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ORIGINAL ARTICLE
Year : 2018  |  Volume : 14  |  Issue : 57  |  Page : 448-452

In vitro anticholinesterase and neurotoxicity activities of Ocotea aciphylla fractions


1 Department of Health, Laboratory of Toxicology, State University of Feira de Santana, Feira de Santana; Department of Biochemistry and Biophysics, Institute of Health Sciences, Laboratory of Neurochemistry and Cell Biology, Federal University of Bahia, Salvador, Bahia, Brazil
2 Department of Health, Laboratory of Toxicology, State University of Feira de Santana, Feira de Santana, Bahia, Brazil
3 Department of Health, Laboratory of Phytochemistry, State University of Feira de Santana, Feira de Santana, Bahia, Brazil
4 Department of Biochemistry and Biophysics, Institute of Health Sciences, Laboratory of Neurochemistry and Cell Biology, Federal University of Bahia, Salvador, Bahia, Brazil

Correspondence Address:
Mariana Borges Botura
Departamento de Saude, Laboratorio de Toxicologia, Universidade Estadual de Feira de Santana, 44.036-900 Feira de Santana, Bahia
Brazil
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/pm.pm_366_17

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Background: Ocotea species are known to produce secondary metabolites with a range of biological activities. This study aimed to evaluate the in vitro acetylcholinesterase (AChE) inhibition and neurotoxicity activities of the Ocotea aciphylla leaves. Materials and Methods: The in vitro anticholinesterase effect of crude extracts of O. aciphylla was investigated by means of spectrophotometric microplate assay. The most active extract, aqueous extract (AQE), was fractionated using column chromatography with silica gel as stationary phase to furnish several fractions that were also evaluated for the anticholinesterase effect. The neurotoxicity activity of AQE and active fraction (F9) was investigated in rat adrenal medulla pheochromocytoma strain cultures by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide test. The chemical characterization of the most active fraction was performed through high-performance liquid chromatography coupled with mass spectrometry multistage (HPLC-MS/MS). Results: Ethanolic extract (EE) and AQE exhibited significant inhibitory effects of the activity of AChE, with inhibitory concentration (IC50) of 443.7 and 412.8 μg/mL, respectively. Among the fractions, F9 was more effective AChE inhibition with IC50of 286.2 μg/mL. In the neurocytotoxicity assays, only the F9, at the highest concentration (500 μg/mL), induced a significant reduction in cell viability. HPLC-MS/MS analysis of the active fraction enabled the characterization of the natural compounds, i.e., procyanidin B-type dimer, propelargonidin dimer, catechin, and methoxy-luteolin-deoxyhexose-hexose. Conclusion: The leaves of O. aciphylla showed in vitro anticholinesterase activity and low neurotoxicity, and these effects might be related to the presence of phenolic compounds. Abbreviation used: AChE: acetylcholinesterase; ACTI: Acetylthiocholine iodide; AD: Alzheimer's disease; AQE: Aqueous extract; BSA: Bovine serum albumin; DMSO: Dimethyl sulfoxide; DTNB: 5,5'-dithiobis (2-nitrobenzoic acid); EAE: Ethyl acetate extract; EE: Ethanolic extract; EtOAc: Ethyl acetate; HE: Hexane extract; Hex: Hexane; HPLC-MS/MS: High-performance liquid chromatography coupled with mass spectrometry multistage; MeOH: Methanol; MTT: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide; PC-12: Rat adrenal medulla pheochromocytoma strain; RPMI: Roswell park memorial institute medium.


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