Anti-adipogenic effects of gancaonin N, a bioactive compound from Glycyrrhiza uralensis, in MDI-Induced 3T3-L1 adipocytes

Abstract

Obesity represents a significant global health challenge resulting from the intricate interactions between genetic predispositions, environmental influences, and lifestyle choices, ultimately causing abnormal fat accumulation. Glycyrrhiza uralensis , a plant traditionally used in Asian herbal medicine, has anti-inflammatory and antioxidant properties and has been reported to influence metabolic processes. Gancaonin N (GN), a prenylated isoflavone derived from G. uralensis , has not been investigated for its potential effects on adipogenesis and lipid metabolism. This study employed a combined approach using network pharmacology and in vitro validation in 3T3-L1 adipocytes. Network pharmacology was used to predict GN-related targets, identify overlapping genes with obesity, and perform Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses. Subsequently, 3T3-L1 preadipocytes were treated with GN to evaluate its effects on adipocyte differentiation and lipid accumulation. Oil Red O staining, Western blotting, and PCR analyses were conducted to assess adipogenesis and lipid metabolism related pathways. Network pharmacology analysis identified 17 overlapping targets between GN-related and obesity-related genes, with key hub genes including peroxisome proliferator-activated receptor γ (PPARγ) and fatty acid synthase (FASN). GO and KEGG enrichment analyses suggested that GN is associated with pathways related to lipid metabolism, adipogenesis, and AMP-activated protein kinase (AMPK) signaling. To validate these predictions, 3T3-L1 cells were treated with GN. Adipocyte differentiation and lipid accumulation were significantly inhibited in a concentration-dependent manner, as demonstrated by Oil Red O staining. Western blot and PCR analyses revealed downregulation of key adipogenic regulators, including PPARγ, CCAAT/enhancer-binding protein α (C/EBPα), and sterol regulatory element-binding protein 1c (SREBP-1c), as well as suppression of lipogenic genes such as FASN and FABP4. Moreover, GN was associated with activation of AMPK, a central regulator of energy homeostasis and fatty acid oxidation. These findings suggest that GN may regulate adipocyte differentiation and lipid metabolism through multiple signaling pathways.