Bilobalide Induces Apoptosis in 3T3-L1 Mature Adipocytes through ROS-Mediated Mitochondria Pathway

Abstract

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.

Enhancing Physicochemical Properties and Biocompatibility of Hollow Porous Iron Oxide Nanoparticles through Polymer-Based Surface Modifications

“…This could be because MCF10A cells do not possess the same robust mechanisms as RAW264.7 cells for managing elevated iron concentrations, which are characterized by efficient iron storage via ferritin and restricted iron export via ferroportin . In the case of 3T3-L1 cells, their heightened resistance can be attributed to lower levels of proapoptotic proteins, such as Bax and Bak, which diminish their susceptibility to the treatments. , …”

“…54 In the case of 3T3-L1 cells, their heightened resistance can be attributed to lower levels of proapoptotic proteins, such as Bax and Bak, which diminish their susceptibility to the treatments. 55,56 The surface modification with polymers did not induce substantial cellular damage in normal cells. This, combined with the negligible cytotoxicity in uncoated particles, presents a promising prospect, indicating the potential suitability of these coated particles as carriers of biologically active molecules.…”

Safety and efficacy of a feed additive consisting of a dry extract obtained from the leaves of Ginkgo biloba L. (ginkgo extract) for horses, dogs, cats, rabbits and guinea pigs (FEFANA asbl)

“…Oxidative stress was observed after treatment of the cells with bilobalide at 1 and 2 mM (corresponding to 326.3 and 652.5 μg/mL) during 4 h, whereas the ginkgolides reduced the ROS formation at the same concentration and time. ROS induced apoptosis was also observed in 3T3‐L1 mature adipocytes in culture at treatment with bilobalide (Bu et al., 2023 ).…”

Osmanthus fragrans Flavonoid Extract Inhibits Adipogenesis and Induces Beiging in 3T3-L1 Adipocytes

“…Intracellular ROS production in mature adipocytes after OFFE treatment were assessed at various time points using an ROS assay kit (Beyotime, Shanghai, China). The procedure was followed as previously described [ 68 ]. The 3T3-L1 pre-adipocytes were differentiated in 12-well culture plates.…”