Antioxidant, Antibacterial, Antidiabetic Potential, and In silico Analysis of Rhus chinensis from Western Nepal

Abstract

Background: Rhus chinensis Mill, indigenous wild fruit primarily found in the hilly region of Nepal. The ripe fruit is very sour and considered medicinal as a remedy for colic pain. In addition, their astringent and styptic qualities are used internally to treat illnesses such as diarrhea and hemorrhage. Also, they are used as a common component of polyherbal medications for diabetic mellitus.

Objectives: This work aimed to determine the total phenolic and flavonoid content, antioxidant, antibacterial, α-glucosidase, and α-amylase inhibition activity of the crude extract and fractions of Rhus chinensis Mill. Additionally, molecular docking of compounds from Rhus chinensis was performed.

Methods: Folin Ciocalteu's (FC) reagent was used to estimate total phenolic content. Likewise, the aluminium trichloride method was applied to determine total flavonoid content. A 2,2-diphenyl-1- picrylhydrazyl (DPPH) free radical scavenging assay was performed for the antioxidant activity. Furthermore, the substrate-based enzyme inhibition assay was carried out for α-glucosidase and α- amylase inhibition activity of R. chinensis. P-nitrophenyl-α-D-glucopyranoside (PNPG) and 2- Chloro-4-Nitrophenyl-α-D-Maltotrioside (CNPG3) were used as substrates for α-glucosidase and α- amylase inhibition assay, respectively. Similarly, the well-diffusion method was used for the antibacterial activity. Autodock vina was used to perform molecular docking.

Results: The total phenolic and flavonoid content of R. chinensis fruit were 117.092±1.1 mg GAE/g and 62.41±1.23 mg QE/g, respectively. The IC₅₀ value for antioxidant activity of the crude extract and its fractions ranged from 3.12±1.15μg/mL to 50.85±2.10μg/mL. Similarly, the IC₅₀ for α- glucosidase inhibition ranged from2.33±1.01μg/mL to 28.34±2.79μg/mL. Likewise, The IC₅₀ of R. chinensis crude methanolic extract against α-amylase was 120.3±1.382μg/mL. The antibacterial activity of R. chinensis was effective against gram-positive bacteria; Staphylococcus aureus (ZOI=11.0) and Bacillus subtilis (ZOI=9.0). Quercetin-3-O-rhamnoside and Myricetin-3-Orhamnoside showed excellent binding to the active site of protein with binding energy -9.4kcal/mol and -9.6kcal/mol, respectively.

Conclusion: Rhus chinensis Mill is a potent antioxidant and inhibits enzymes; α-glucosidase and α- amylase. In addition, the methanolic extract of this plant shows antibacterial activity. However, further research is required to determine the inhibiting compounds.

Keywords: Antibacterial; Antioxidant; Molecular docking.; Rhus chinensis; α-Amylase inhibition; α-Glucosidase inhibition.