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acetylcholinesterase Biosensor Based on Poly(diallyldimethylammonium chloride)-multi-walled Carbon Nanotubes-graphene Hybrid Film

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Nano-Micro Letters 2013年第1期5卷 47-56页

作者： Xia Sun Zhili Gong Yaoyao Cao Xiangyou Wang College of Agriculture and Food Engineering Shandong University of Technology

In this paper, an amperometric acetylcholinesterase(ACh E) biosensor for quantitative determination of carbaryl was developed. Firstly, the poly(diallyldimethy-lammonium chloride)-multi-walled carbon nanotubes-graphene hybrid film was modified onto the glassy carbon electrode(GCE) surface, then ACh E was immobilized onto the modified GCE to fabricate the ACh E biosensor. The morphologies and electrochemistry properties of the prepared ACh E biosensor were investigated by using scanning electron microscopy, cyclic voltammetry and electrochemical impedance spectroscopy. All variables involved in the preparation process and analytical performance of the biosensor were optimized. Based on the inhibition of pesticides on the ACh E activity, using carbaryl as model compounds, the biosensor exhibited low detection limit, good reproducibility and high stability in a wide range. Moreover, the biosensor can also be used for direct analysis of practical samples, which would provide a new promising tool for pesticide residues analysis.

关键词： Biosensor acetylcholinesterase Multi-walled carbon nanotubes Graphene Poly(diallyldimethy lammonium chloride)

acetylcholinesterase inhibition ameliorates retinal neovascularization and glial activation in oxygen-induced retinopathy

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International Journal of Ophthalmology(English edition) 2020年第9期13卷 1361-1367页

作者： Qiu-Ping Liu Xian Zhang Ya-Zhou Qin Jing-Lin Yi Jing-Ming Li Affiliated Eye Hospital of Nanchang University Nanchang 330006Jiangxi ProvinceChina Department of Ophthalmology First Affiliated Hospital of Xi'an Jiaotong UniversityXi’an 710061Shaanxi ProvinceChina

AIM:To investigate whether inhibition of acetylcholinesterase(AChE)by donepezil ameliorate aberrant retinal neovascularization(RNV)and abnormal glial activation in oxygen-induced retinopathy(OIR).METHODS:A mouse model of RNV was induced in postnatal day 7(P7)mice by exposure to 75%oxygen.Donepezil was administrated to P12 mice by intraperitoneal injection.Expression and localization of AChE in mouse retinas were determined by immunofluorescence.RNV was evaluated by paraffin sectioning and hematoxylin and eosin(HE)staining.Activation of retinal M uller glial cells were examined by immunoblot of glial fibrillary acidic protein(GFAP).rMC-1.a retinal Muller cell line,was used for in vitro study.Expression of hypoxia-induced factor 1α(HIF-1α)and vascular endothelial growth factor(VEGF)were determined by Western-blot analysis,enzyme-linked immunosorbent assay(ELISA)or immunostaining.RESULTS:Aberrant RNV and glial activation was observed after OIR.Of note,retinal AChE was mainly expressed by retinal Muller glial cells and markedly increased in OIR mice.Systemic administration of donepezil significantly reduced RNV and abnormal glial activation in mice with OIR.Moreover,ischemia-induced HIF-1αaccumulation and VEGF upregulation in OIR mouse retinas and cultured rMC-1 were significantly inhibited by donepezil intervention.CONCLUSION:AchE is implicated in RNV with OIR.Inhibition of AChE by donepeizl is likely to be a potential therapeutic approach for retinal neovascular diseases.

