that compound <B>1</B> maintained its interactions with residues Thr74 and Phe295 throughout the simulation trajectory. Our findings suggest that compound <B>1</B> is a potential therapeutically relevant inhibitor of acetylcholinesterase.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Six diarylheptanoids (<B>1</B>–<B>6</B>) and two flavonoids (<B>7</B> and <B>8</B>) were isolated from <I>Alpinia officinarum</I>. </LI> <LI> Compound <B>1</B> showed acetylcholinesterase inhibitory activity with IC<SUB>50</SUB> value of approximately 2 μM. </LI> <LI> Compounds <B>1</B>, <B>2</B>, <B>4</B>, and <B>7</B> inhibited the catalytic reaction of acetylcholinesterase as competitive mode. </LI> <LI> Computational simulation study suggested the predicted binding position of the compound <B>1</B> with catalytic site of receptor. </LI> </UL> </P>

조희풀(Clematis heracleifolia) 지하부의 성분과 Acetylcholinesterase억제 활성

김미애,김명조,전완주,권용수 한국생약학회 2015 생약학회지 Vol.46 No.1

To clarify chemical constituents of Clematis heracleifolia, isolation and structure elucidation of the underground parts of C. heracleifolia were performed. Five compounds were isolated from CHCl3 and n-BuOH soluble fraction of this plant. On the basis of spectral and physico-chemical data, the structure of isolated compounds were identified as coniferyl alcohol (1), scoparone (2), (+)-lariciresinol (3), phytosterols (4), and daucosterol (5), respectively. All compounds are isolated from this plant for the first time. To evaluate anti-acetylcholinesterase activity of the isolated compounds, compounds 1, 2, 3, and 5 were tested inhibition activity against acetylcholinesterase. Among tested compounds, daucosterol (5) showed acetylcholinesterase inhibitory activity with IC50 value of 6.1 mM.

증례 : 양수검사에서 AFP, AchE가 양성이였으나 정밀 초음파 추적검사 후 정상아를 분만한 1예

박혜리 ( Hye Ri Park ),전혜선 ( Hye Sun Jun ),차동현 ( Dong Hyun Cha ),박상원 ( Sang Won Park ),정창조 ( Chang Jo Chung ) 대한주산의학회 2006 大韓周産醫學會雜誌 Vol.17 No.4

신경관 결손증은 전세계적으로 약 1,000명당 1.4~2명의 발생빈도를 보이는 중요한 선천성 기형 중 하나로 알려져 있다. 선별검사(screening test)로서 모체혈청 AFP이 일반적으로 이용되고 있다. 그러나 이는 다태 임신, 태아 사망, 잘못 계산 된 임신주수, 낭성 임파관종, 기형종, 신장 이상, 식도 폐쇄증, 태반 이상 등 여러가지 경우에서 나타날 수 있으므로 일단 모체 혈청의 AFP이 증가되었다면 일단 임신주수, 태아의 생존여부, 태아의 수 등을 확신하기 위해서 우선 정밀 초음파 검사를 해야 하고, 이러한 경우를 제외한 후에 신경관 결손증이 의심되거나 확실하지 않은 경우 양수 내 AFP와 AchE를 측정한다. 그러나 양수 내 AFP은 위양성율이 높고, 이를 보완하기 위한 양수 내 AchE도 0.3%정도로 위양성율이 존재하므로 이때 정밀초음파에서 이상 소견 보이지 않을 때 과거력이나 유전력 등의 세밀한 상담 후 정기적인 정밀초음파 검사로 추적검사를 시행하면서 임신을 유지할 수 있다. 이에 본 저자는 임신 17주에 시행한 모체 혈청 AFP이 양성이 나와 시행한 양수 AFP, AchE이 양성이 나왔으나 시행한 정밀 초음파상 신경관, 복벽, 기타장기 내 기형 소견 보이지 않아, 가족력이나 과거력, 세밀한 유전력 상담 후 정기적인 정밀 초음파로 추적 검사하여 정상아를 분만한 한 1예를 보고 하는 바이다. Neural tube defects are reported one of the important congenital malformation in the world, with an incidence of 1.4 to 2 per 1000 pregnancies. Maternal serum AFP at the second trimester is used as screening test. But this is associated with numerous causes, including twins, fetal death, misdated pregnancies, cystic hygromas, teratomas, renal abnormalities, esophageal atresia and aberrations in the placenta. If the AFP level was elevated, targeted ultrasonography should be evaluated for confirming the gestational age and fetal viability, fetal number. Then if the ultrasonographic examination is nondiagnostic, or if an NTD is suspected, amniotic fluid AFP should be measured with acetylcholinesterase. But the false positive rate of the AF-AFP is high, and there is 0.3% of the false positive rate in amniotic acetylcholinesterase. This time the women with normal ultrasonographic examination continue the pregnancy after counseling of family history, past history with follow up of ultrasonography. We report a case with elevated second trimester MS-AFP, AF-AFP and positive amniotic acetylcholinesterase, but in which repeated sonographic findings were normal and result in delivery of a healthy baby without anomalies.

