Aaptamine Inhibits Lipid Accumulation and Pparg and Slc2a4 Expression While Maintaining the Methylation of the Pparg Promoter During 3T3-L1 Adipocyte Differentiation

Eko Fuji Ariyanto,1 Ibnu Wijaya,2 Zaky Alif Pradian,2 Agung Putri Mayasari Bhaskara,2 Mohammad Parezal Lastialno,3 Putri Halleyana Adrikni Rahman,1 Muhammad Hasan Bashari,1 Nandina Oktavia,1 Muhammad Iman Pratama Putra,4 Yuni Susanti Pratiwi,1 Henhen Heryaman,1 Diah Dhianawaty1 1Department of Biomedical Sciences, Faculty of Medicine, Universitas Padjadjaran, Sumedang, Indonesia; 2Undergraduate Program of Medical Doctor, Faculty of Medicine, Universitas Padjadjaran, Sumedang, Indonesia; 3Master Program of Biomedical Sciences, Faculty of Medicine, Universitas Padjadjaran, Bandung, Indonesia; 4Department of Internal Medicine, Faculty of Medicine, Universitas Padjadjaran - Hasan Sadikin General Hospitale, Bandung, IndonesiaCorrespondence: Eko Fuji Ariyanto, Department of Biomedical Sciences, Faculty of Medicine, Universitas Padjadjaran, Sumedang, Indonesia, Tel +6281316099791, Email [email protected]: Excessive adipogenesis plays a role in the development of obesity and related metabolic disorders. Aaptamine is an alkaloid compound that has been proven to have various effects, however, no studies have yet investigated its effects on adipogenesis. This study aims to examine whether aaptamine inhibits lipid accumulation and Pparg and Slc2a4, two important genes in adipogenesis, mRNA expression, and increases the methylation of the Pparg promoter. This study strengthens the insights regarding these genes regulation, with future research potentially expanding to other adipogenic regulators for a broader perspective.Methods: The effects of aaptamine (0 μM, 25 μM and 50 μM) were investigated in 3T3-L1 preadipocytes. The adipocytes were differentiated using a medium containing 3-isobutyl-1-methylxanthine, dexamethasone, and insulin. Cell viability was evaluated by the MTT assay, gene expression was analyzed by RT-qPCR, and lipid accumulation was determined using Oil Red O staining. Pyrosequencing was performed to measure the methylation of the Pparg promoter region.Results: Aaptamine treatment significantly dose-dependently decreased lipid accumulation and inhibited Pparg and Slc2a4 mRNA expression. However, there were no significant differences in the methylation level of the Pparg promoter.Conclusion: Aaptamine inhibits lipid accumulation and Pparg and Slc2a4 mRNA expression while maintaining the methylation level of the Pparg promoter during 3T3-L1 adipocyte differentiation.Keywords: aaptamine, adipocyte differentiation, DNA methylation, Pparg, Slc2a4