Heat-treated culture-dried Lactiplantibacillus Plantarum L-14 suppresses adipogenesis and lipogenesis in 3T3-L1 and HepG2 cells

Obesity is a growing global health issue prompting the development of probiotic-based interventions. This study investigated whether heat-treated culture-dried L14 (HT-L14) retains these benefits. In HepG2 cells, HT-L14 reduced palmitate-induced lipid accumulation suppressed lipogenic genes including sterol regulatory element-binding protein 1 and carbohydrate-responsive element-binding protein, and activated AMP-activated protein kinase (AMPK) and acetyl-CoA carboxylase (ACC), promoting lipid oxidation. In 3T3-L1 cells, HT-L14 inhibited adipocyte differentiation, reduced the expression of peroxisome proliferator-activated receptor gamma (PPARγ) and CCAAT/enhancer-binding protein alpha (C/EBPα), and increased energy metabolism-related gene expression. RNA sequencing analysis revealed differential gene expression enriched in lipid metabolism and insulin signaling pathways. HT-L14 exhibited no cytotoxicity and demonstrated improved safety compared to live L14 in HepG2 and 3T3-L1 cells. These findings suggest that HT-L14 retains anti-obesity efficacy with enhanced stability, supporting its potential as a therapeutic candidate for obesity and metabolic disorders. • HT-L14 preserved the anti-obesity effects of live L14 with improved stability confirmed in HepG2 and 3 T3-L1 cells. • HT-L14 reduced lipid accumulation and lipogenesis gene expression in HepG2 cells. • HT-L14 inhibited adipogenesis and enhanced energy metabolism in 3 T3-L1 adipocytes. • HT-L14 showed increased EPS yield, suggesting EPS as a key heat-stable factor in its anti-obesity effect. • RNA-seq revealed HT-L14 modulates genes in lipid metabolism, insulin signaling, and AMPK pathways, implying metabolic regulation.