β-sitosterol isolated from the leaves of Trema orientalis (Cannabaceae) promotes viability and proliferation of BF-2 cells

Author:

Mekarunothai Amita12,Bacher Markus3,Buathong Raveevatoo4,Intarasam Saraphorn5,Tayana Ngampuk67,Kongkiatpaiboon Sumet67,Charoenrat Theppanya2,Napiroon Tiwtawat27

Affiliation:

1. Program in Applied Biological Sciences, Chulabhorn Graduate Institute, Bangkok, Bangkok, Thailand

2. Department of Biotechnology, Faculty of Science and Technology, Thammasat University, Pathum Thani, Pathum Thani, Thailand

3. Institute of Chemistry of Renewable Resources, University of Natural Resources and Life Sciences Vienna (BOKU), Tulln an der Donau, Tulln an der Donau, Austria

4. Department of Biochemistry, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Bangkok, Thailand

5. Songkhla Aquatic Animal Health Research and Development Center, Department of Fisheries, Ministry of Agriculture and Cooperatives, Songkhla, Songkhla, Thailand

6. Drug Discovery and Development Center, Thammasat University, Pathum Thani, Pathum Thani, Thailand

7. Thammasat University Research Unit in Cannabis and Herbal Products Innovation, Thammasat University, Pathum Thani, Pathum Thani, Thailand

Abstract

Trema orientalis is a pioneer species in the cannabis family (Cannabaceae) that is widely distributed in Thai community forests and forest edges. The mature leaves are predominantly used as an anti-parasite treatment and feed for local freshwater fish, inspiring investigation of their phytochemical composition and bioactivity. The purpose of this work was to investigate the bioactive compounds in T. orientalis leaf extract and their cytotoxicity in the BF-2 fish cell line (ATCC CCL-91). Flash column chromatography was used to produce 25 mL fractions with a mixture solvent system comprised of hexane, diethyl ether, methanol, and acetone. All fractions were profiled with HPLC-DAD (mobile phase methanol:aqueous buffer, 60:40 v/v) and UV detection (wavelengths 256 and 365 nm). After drying, a yellowish powder was isolated from lipophilic leaf extract with a yield of 280 µg/g dry weight. Structure elucidation by nuclear magnetic resonance (NMR) indicated it to consist of pure β-sitosterol. The lipophilic extract and pure compound were evaluated for cytotoxicity using BF-2 cells. MTT assays showed both leaf extract and pure compound at 1 µg/mL to increase cell viability after 24 h treatment. The respective half maximal inhibitory concentration (IC50) values of leaf extract and β-sitosterol were 7,027.13 and 86.42 µg/ml, indicating a lack of toxicity in the BF-2 cell line. Hence, T. orientalis can serve as a source of non-toxic natural lipophilic compounds that can be useful as bioactive ingredients in supplement feed development.

Funder

The Agricultural Research Development Agency (Public Organization) Thailand, (ARDA) which provided research grants in medicinal plants cluster

Publisher

PeerJ

Reference31 articles.

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4. Molecular mechanism of β-sitosterol and its derivatives in tumor progression;Bao;Frontiers in Oncology,2022

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