[ Objective ] The paper was to systemically study the characteristics of acetyl cholinesterase （ACHE） in Pardosa astrigera, and confirm the occurrence and development of its pesticide resistance and the resistance l...[ Objective ] The paper was to systemically study the characteristics of acetyl cholinesterase （ACHE） in Pardosa astrigera, and confirm the occurrence and development of its pesticide resistance and the resistance level, thereby establishing a quick and accurate detection method for enzyme activity. [ Method] The optimal conditions for assaying the activity of AChE in different parts of P. astrigera were determined by orthogonal experiment. The distribution conditions of acetyl cholinesterase （AChE） in different tissues of P. astrigera were further studied, and the sensitivities of the enzyme tO four common pesticides were also determined. [ Result] The optimal condition for assaying the activity of AChE in cephalothorax, abdomen and appendage of P. astrigera was as follows, enzyme concentrations： 12, 18 and 29 g/L; substrate concentrations： 0.6, 1.0 and 1.0 mmol/L; pH value, 7.0; reaction temperatures： 30, 35, 35℃ ; reaction time, 5 rain. AChE was mainly distributed in the cephalothorax of P. astrigera. The specific activity of AChE in the extract solution with Triton X-100 was higher than that in the solution without Triton X-100. The median inhibitory concentrations （IC50） of methomyl, phoxim, betacypermethrin, chlorpyrifos against AChE in the cephalothorax of P. astrigera were 7.76 × 10^-5, 1.76×10^-4, 4.12 ×10^-4 and 4.94 ×10^-4 mol/L, respectively. [ Conclusion] AChEs in P. astrigera were membrane-bounded. The inhibition of four pesticides against AChE in the cephalothorax of P. astrigera had good dese-effect, this indicated that AChE in the cephalothorax of P. astrig- era could be used as the biochemical marker to monitor the contamination of organic phosphorus, carbamate and pyrethroid pesticides in the environment.展开更多
To find promising new multitargeted AD （Alzheimer＇s disease） inhibitors, the 3D-QSAR （three-dimensional quantitative structure-activity relationship） model for 32 AD inhibitors was established by using the CoMFA ...To find promising new multitargeted AD （Alzheimer＇s disease） inhibitors, the 3D-QSAR （three-dimensional quantitative structure-activity relationship） model for 32 AD inhibitors was established by using the CoMFA （comparative molecular field analysis） and CoMSIA （comparative molecular similarity index analysis） methods. Results showed that the CoMFA and CoMSIA models were constructed successfully with a good cross-validated coefficient （q2） and a non-cross-validated coefficient （R2）, and the binding modes obtained by molecular docking were in agreement with the 3D-QSAR results, which suggests that the present 3D-QSAR model has good predictive capability to guide the design and structural modification of novel multitargeted AD inhibitors. Meanwhile, we found that one side of inhibitory molecule should be small group so that it would be conductive to enter the gorge to interact with the catalytic active sites of AChE （acetylcholinesterase）, and the other side of inhibitory molecule should be large group so that it would be favorable for interaction with the peripheral anionic site of ACHE. Furthermore, based on the 3D-QSAR model and the binding modes of AChE and [3-secretase （BACE-1）, the designed molecules could both act on dual binding sites of AChE （catalytic and peripheral sites） and dual targets （ACHE and BACE-1）. We hope that our results could provide hints for the design of new multitargeted AD derivatives with more potency and selective activity.展开更多
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 d...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.展开更多
The inhibition of acetylcholinesterase (ACHE) activity has been widely used as a biomarker in an animal exposed to the pesticides. However, the interaction of extensively used organocarbamate insecticide, carbofuran, ...The inhibition of acetylcholinesterase (ACHE) activity has been widely used as a biomarker in an animal exposed to the pesticides. However, the interaction of extensively used organocarbamate insecticide, carbofuran, with the nervous system of the aquatic organisms is not properly studied. AChE is a key enzyme which catalyses the hydrolysis of acetylcholine, a neurotransmitter at the neuromuscular junctions, and thus regulates the neurotransmission system. In the present study, we have evaluated the impact of sub-acute concentrations (0.01 and 0.02 mg/L i.e. 1/20th and 1/10th of LC50) of carbofuran on the activity of acetylcholinesterase,from different tissues of Clarias batrachus, a fresh water teleost, after 96 hr and 15 days exposure periods in vivo. The carbofuran significantly reduced the activity of AChE in different tissues of C. batrachus at both concentrations and periods of exposure. The greater inhibition of AChE activities were recorded in fish tissues at higher carbofuran concentration (0.02 mg/L) after longer (15days) treatment period. The inhibition of AChE activity in all fish tissues tested was dependent on pesticide concentration and the duration of treatment. AChE from the tissues of C. batrachus was found to be a true cholinesterase as it was completely inhibited by the small concentration (nM) of eserine as tested in vitro. It was found that carbofuran at very low concentration exerted significant inhibitory effect on AChE activity in fish tissues.展开更多
