Metabolomic profiling reveals dynamic lipid reprogramming during adipogenesis in 3T3-L1 cells

Objective Adipogenesis, the process of adipocyte differentiation, plays a central role in obesity development. However, the molecular mechanisms underlying adipogenesis and its regulation remain incompletely understood. This study aimed to investigate metabolomic changes during adipocyte differentiation in 3T3-L1 cells and elucidate the associated molecular mechanisms, with potential implications for obesity-related metabolic diseases. Methods Murine 3T3-L1 pre-adipocytes were cultured and induced to differentiate into mature adipocytes over an 8-day period. Samples were collected at pre-differentiation (Pre), 4-day differentiation (Middle) and 8-day differentiation (Mature) stages. Metabolites were extracted using hydrophilic and hydrophobic methods and analyzed by ultra-performance liquid chromatography coupled with mass spectrometry. Data analysis employed kyoto encyclopedia of genes and genomes (KEGG) pathway enrichment analysis to identify differential metabolites and their associated metabolic pathways. Results Significant differences in metabolite profiles were observed among Pre, Middle and Mature. We identified 170 differential metabolites between Pre and Middle (164 increased, 6 decreased), 246 differential metabolites between Pre and Mature (223 increased, 23 decreased), and 124 differential metabolites between Middle and Mature (102 increased, 22 decreased). The most pronounced changes occurred in lipid metabolites, particularly triglycerides and phosphatidylcholines. KEGG enrichment analysis revealed that these differential metabolites were mainly involved in glycerolipid metabolism, glycerophospholipid metabolism, and insulin resistance pathways. Correlation network analysis further identified key genes associated with these metabolites, highlighting the interplay between lipid and amino acid metabolism during adipogenesis. Conclusion This study provides a comprehensive metabolomic profile of adipocyte differentiation in 3T3-L1 cells, revealing significant alterations in lipid metabolism and key metabolic pathways. These findings enhance our understanding of the molecular mechanisms underlying adipogenesis and may contribute to the development of novel diagnostic tools and therapeutic strategies for obesity-related metabolic diseases.