关键词： acetylcholinesterase oxygen-induced retinopathy neovascularization glosis mice

Highly selective NIR fluorescent probe for acetylcholinesterase and its application in pesticide residues detection

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Chinese Chemical Letters 2022年第9期33卷 4233-4237页

作者： Shengui He Shufang Zhang Xin Zhao Xinyue Zhu Lisen Chen Jingnan Cui State Key Laboratory of Fine Chemicals Dalian University of Technology Dalian 116024China

A series of near-infrared(NIR)fluorescent substrates(NDRO-1∼8)derived from fluorophore NDRH with different volumes of ester bond as the recognition group were designed and synthesized for the detection of acetylcholinesterase(AChE),among which NDRO-1 with the smallest acetate group displayed the highest activity toward AChE.The detection limit of NDRO-1 for sensing AChE was 0.32μg/mL,and K_(m )was 6.40μmol/L,indicating ultra-sensitivity and good affinity of NDRO-1 toward AChE.NDRO-1 was used to detect the inhibitory of four kinds of pesticides including methamidophos,dichlorvos,and the detection limit was lower than 50μg/L,which was further used in pesticide residues detection.

关键词： NIR fluorescent probe acetylcholinesterase Enzyme-activated Pesticide residue detection

Novel insights on acetylcholinesterase inhibition by Convolvulus pluricaulis,scopolamine and their combination in zebrafish

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Natural Products and Bioprospecting 2022年第1期12卷 56-70页

作者： Kalyani Bindu Karunakaran Anand Thiyagaraj Kirankumar Santhakumar Supercomputer Education and Research Centre Indian Institute of Sci-enceBengaluruIndia Department of Genetic Engineering SRM Institute of Science and TechnologyKattankulathur 603203India Zebrafish Genetics Laboratory Department of Genetic EngineeringSRM Institute of Science and TechnologyKattankulathur 603203India

acetylcholinesterase(AChE)inhibitors increase the retention of acetylcholine(ACh)in synapses.Although they allevi-ate cognitive deficits in Alzheimer’s disease,their limited benefits warrant investigations of plant extracts with similar properties.We studied the anti-AChE activity of Convolvulus pluricaulis(CP)in a zebrafish model of cognitive impair-ment induced by scopolamine(SCOP).CP is a perennial herb with anti-amnesiac and anxiolytic properties.It contains alkaloid,anthocyanin,coumarin,flavonoid,phytosterol and triterpenoid components.Isoxazole(ISOX)was used as a positive control for AChE inhibition.CP-treated 168 hpf larvae showed a similar pattern of AChE inhibition(in the myelencephalon and somites)as that of ISOX-treated larvae.CP was superior to ISOX as evidenced by the retention of avoidance response behavior in adult zebrafish.Molecular docking studies indicated that ISOX binds Ser203 of the catalytic triad on the human AChE.The active components of CP-scopoletin and kaempferol-were bound by His447 of the catalytic triad,the anionic subsite of the catalytic center,and the peripheral anionic site.This suggested the ability of CP to mediate both competitive and non-competitive modes of inhibition.Surprisingly,SCOP showed AChE inhibition in larvae,possibly mediated via the choline-binding sites.CP+SCOP induced a concentration-dependent increase in AChE inhibition and ACh depletion.Abnormal motor responses were observed with ISOX,CP,ISOX+SCOP,and CP+SCOP,indicative of undesirable effects on the peripheral cholinergic system.Our study proposes the examination of CP,SCOP,and CP+SCOP as potential AChE inhibitors for their ability to modulate cognitive deficits.

关键词： Alzheimer’s disease acetylcholinesterase Zebrafish Convolvulus pluricaulis Scopolamine Isoxazole

Enhanced oxidase-like activity of g-C_(3)N_(4) nanosheets supported Pd nanosheets for ratiometric fluorescence detection of acetylcholinesterase activity and its inhibitor

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Chinese Chemical Letters 2022年第2期33卷 757-761页

作者： Chenghui Zhang Pengjuan Ni Bo Wang Wendong Liu Yuanyuan Jiang Chuanxia Chen Jian Sun Yizhong Lu School of Materials Science and Engineering University of JinanJi’nan 250022China State Key Laboratory of Electroanalytical Chemistry Changchun Institute of Applied ChemistryChinese Academy of SciencesChangchun 130022China

