Huang An, Gu Zhiping, Jin Jiahao, Nie Tao
Orthopedic Hospital, Postdoctoral Innovation Practice Bace, The First Affiliated Hospital, Jiangxi Medical College, Nanchang University, Nanchang 330006, P.R. China.
Orthopedic Hospital, The First Affiliated Hospital, Jiangxi Medical College, Nanchang University, Nanchang, P.R. China.
J Microbiol Biotechnol. 2025 Feb 13;35:e2409009. doi: 10.4014/jmb.2409.09009.
This study investigates the effects of casticin on osteoclastogenesis, aiming to elucidate its underlying mechanisms for potential clinical applications. We assessed the cytotoxicity of casticin using a CCK assay in RAW 264.7cell (murine cell line, from ATCC), which differentiate into osteoclasts upon RANKL treatment. Various concentrations (0.125, 0.25, 0.50 μM) were tested to establish a dose-dependent response. The effects of casticin on osteoclast differentiation and actin filament organization were evaluated through TRAP and F-actin staining. Additionally, qPCR and Western blot analyses were performed to assess gene expression. Concentrations exceeding 1.00 μM caused significant cytotoxicity. Notably, casticin at 0.50 μM significantly inhibited osteoclast differentiation and function, reducing marker gene expression, including c-FOS, NFATc1, CtsK, and MMP-9. Furthermore, casticin decreased phosphorylation levels of NF-κB and IκBα and downregulated BCL-2 expression. Our findings highlight casticin's potent regulatory effects on osteoclasts via the NF-κB/BCL-2 signaling pathways, suggesting potential therapeutic applications in bone-related disorders.
本研究调查了紫花牡荆素对破骨细胞生成的影响，旨在阐明其潜在临床应用的潜在机制。我们使用CCK检测法评估了紫花牡荆素对RAW 264.7细胞（小鼠细胞系，来自美国典型培养物保藏中心）的细胞毒性，该细胞在RANKL处理后可分化为破骨细胞。测试了各种浓度（0.125、0.25、0.50 μM）以建立剂量依赖性反应。通过TRAP和F-肌动蛋白染色评估了紫花牡荆素对破骨细胞分化和肌动蛋白丝组织的影响。此外，进行了qPCR和蛋白质印迹分析以评估基因表达。浓度超过1.00 μM会引起明显的细胞毒性。值得注意的是，0.50 μM的紫花牡荆素显著抑制破骨细胞分化和功能，降低包括c-FOS、NFATc1、CtsK和MMP-9在内的标志物基因表达。此外，紫花牡荆素降低了NF-κB和IκBα的磷酸化水平，并下调了BCL-2表达。我们研究结果突出了紫花牡荆素通过NF-κB/BCL-2信号通路对破骨细胞具有强大的调节作用，提示其在骨相关疾病中的潜在治疗应用。
