Research Article | Volume: 15, Issue: 7, July, 2025

Oyster mushroom ethanolic extract promotes lipid accumulation and upregulates Cebpa and Slc2a4 gene expression while reducing Slc2a4 promoter methylation during 3T3-L1 adipocyte differentiation

Eko Fuji Ariyanto,  Devina Nabila Fitri,  Seysa Azima Arvieri,  Sylvia Mahaedally,  Erlina Widiarsih,  Nurul Qomarilla,  Nurul Setia Rahayu,  Tenny Putri Wikayani,  Henhen Heryaman,  Afiat Berbudi,  Enny Rohmawaty,  Dwi Wahyudha Wira,  Muhammad Hasan Bashari,  Yunisa Pamela,  Yuni Susanti Pratiwi,  Mohammad Ghozali   

Eko Fuji Ariyanto1, Devina Nabila Fitri2, Seysa Azima Arvieri2, Sylvia Mahaedally2, Erlina Widiarsih3, Nurul Qomarilla4, Nurul Setia Rahayu3, Tenny Putri Wikayani4, Henhen Heryaman1, Afiat Berbudi1, Enny Rohmawaty1, Dwi Wahyudha Wira1, Muhammad Hasan Bashari1, Yunisa Pamela1, Yuni Susanti Pratiwi1, Mohammad Ghozali1

1Department of Biomedical Sciences, Faculty of Medicine, Universitas Padjadjaran, Sumedang, Indonesia.

2Faculty of Medicine, Universitas Padjadjaran, Sumedang, Indonesia.

3Molecular Genetics Laboratory, Faculty of Medicine, Universitas Padjadjaran, Bandung, Indonesia.

4Cell Culture Laboratory, Faculty of Medicine, Universitas Padjadjaran, Bandung, Indonesia.

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Published:  Jun 05, 2025

DOI: 10.7324/JAPS.2025.231795
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Abstract

This study assessed the effect of an oyster mushroom ethanolic extract on lipid accumulation, Cebpa and Slc2a4 gene expression, and methylation level on Slc2a4 promoter during 3T3-L1 adipocyte differentiation. The oyster mushrooms were extracted using the maceration method with ethanol. 3T3-L1 preadipocytes were cultured and differentiated using methylisobutylxanthine, dexamethasone, and insulin cocktail-containing DMEM. Twenty-four hours after seeding, the cells were incubated with 25, 50, or 100 μg/ml of the oyster mushroom ethanolic extract for 12 days. The media were replaced every 2 days. On day 12 after differentiation induction, oil red O staining and quantitative RT-PCR were performed to analyze lipid accumulation and to measure the mRNA expression of Cebpa and Slc2a4, respectively. The methylation level on the Slc2a4 promoter was measured using pyrosequencing of bisulfite-treated DNA samples. Treatment with the oyster mushroom ethanolic extract increased the mRNA expression of Cebpa and Slc2a4 in a dose-dependent manner. The highest expression of Cebpa and Slc2a4 was observed with the addition of the 50-μg/ml oyster mushroom ethanolic extract (about two-fold and fifty-fold higher than that of the control, respectively). Moreover, lipid accumulation increased after the addition of the oyster mushroom ethanolic extract. Interestingly, the oyster mushroom ethanolic extract slightly reduced the Slc2a4 methylation level on one of the CpG sites analyzed, in a dose-dependent manner. The oyster mushroom ethanolic extract increased the lipid accumulation and the Cebpa and Slc2a4 expression, while reducing the methylation level in the Slc2a4 gene promoter region.


Keyword:     3T3-L1 cell line Cebpa epigenetics oyster mushroom Slc2a4 type 2 diabetes mellitus

Citation:

Ariyanto EF, Fitri DN, Arvieri SA, Mahaedally S, Widiarsih E, Qomarilla N, Rahayu NS, Wikayani TP, Heryaman H, Berbudi A, Rohmawaty E, Wira DW, Bashari MH, Pamela Y, Pratiwi YS, Ghozali M. Oyster mushroom ethanolic extract promotes lipid accumulation and upregulates Cebpa and Slc2a4 gene expression while reducing Slc2a4 promoter methylation during 3T3-L1 adipocyte differentiation. J Appl Pharm Sci. 2025;15(07):242–252. http://doi.org/10.7324/JAPS.2025.231795

Copyright: © The Author(s). This is an open-access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
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