Molecular Docking Study of Phytosterols in Lygodium microphyllum Towards SIRT1 and AMPK, the in vitro Brine Shrimp Toxicity Test, and the Phenols and Sterols Levels in the Extract

Putri Anggreini; Hadi Kuncoro; Sri Adi Sumiwi; Jutti Levita

doi:10.2147/JEP.S438435

Molecular Docking Study of Phytosterols in Lygodium microphyllum Towards SIRT1 and AMPK, the in vitro Brine Shrimp Toxicity Test, and the Phenols and Sterols Levels in the Extract

J Exp Pharmacol. 2023 Dec 22:15:513-527. doi: 10.2147/JEP.S438435. eCollection 2023.

Authors

Putri Anggreini^#^{1

2}, Hadi Kuncoro^#², Sri Adi Sumiwi^#³, Jutti Levita^#³

Affiliations

¹ Faculty of Pharmacy, Padjadjaran University, Sumedang, 46363, Indonesia.
² Faculty of Pharmacy, Mulawarman University, Samarinda, 75119, Indonesia.
³ Department of Pharmacology and Clinical Pharmacy, Faculty of Pharmacy, Padjadjaran University, Sumedang, 46363, Indonesia.

^# Contributed equally.

Abstract

Background: Lygodium microphyllum is a fern plant with various pharmacological activities, and phytosterols were reported contained in the n-hexane and ethyl acetate extract of this plant. Phytosterols are known to inhibit steatosis, oxidative stress, and inflammation. Sirtuin 1 (SIRT1) and adenosine monophosphate-activated protein kinase (AMPK) are the key proteins that control lipogenesis. However, information about L. microphyllum on SIRT1 and AMPK is still lacking.

Purpose: This study aims to investigate the binding mode of phytosterols in L. microphyllum extract towards AMPK and SIRT1, and the toxicity of the extract against brine shrimp (Artemia salina) larvae, and to determine the phenols and sterols levels in the extract.

Methods: The molecular docking was performed towards SIRT1 and AMPK using AutoDock v4.2.6, the toxicity of the extract was assayed against brine shrimp (Artemia salina) larvae, and the phytosterols were analyzed by employing a thin layer chromatography densitometry, and the total phenols were by spectrophotometry.

Results: The molecular docking study revealed that β-sitosterol and stigmasterol could occupy the active allosteric-binding site of SIRT1 and AMPK by binding to important residues similar to the protein's activators. The cold extraction of the plant yields 15.86% w/w. Phytochemical screening revealed the presence of phenols, steroids, flavonoids, alkaloids, and saponins. The total phenols are equivalent to 126 mg gallic acid (GAE)/g dry extract, the total sterols are 954.04 µg/g, and the β-sitosterol level is 283.55 µg/g. The LC₅₀ value of the extract towards A. salina larvae is 203.704 ppm.

Conclusion: Lygodium microphyllum extract may have the potential to be further explored for its pharmacology activities, particularly in the discovery of plant-based anti-dyslipidemic drug candidates. However, further studies are needed to confirm their roles in alleviating lipid disorders.

Keywords: anti-dyslipidemia; cholesterol; inflammation; lipogenesis; phytosterols.

Grants and funding

This research is funded by the Directorate of Higher Education of the Ministry of Education and Culture via the Doctoral Dissertation Grant no. 044/E5/PG.02.00.PL/2023 and 031/E5/PG.02.00.PL/2023. The APC is funded by Padjadjaran University via the Directorate of Research and Community Engagement.