College of Biology and the Environment, Nanjing Forestry University, Nanjing 210037, China.
College of Forestry, Nanjing Forestry University, Nanjing 210037, China.
Molecules. 2023 Sep 2;28(17):6410. doi: 10.3390/molecules28176410.
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.
银杏叶内酯具有多种有益的生物活性，包括神经保护、抗炎和抗氧化活性。我们之前的研究表明，银杏叶内酯抑制脂肪生成并促进脂肪分解。发现剂量依赖性细胞毒性仅对成熟脂肪细胞具有特异性，表明其有可能调节脂肪细胞的细胞凋亡。在此，我们旨在研究银杏叶内酯对 3T3-L1 成熟脂肪细胞的细胞凋亡作用，并阐明其潜在机制。流式细胞术分析（FACS）显示银杏叶内酯对这些细胞具有促凋亡作用。银杏叶内酯通过降低线粒体膜电位（MMP）诱导早期细胞凋亡。TUNEL 染色证实了 DNA 片段化。此外，银杏叶内酯增加了细胞内活性氧（ROS）水平和 Caspase 3/9 的活性。用 NAC（ROS 清除剂）预处理证实了 ROS 在诱导细胞凋亡中的作用。此外，银杏叶内酯分别上调和下调了 Bax 和 Bcl-2 的 mRNA 和蛋白表达水平；上调了 Bax/Bcl-2 比值；触发了细胞色素 c 从线粒体中的释放；并增加了裂解的 Caspase 3、裂解的 Caspase 9 和 PARP 裂解的蛋白表达。这些结果支持以下结论：银杏叶内酯通过 ROS 介导的线粒体途径诱导成熟 3T3-L1 脂肪细胞凋亡，并为肥胖症提供了潜在的新治疗方法。
