OBJECTIVETo screen the peptide inhibitor of acetylcholinesterase (AChE) from 12-mer random phage display peptide library.

METHODSHuman AChE was used as the target to screen its binding peptides from 12-mer random phage display peptide library. The positive phage clones were isolated after three rounds of biopanning followed then by sequence analysis and their activity evaluation.

RESULTSSix positive phage clones binding to human AChE were obtained, and 4 of them sharing the conservative sequence W(S/P)HY inhibited the enzyme activity of AChE.

CONCLUSIONAcquisition of AChE inhibitor from phage display library provides clues for designing peptide inhibitors of AChE.

Acetylcholinesterase ; metabolism ; Cholinesterase Inhibitors ; metabolism ; pharmacology ; Humans ; Peptide Library ; Peptides ; metabolism ; pharmacology ; Protein Binding

Acetylcholinesterase ; metabolism ; Adult ; Erythrocyte Count ; Erythrocytes ; enzymology ; Humans ; Male ; Motion Sickness ; metabolism ; physiopathology ; Naval Medicine

The chemical constituents from the roots of Aconitum kongboense were studied. Twenty-five diterpenoid alkaloids were isolated from the 95% methanol extract of the roots of A. kongboense by silica gel, reverse-phase silica gel and basic alumina column chromatography. They included a new aconitine-type diterpenoid alkaloid, named as kongboensenine(1), and twenty-four known ones(2-25), i.e., acotarine F(2), acotarine G(3), 14-acetyltalatisamine(4), talatisamine(5), indaconitine(6), yunaconitine(7), chasmanine(8), 6-epi-foresticine(9), homochasmanine(10), 8-deacetyl-yunaconitine(11), chasmaconitine(12), ajaconine(13), franchetine(14), ezochasmanine(15), crassicautine(16), 14-O-deacylcrassicausine(17), genicunine A(18), falconeridine(19), sachaconitine(20), liljestrandisine(21), 8-methyl-14-acetyltalatisamine(22), kongboendine(23), 14-benzoylchasmanine(24) and pseudaconine(25). Their structures were elucidated by common spectroscopic methods including high-resolution electrospray ionization mass spectrometry(HR-ESI-MS) and nuclear magnetic resonance(NMR) techniques. Compounds 2-4, 10, 13, 15-19 and 21-22 were isolated from this plant for the first time. Experimental results showed that all compounds did not have a significant inhibitory activity against acetylcholinesterase(AChE).
Acetylcholinesterase ; Aconitum/metabolism* ; Alkaloids ; Diterpenes ; Magnetic Resonance Spectroscopy ; Molecular Structure ; Plant Roots/metabolism*

Multi-target drugs attract increasing attentions for the therapy of complicated neurodegenerative diseases. In this study, a computer-assisted strategy was applied to search for multi-target compounds by the pharmacophore matching. This strategy has been successfully used to design dual-target inhibitor models against both the acetylcholinesterase (AChE) and poly (ADP-ribose) polymerase-1 (PARP-1). Based on two pharmacophore models matching and physicochemical properties filtering, one hit was identified which could inhibit AChE with IC50 value of (0.337 +/- 0.052) micromol x L(-1) and PARP-1 by 24.6% at 1 micromol x L(-1).
Acetylcholinesterase ; metabolism ; Cholinesterase Inhibitors ; pharmacology ; Computer-Aided Design ; Drug Discovery ; methods ; Poly(ADP-ribose) Polymerase Inhibitors

Objective To study the time-dependent expression and distribution of acetylcholinesterase （AChE） during skin incised wound healing in mice, and discuss its effect in wound healing as well as the feasibility of using it as a reference index for wound age estimation. Methods A total of 45 C57BL/KsJ mice were randomly divided into one control group and eight incised groups. The skin incised wound model was established in the incised groups with samples of skin wounds taken at 6 h, 12 h, 1 d, 3 d, 5 d, 7 d, 10 d and 14 d post-injury respectively, while the uninjured skin tissue was extracted in the control group. Expression and distribution of AChE in skin samples were detected by immunohistochemistry, double immunofluorescence and Western blotting. Results Immunohistochemistry results indicated that AChE was mainly detected in infiltrating polymorphonuclear cells （PMNs） 6 to 12 h post-injury. A large number of AChE-positive mononuclear cells （MNCs） were observed 1 to 3 d post-injury. The AChE-positive cells were mainly fibroblastic cells （FBCs） 5 to 14 d post-injury. The ratio of the AChE-positive cells increased initially 6 h post-injury, and reached the peak at 1 d post-injury. Double immunofluorescent staining showed that the majority of AChE-positive MNCs and FBCs expressed macrophage marker and myofibroblast marker, respectively. Western blotting results showed that the relative expression level of AChE in the incised group was higher than that in the control group averagely, reached the peak at 1 d post-injury, then reached a second peak at 7 d post-injury. Conclusion The expression of AChE is found in PMNs, macrophages and myofibroblast during skin wound healing, which indicates it might be involved in the adjustment of inflammatory response and fibrotic repair after injury. Moreover, combined use of various methods for the detection of the expression of AChE would provide reference for skin wound age estimation.
Acetylcholinesterase/metabolism* ; Animals ; Mice ; Mice, Inbred C57BL ; Skin/pathology* ; Time Factors ; Wound Healing/physiology*