The undesirable enzymatic activity of nanozymes under near neutral p H condition and the traditional single signal output always restrict the analytical application of nanozyme-based biosensors.Herein,graphitic carbon nitride nanosheets supported palladium nanosheets composite (Pd/g-C_(3)N_(4)) with both oxidase-like activity and fluorescent property is synthesized.Notably,Pd/g-C_(3)N_(4)exhibits enhanced oxidase-like activity compared to Pd NSs under p H 7.4.By combining Pd/g-C_(3)N_(4)with o-phenylenediamine(OPD),a ratiometric fluorescence assay for acetylcholinesterase (ACh E) activity detection is developed.Pd/g-C_(3)N_(4)can catalyze oxidation of nonfluorescent OPD to fluorescent oxidized OPD (ox OPD,Em=565nm),which can quench fluorescence of g-C_(3)N_(4)supporter (Em=441 nm) through fluorescence resonance energy transfer (FRET).However,in presence of ACh E,acetylthiocholine can be hydrolyzed into thiocholine,which will block the oxidase-like activity of Pd/g-C_(3)N_(4)and then hamper the FRET process.This ratiometric fluorescence assay is also viable to screen ACh E inhibitor.This work will guide design of ratiometric fluorescence assay based on nanozymes with improved enzymatic activity.

关键词： Oxidase-like activity Ratiometric fluorescence acetylcholinesterase g-C_(3)N_(4)nanosheets Pd nanosheets

Efficacy of Silene arenosa extract on acetylcholinesterase in Bungarus sindanus(krait) venom

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Journal of Traditional Chinese Medicine 2021年第3期41卷 349-354页

作者： Adil Shah Mushtaq Ahmed Naila Sher Nadia Mushtaq Rahmat Ali Khan Fozia Midrarullah Department of Biotechnology Faculty of Biological SciencesUniversity of Science&TechnologyBannu 28100KPK-Pakistan Department of Botany Faculty of Biological SciencesUniversity of Science&TechnologyBannu 28100KPK-Pakistan Department of Biochemistry KMU Institute of Medical SciencesKohat 26000Pakistan Department of Biotechnology Saheed Benazir Bhutto UniversitySheringalDir UpperKPK-Pakistan

OBJECTIVE:To examine the efficacy of Silene arenosa extract on acetylcholinesterase (AChE) of krait (Bungarus Sindanus) snake venom.METHODS:The present project designed to evaluate the inhibition of AChE by following standard procedures.RESULTS:Statistical analysis of the results showed that Silene arenosa exerted 73%inhibition against the krait venom acetylcholinesterase at fixed substrate acetylcholine (ACh) concentration (0.5 mM).Kinetic analysis using the Lineweaver Burk plot revealed that Silene arenosa caused a competitive type of inhibition i.e.K_(m) values increased from 26.6 to 93.3 mM (26.6%to 93.3%) and V_(max)remained constant in a concentration-dependent manner.Silene arenosa competes with the substrate to bind at the active site of the enzyme.The K_(mapp)of venom AChE for Silene arenosa increased from 60%to 81.6%and the V_(maxapp)remains constant.K_(i)(inhibition constant was estimated to be 48μg for snake venom);while the K_(m)(Michaelis-Menten constant of AChE-substrate into AChE and product) was estimated to be 0.5 mM.The IC_(50)of AchE calculated for Silene arenosa was 67μg.CONCLUSION:The present results suggest that Silene arenosa extract can be considered as an inhibitor of snake venom AChE.