基金Supported by Shanxi Scholarship Fund(2009042)Youth Science andTechnology Research Fund in Shanxi Province(2008021041)~~
文摘[ Objective ] The paper was to systemically study the characteristics of acetyl cholinesterase （ACHE） in Pardosa astrigera, and confirm the occurrence and development of its pesticide resistance and the resistance level, thereby establishing a quick and accurate detection method for enzyme activity. [ Method] The optimal conditions for assaying the activity of AChE in different parts of P. astrigera were determined by orthogonal experiment. The distribution conditions of acetyl cholinesterase （AChE） in different tissues of P. astrigera were further studied, and the sensitivities of the enzyme tO four common pesticides were also determined. [ Result] The optimal condition for assaying the activity of AChE in cephalothorax, abdomen and appendage of P. astrigera was as follows, enzyme concentrations： 12, 18 and 29 g/L; substrate concentrations： 0.6, 1.0 and 1.0 mmol/L; pH value, 7.0; reaction temperatures： 30, 35, 35℃ ; reaction time, 5 rain. AChE was mainly distributed in the cephalothorax of P. astrigera. The specific activity of AChE in the extract solution with Triton X-100 was higher than that in the solution without Triton X-100. The median inhibitory concentrations （IC50） of methomyl, phoxim, betacypermethrin, chlorpyrifos against AChE in the cephalothorax of P. astrigera were 7.76 × 10^-5, 1.76×10^-4, 4.12 ×10^-4 and 4.94 ×10^-4 mol/L, respectively. [ Conclusion] AChEs in P. astrigera were membrane-bounded. The inhibition of four pesticides against AChE in the cephalothorax of P. astrigera had good dese-effect, this indicated that AChE in the cephalothorax of P. astrig- era could be used as the biochemical marker to monitor the contamination of organic phosphorus, carbamate and pyrethroid pesticides in the environment.
基金Acknowledgments The authors acknowledge the financial support of the Natural Science Foundation of Guangxi Province （No. 2013GXNSFAA019019） and the Natural Science Foundation of Guangxi Province （No. 2013GXNSFAA019041）.
文摘To find promising new multitargeted AD （Alzheimer＇s disease） inhibitors, the 3D-QSAR （three-dimensional quantitative structure-activity relationship） model for 32 AD inhibitors was established by using the CoMFA （comparative molecular field analysis） and CoMSIA （comparative molecular similarity index analysis） methods. Results showed that the CoMFA and CoMSIA models were constructed successfully with a good cross-validated coefficient （q2） and a non-cross-validated coefficient （R2）, and the binding modes obtained by molecular docking were in agreement with the 3D-QSAR results, which suggests that the present 3D-QSAR model has good predictive capability to guide the design and structural modification of novel multitargeted AD inhibitors. Meanwhile, we found that one side of inhibitory molecule should be small group so that it would be conductive to enter the gorge to interact with the catalytic active sites of AChE （acetylcholinesterase）, and the other side of inhibitory molecule should be large group so that it would be favorable for interaction with the peripheral anionic site of ACHE. Furthermore, based on the 3D-QSAR model and the binding modes of AChE and [3-secretase （BACE-1）, the designed molecules could both act on dual binding sites of AChE （catalytic and peripheral sites） and dual targets （ACHE and BACE-1）. We hope that our results could provide hints for the design of new multitargeted AD derivatives with more potency and selective activity.
基金DST-SERB (SB/YS/LS-109/2014) for providing financial assistance for this project
文摘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.
文摘The inhibition of acetylcholinesterase (ACHE) activity has been widely used as a biomarker in an animal exposed to the pesticides. However, the interaction of extensively used organocarbamate insecticide, carbofuran, with the nervous system of the aquatic organisms is not properly studied. AChE is a key enzyme which catalyses the hydrolysis of acetylcholine, a neurotransmitter at the neuromuscular junctions, and thus regulates the neurotransmission system. In the present study, we have evaluated the impact of sub-acute concentrations (0.01 and 0.02 mg/L i.e. 1/20th and 1/10th of LC50) of carbofuran on the activity of acetylcholinesterase,from different tissues of Clarias batrachus, a fresh water teleost, after 96 hr and 15 days exposure periods in vivo. The carbofuran significantly reduced the activity of AChE in different tissues of C. batrachus at both concentrations and periods of exposure. The greater inhibition of AChE activities were recorded in fish tissues at higher carbofuran concentration (0.02 mg/L) after longer (15days) treatment period. The inhibition of AChE activity in all fish tissues tested was dependent on pesticide concentration and the duration of treatment. AChE from the tissues of C. batrachus was found to be a true cholinesterase as it was completely inhibited by the small concentration (nM) of eserine as tested in vitro. It was found that carbofuran at very low concentration exerted significant inhibitory effect on AChE activity in fish tissues.
