Abstract
BackgroundWedelia chinensis has been reported as a folk medicine for the treatment of different diseases including neurodegenerative disease. Although the plant has been studied well for diverse biological activities, the effect of this plant in neurological disorder is largely unknown. The present study was undertaken to evaluate the cholinesterase inhibitory and antioxidant potential of W. chinensis.MethodsThe extract and fractions of the plant were evaluated for acetylcholinesterase and butyrylcholinesterase inhibitory activity by modified Ellman method. The antioxidant activity was assessed in several in vitro models/assays such as reducing power, total antioxidant capacity, total phenolic and flavonoid content, scavenging of 2,2′-diphenyl-1-picrylhydrazyl (DPPH) free radical and hydroxyl radical, and inhibition of brain lipid peroxidation. Chromatographic and spectroscopic methods were used to isolate and identify the active compound from the extract.ResultsAmong the fractions, aqueous fraction (AQF) and ethylacetate fraction (EAF) exhibited high inhibition against acetylcholinesterase (IC50: 40.02 ± 0.16 μg/ml and 57.76 ± 0.37 μg/ml) and butyrylcholinesterase (IC50: 31.79 ± 0.18 μg/ml and 48.41 ± 0.05 μg/ml). Similarly, the EAF and AQF had high content of phenolics and flavonoids and possess strong antioxidant activity in several antioxidant assays including DPPH and hydroxyl radical scavenging, reducing power and total antioxidant activity. They effectively inhibited the peroxidation of brain lipid in vitro with IC50 values of 45.20 ± 0.10 μg/ml and 25.53 ± 0.04 μg/ml, respectively. A significant correlation was observed between total flavonoids and antioxidant and cholinesterase inhibitory activity. Activity guided chromatographic separation led to the isolation of a major active compound from the EAF and its structure was elucidated as apigenin by spectral analysis.ConclusionsThe potential ability of W. chinensis to inhibit the cholinesterase activity and peroxidation of lipids suggest that the plant might be useful for the management of AD.
Highlights
Wedelia chinensis has been reported as a folk medicine for the treatment of different diseases including neurodegenerative disease
Chemicals Donepezil (CAS 120011–70-3), galantamine (CAS 195304-4), acetylthiocholine iodide (CAS 1866-15-5) and Sbutyrylthiocholine iodide (CAS 1866-16-6), DPPH (2,2′diphenyl-1-picrylhydrazyl) (CAS 1898-66-4), ammonium molybdate, Folin-Ciocalteu reagent, thiobarbituric acid (TBA) (CAS 504–17-6), tricholoroacetic acid (TCA), 2deoxy-D-ribose (CAS 533–67-5), 5,5′-dithio-bis-(2-nitro) benzoic acid (DTNB) (CAS 69–78-3), triton X-100, aluminum chloride, potassium ferricyanide, and TrisHCl were procured from Sigma-Aldrich, Germany
Qualitative analysis of the four fractions showed that they all contained phenolics and flavonoids, but higher amounts were found in the ethylacetate fraction (EAF) and aqueous fraction (AQF) (Supplementary Table S1)
Summary
Chemicals Donepezil (CAS 120011–70-3), galantamine (CAS 195304-4), acetylthiocholine iodide (CAS 1866-15-5) and Sbutyrylthiocholine iodide (CAS 1866-16-6), DPPH (2,2′diphenyl-1-picrylhydrazyl) (CAS 1898-66-4), ammonium molybdate, Folin-Ciocalteu reagent, thiobarbituric acid (TBA) (CAS 504–17-6), tricholoroacetic acid (TCA), 2deoxy-D-ribose (CAS 533–67-5), 5,5′-dithio-bis-(2-nitro) benzoic acid (DTNB) (CAS 69–78-3), triton X-100, aluminum chloride, potassium ferricyanide, and TrisHCl were procured from Sigma-Aldrich, Germany. Plant extract or reference compound (6.25–100 μg/ml concentration) was added to a 1 ml reaction mixture containing 2.8 mM 2deoxy-2-ribose, 20 mM phosphate buffer (pH 7.4), 100 μM FeCl3, 100 μM EDTA, 1 mM H2O2 and 100 μM ascorbic acid and incubated at 37 °C for 60 min. Plant extract or reference compound (12.5–200 μg/ ml concentration) was added in an enzyme solution and incubated at 37 °C for 15 min for interaction. This was followed by the addition of a 50 mM sodium phosphate buffer (pH 8.0) containing 0.5 mM acetylthiocholine and 1 mM DTNB and immediately the absorbance of the solution was recorded against a blank solution.
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