关键词： acetylcholinesterase snake venoms silene arenosa competitive inhibition

Novel ligand-based docking; molecular dynamic simulations;and absorption, distribution, metabolism, and excretion approach to analyzing potential acetylcholinesterase inhibitors for Alzheimer's disease

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Journal of Pharmaceutical Analysis 2018年第6期8卷 413-420页

作者： Subramaniyan Vijayakumar Palani Manogar Srinivasan Prabhu Ram Avadhar Sanjeevkumar Singh Computational Phytochemistry Laboratory PG and Research Department of Botany and Microbiology AVVM Sri Pushpam College (Autonomous) Poondi Computer Aided Drug Design and Molecular Modeling Laboratory Department of Bioinformatics Alagappa University

acetylcholinesterase(AChE) plays an important role in Alzheimer's disease(AD). The excessive activity of AChE causes various neuronal problems, particularly dementia and neuronal cell deaths. Generally, antiAChE drugs induce some serious neuronal side effects in humans. Therefore, this study sought to identify alternative drug molecules from natural products with fewer side effects than those of conventional drugs for treating AD. To achieve this, we developed computational methods for predicting drug and target binding affinities using the Schrodinger suite. The target and ligand molecules were retrieved from established databases. The target enzyme has 539 amino acid residues in its sequence alignment. Ligand molecules of 20 bioactive molecules were obtained from different kinds of plants, after which we performed critical analyses such as molecular docking; molecular dynamic(MD) simulations; and absorption, distribution, metabolism, and excretion(ADME) analysis. In the docking studies, the natural compound rutin showed a superior docking score of à 12.335 with a good binding energy value ofà73.313 kcal/mol. Based on these findings, rutin and the target complex was used to perform MD simulations to analyze rutin stability at 30 ns. In conclusion, our study demonstrates that rutin is a superior drug candidate for AD. Therefore, we propose that this molecule is worth further investigation using in vitro studies.

关键词： Alzheimer's disease acetylcholinesterase Phytocompounds Molecular docking Free energy calculations Molecular dynamic simulations

Crystal structure of acetylcholinesterase catalytic subunits of the malaria vector Anopheles gambiae

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Insect Science 2018年第4期25卷 721-724页

作者： Qian Han Dawn M. Wong Howard Robinson Haizhen Ding Polo C. H. Lam Maxim M. Totrov Paul R. Carlier Jianyong Li Laboratory of Tropical Veterinary Medicine and Vector Biology and Hainan Key Laboratory of Sustainable Utilization of Tropical Bioresources College of Agriculture Hainan University Haikou China Department of Biochemistry Virginia Tech Blacksburg Virginia USA Department of Chemistry Virginia Tech Blacksburg Virginia USA Biology Department Brookhaven National Laboratory Upton New York USA Molsoft LLC San Diego California USA

Dear Editor, acetylcholinesterase （ACHE） hydrolyzes the neurotransmitter acetylcholine at cholinergic synapses in the central nervous system （Toutant, 1989）. Inhibition of the enzyme in insects could lead to the d... 详细信息

关键词： acetylcholinesterase ANOPHELES gambiae CRYSTAL structure

New diterpenoids isolated from Leonurus japonicus and their acetylcholinesterase inhibitory activity

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Chinese Journal of Natural Medicines 2017年第11期15卷 860-864页

作者： WU Han-Kui SUN Ting ZHAO Feng ZHANG Li-Ping LI Gang ZHANG Jie College of Chemistry and Chemical Engineering Anyang Normal University Key Laboratory of Natural Medicine and Immuno-Engineering Henan University

Three new labdane diterpenoids, leojaponicone A(1), isoleojaponicone A(2) and methylisoleojaponicone A(3), were isolated from the herb of Leonurus japonicus. The chemical structures of these secondary metabolites were elucidated on the basis of 1 D and 2 D NMR, including HMQC, and HMBC spectroscopic techniques. All the new compounds were tested in vitro for their acetylcholinesterase and α-glucosidase inhibitory activity. Compounds 1-3 exhibited low inhibitory effects on α-glucosidase with respect to acarbose and exhibited high inhibitory effects on acetylcholinesterase with respect to huperzine A.

关键词： Leonurus japonicus Leojaponicone acetylcholinesterase Labdane diterpenoid