Browsing by Author "Dhiraj Kumar Choudhary"

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Catechin isolated from faba beans (Vicia faba L.): insights from oxidative stress and hypoglycemic effect in yeast cells through confocal microscopy, flow cytometry, and in silico strategy
(Taylor and Francis Ltd., 2022) Dhiraj Kumar Choudhary; Navaneet Chaturvedi; Amit Singh; Abha Mishra
The present aim of this investigation was to evaluate the effect of catechin from faba beans on oxidative stress and glucose uptake in yeast cells. Flow cytometry approach indicated that 2-NBDG (1.98 ± 0.37) seed extract had a lower relative fluorescence signal than methanol (5.98 ± 0.67) and acetone seed extract (4.43 ± 0.55). In comparison to the control and seed extract, H2O2 exposure increased the apoptosis rate of yeast cells from 8.20% to 64.80%. Yeast cells incubated with H2O2 produced significantly more ROS intensity (162 ± 4.32, p < 0.05) than control cells (118 ± 2.52, p < 0.05) and less than seed extract-treated cells. Molecular dynamics simulation studies were performed for cat:α-amylase (catechin-α-amylase complex) which revealed the stable and mixed mode of inhibition during a simulation. The synergistic action of polyphenols or catechin present in seed extract may be responsible for the anti-oxidative stress and hypoglycaemic effects. The findings of this study may provide insight into the further development of a novel antidiabetic drug for T2DM. Communicated by Ramaswamy H. Sarma. © 2021 Informa UK Limited, trading as Taylor & Francis Group.
Characterization, inhibitory activity and mechanism of polyphenols from faba bean (gallic-acid and catechin) on α-glucosidase: insights from molecular docking and simulation study
(Taylor and Francis Inc., 2020) Dhiraj Kumar Choudhary; Navaneet Chaturvedi; Amit Singh; Abha Mishra
The chemo-profiling of ethanolic extract of faba beans seeds was performed and explored as an α-glucosidase inhibitor. The inhibition of α-glucosidase is one of the alternatives approach to control postprandial hyperglycemia by, resulting in the delay of the carbohydrate digestion of absorbable monosaccharides. Ethanolic seed extract showed phenolic compounds, flavonoid such as gallic acid (m/z [M– H] = 169.0124,C7H6O5) ellagic acid derivatives epigallocatechin (m/z [M– H = 305.0644,C15H14O7),catechin (m/z [M– H] = 289.0656,C15H14O6), epigallocatechin gallate (m/z [M– H] = 457.0578,C22H18O11) and epicatechin monogallate (m/z [M– H] = 441.081, C22H18O10). The extract was found to exert inhibitory activity (88.28 ± 2.67%) (IC50 value of 2.30 ± 0.032 mg/mL) with a mixed mode of inhibition (Km, apparent = 0.54 ± 0.020 mM and Vmax, apparent 0.136 ± 0.04 mM/min). Molecular docking studies of gallic acid and catechin on α-glucosidase proposed productive binding modes having binding energy (−6.58 kcal/mol and −7.25 kcal/mol) with an effective number of hydrogen bonds and binding energy. Tyr63, Arg197, Asp198, Glu 233, Asn324, Asp 326 of α-glucosidase participated in binding events with gallic acid and catechin. Molecular dynamics simulation studies were performed for both complexes i.e. gal:α-glucosidase and cat:α-glucosidase along with apo state of α-glucosidase, which revealed stable systems during the simulation. These findings of the present study may give an insight into the further development of the novel antidiabetic drug from the seeds of faba beans. © 2019, © 2019 Taylor & Francis Group, LLC.
Investigation of hypoglycemic effects, oxidative stress potential and xanthine-oxidase activity of polyphenols (gallic acid, catechin) derived from faba bean on 3T3-L1 cell line: Insights into molecular docking and simulation study
(Oxford University Press, 2020) Dhiraj Kumar Choudhary; Navaneet Chaturvedi; Amit Singh; Abha Mishra
Hypoglycemic potential and xanthine-oxidase (XO) activity of polyphenols from faba bean were evaluated in the 3T3-L1 cell line, and an interaction study in silico with XO was performed with considerable bioactive components of acetone extract of faba beans. The protonated and fragmented behavior of acetone seed extract revealed the presence of gallic acid (MS/MS, m/z 169) and catechin (MSn,m/z 288.3). Flow cytometry study explained the effect of hydrogen peroxide (H2O2) on cell line as cell death was increased from 9.72 to 41.66% as compared to the control (without H2O2). The atomic force microscopy (AFM), scanning electron microscopy and reactive oxygen species measurement also confirmed the protective effect of polyphenols in the 3T3-L1 cell lines. Oxidative stress through propidium iodide and 4',6-diamidino-2-phenylindole staining demonstrated that the apoptotic ratio was 0.35 ± 2.62 (P < 0.05) and 30 ± 2.54% in H2O2-treated cells, respectively, as compared to control. The observations of flow cytometry and confocal microscopy marked the effect of seed extract (0.86 ± 0.031, 3.52 ± 0.52, P < 0.05), on glucose uptake in cells through the better relative fluorescence intensity than that of the control. Moreover, molecular docking and molecular dynamics simulation studies gave an insight into the predicted residues that hold favorable polyphenolic-specific interactions. The probable binding modes of the gallic acid and catechin from this study may extend the knowledge of the XO-polyphenol interactions and offered the way to design the analogs of acetone seed extract with reduced toxicity. © 2020 Royal Society of Chemistry. All rights reserved.