Hydrophobic Interaction Between the Acyl Moiety of Choline Esters and the Active Site of Acetylcholinesterase

Myung, Pyung-Keun,Sok, Dai-Eun 충남대학교 약학대학 의약품개발연구소 1995 藥學論文集 Vol.11 No.-

Existence of a binding site for choline esters with an acyl chain of various sizes was examined by comparing the inhibitory potency of the respective compound. In contrast to acetylcholine, which showed a pure competitive pattern of inhibition, choline esters with an acyl chain of a long size (C≥5) expressed a mixed type of inhibition. Binding of choline esters containing a long chain (C_7-C_12) to the hydrophobic region in the active site is deduced from a linear relationship between the K_iE value and the size of acyl moiety, and a good hydrophobicity relationship. In addition, the non-competitive component in the inhibition of acetylcholinesterase seems to be due to the interaction of choline esters with both the hydrophobic site and the trimethylammonium-binding site in the active center of the acetylated acetylcholinesterase.

Vincristine이 Mouse 장관내 근층간 신경총의 Acetylcholinesterase 활성에 미치는 영향

이규식 한양대학교 의과대학 1991 한양의대 학술지 Vol.11 No.2

Vincristine, one of vinca alkaloids, is widely used for chemotherapy of a variety of malignancy. The mechanism of anticancerous activity of this agent is based of ingivition of mitoses by interference of development of microtobule of systheses of nucleic acid and protein and of destruction of sytoplasic membrane and microtubule. However, this inhibirtory ettect of cancer call proliferation is non-specific, so clinical use of vincristine is frequently limited due to its effect on the hematopoietic and nervous system especially. The author had undertaken this study ot pursue the effect of vincristine on the parasympathetic ganglia in intestines histochemically. Totals of 20 healthy make ICR mouse weighing about 20gm were used as experimentla animals. The experimental animals were divided into normal control and vincristine treated groups. Mice of vincristine treated group were administered intraperitoneally 25mg of vincristine/kg of body weight of mouse diluted in 0.2ml of distilled water once per a day during 5 days. Mice of control group were treated with normal saline as same dose and method as above. 24hours after the last administration, the animals were sacrificed by carotid exangination and then mid-protions of jejunum and transverse colon was obtained respectively in 1cm lenhth. As soon as the specimens were collected, they were transported into cryostat and frozen at -20℃. Frozen sectioned preparations 16㎛ in thicknees were treated by Gerebtzoff method (1959) for acetylcholinesterase. All preparations were observed with light microscope. The results were as floolws. 1.Acetylcholinesterase activity of submucosal and myenteric plexuses in jejunum of normal control mice were observed as moderately positive, but that of vincristine treated mice were weakly positive. 2.Acetylcholinesterase activity of submucosal and myenteric plexuses in transverse colon of normal control mice were abserved as strongly positive but that of vincristine treated mice were tracely positive. It is consequently suggested that vincristine would damage on the Myenteri plexus in mice.

Hydrophobic Interaction Between the Acyl Moiety of Choline Esters and the Active Site of Acetylcholinesterase

Myung, Pyung-Keun,Sok, Dal-Eun 충남대학교 생물공학연구소 1996 생물공학연구지 Vol.4 No.-

Existence of a binding site for choline esters with an acyl chain of various sizes was examined by comparing the inhibitory potency of the respective compound. In contrast to acetylcholine, which showed a pure competitive pattern of inhibition, choline esters with an acyl chain of a long size (C≥5) expressed a mixed type of inhibition. Binding of choline esters containing a long chain (C_7-C-12) to the hydrophobic region in the active site is deduced from a linear relationship between the K_1E value and the size of acyl moiety, and a good hydrophobicity relationship. In addition, the non-competitive component in the inhibition of acetylcholinesterase seems to be due to the interaction of choline esters with both the hydrophobic site and the trimethylammonium-binding site in the active center of the acetylated acetylcholinesterase.