Bilobalide exhibits numerous beneficial bioactivities, including neuroprotective, anti-inflammatory, and antioxidant activity. Our previous study demonstrated that bilobalide inhibits adipogenesis and promotes lipolysis. The dose-dependent cytotoxicity was found to be specific to the mature adipocytes only, indicating the potential for regulating apoptosis in them. Herein, we aimed to investigate the apoptotic effects of bilobalide on 3T3-L1 mature adipocytes and elucidate the underlying mechanisms thereof. Flow cytometry analysis (FACS) revealed the pro-apoptotic effects of bilobalide on these cells. Bilobalide induced early apoptosis by reducing the mitochondrial membrane potential (MMP). DNA fragmentation was confirmed using TUNEL staining. Additionally, bilobalide increased the intracellular reactive oxygen species (ROS) levels and activities of Caspases 3/9. Pre-treatment with NAC (an ROS scavenger) confirmed the role of ROS in inducing apoptosis. Moreover, bilobalide up- and down-regulated the expression of Bax and Bcl-2, respectively, at the mRNA and protein expression levels; upregulated the Bax/Bcl-2 ratio; triggered the release of cytochrome c from the mitochondria; and increased the protein expression of cleaved Caspase 3, cleaved Caspase 9, and PARP cleavage. These results support the conclusion that bilobalide induces apoptosis in mature 3T3-L1 adipocytes through the ROS-mediated mitochondrial pathway, and offers potential novel treatment for obesity.

Obesity is a major health crisis that impacts social and economic burdens worldwide, and it is being studied intensively to understand the molecular mechanisms and regulators of this complex condition. Over the past decade, significant insights into adipocyte fate have been gained, demonstrating that adipocyte behavior is influenced by intracellular pathways, systemic mediators, and environmental signals. In this review, we discuss the current understanding of key molecular and functional changes in adipose tissue. We focused on ER–mitochondrial dynamics and their role in beige adipocytes. We examined the biological properties of beige fat cells, including their inflammatory status, redox balance, and regulation of adipocyte transdifferentiation via signaling molecules. Finally, we explored the effects of naturally derived compounds on adipocyte function and their potential to combat obesity using a comprehensive approach. The process of adipocyte beiging induced by coordinated intracellular signaling pathways highlights mechanisms that can reprogram metabolic homeostasis and reduce obesity-related phenotypes.