The purpose of the present study was to investigate the effects of cornel iridoid glycoside (CIG) on the activity of cholinesterases in vitro, and to investigate the mechanism of CIG's treating Alzheimer's disease (AD). The sources of cholinesterases were prepared from human blood cells, rat brain homogenate and human blood plasma, respectively. The biochemical methods were used to detect the activity of acetylcholine esterase (AChE) and butyryl cholinesterase (BuChE) to investigate the influence of CIG on cholinesterases. The results showed that CIG inhibited the activity of AChE of human blood cells and rat brain homogenate, with the 50% inhibition rate (IC50) of 1.6 g . L-1 and 3.3 g . L-1, respectively; and the inhibition of AChE of CIG is reversible. CIG also inhibited the activity of BuChE of human blood plasma, with the IC50 of 2.9 g . L-1. In conclusion, CIG can inhibit the activity of AChE and BuChE in vitro, which may be one of the mechanisms of CIG to treat AD.
Acetylcholinesterase ; metabolism ; Animals ; Brain ; drug effects ; metabolism ; Cholinesterase Inhibitors ; pharmacology ; Cholinesterases ; metabolism ; Humans ; Iridoid Glycosides ; pharmacology ; Plasma ; enzymology ; Rats

In order to comprehensively understand current situation about studies on channels and to open train of thought for the study, the relative literatures are analyzed and summarized from morphology, biophysics, biochemistry and so on. Probe into existing models of channel lines from nerves, blood vessels and nerves, gap junction, connective tissue; summarize biophysical characte-ristics of channels from temperature, loose connective tissue, biopressure effect, coherence electromagnetic field, etc.; and summarize biochemical characteristics of channels from ion concentration, exhalant of CO2, extracellular matrix and enzymology on the channel lines. Although the studies have not revealed essence of channel lines, these results reflect survey of studies on tissue structures and biological characteristics of channel lines at present.
Acetylcholinesterase ; metabolism ; Animals ; Body Temperature Regulation ; Calcium ; metabolism ; Carbon Dioxide ; metabolism ; Connective Tissue ; anatomy & histology ; Humans ; Medicine, Chinese Traditional ; Meridians

OBJECTIVETo investigate the diversity of acetylcholinesterase (AChE) activity in variety classes and strains of Culex pipiens pallens and provide a basis for the insecticide-resistance detection of mosquito by biochemical method.

METHODSAChE insensitivity of single mosquito was determined, using acetythiocholine iodide (ATch) as the substrate, 5,5'-dithio-bis (2-nitrobenzoic acid) (DTNB) as the developer, and propoxur as the inhibitor.

RESULTThere were significant differences in AChE activity among the four types of IV instar larvae and 3-day-old adult female of sensitive strain mosquito (P<0.01). The AChE activity of the 3-day-old adult female was higher than that of IV instar larvae of the four types of sensitive strain mosquito (P<0.01). The AChE activity of anti-DDVP (Rd) and anti-propoxur (Rp) strains of Culex pipiens pallens was significantly higher than that of sensitive (S) strain (P<0.01), while the AChE activity of anti-cypermethrin (Rc) strain of Culex pipiens pallens was similar to that of S strain (P>0.05). The individual frequency of insensitive AChE of Rd and Rp strains of Culex pipiens pallens was significantly higher than that of sensitive (S) strain (P<0.01), while the individual frequency of insensitive AChE of Rc strain of Culex pipiens pallens was similar to that of S strain(P>0.05).

CONCLUSIONThe AChE activity determination can be used to examine the insecticide-resistance of mosquito.

Acetylcholinesterase ; metabolism ; Animals ; Culex ; classification ; enzymology ; Dichlorvos ; pharmacology ; Female ; Insecticide Resistance ; Propoxur ; pharmacology ; Pyrethrins ; pharmacology ; Species Specificity

A series of novel 2-amino-4-phenylthiazole derivatives were designed and synthesized, furthermore, their inhibition effect on acetylcholinesterase were investigated. 2-Amino-4-phenylthiazoles were prepared from alpha-bromoacetophenones by Hantzsch reaction, acylation reaction and substitution reaction. Moreover, their bioactivities as AChE inhibitors in vitro were measured with Ellman spectrophotometry. The results showed that most of them had a certain inhibition activity on AChE, and the compound 8a was the best of them. The IC50 of 8a to AChE is 3.54 micromol x L(-1), and the value was better than that of rivastigmine. 2-Amino-4-phenylthiazole derivatives showed a certain bioactivity in vitro, which were worth further investigation.
Acetylcholinesterase ; metabolism ; Cholinesterase Inhibitors ; chemical synthesis ; chemistry ; pharmacology ; Drug Design ; Inhibitory Concentration 50 ; Molecular Structure ; Thiazoles ; chemical synthesis ; chemistry ; pharmacology

The target compounds were prepared from 5-aminobenzimidazolone by two steps reaction, and their AChE inhibitory activities were measured by Ellman method in vitro. The AChE inhibitory activity of compound 4d is the best of them, and its IC50 value is equal to 7.2 μmol·L(-1), which is better than that of rivastigmine; moreover the 4d had no inhibitory activities to BuChE. Therefore, the inhibitory activities of 5-aminobenzimidazolone derivatives to acetylcholinesterase are worth further researching.
Acetylcholinesterase ; metabolism ; Benzimidazoles ; chemical synthesis ; chemistry ; Cholinesterase Inhibitors ; chemical synthesis ; chemistry ; Drug Design ; Phenylcarbamates ; chemistry ; Rivastigmine ; Structure-Activity Relationship