Hu Ke-Fei, Kong Xiang-Ying, Zhong Mi-Cun, Wan Hong-Ye, Lin Na, Pei Xiao-Hua
Beijing University of Chinese Medicine, Beijing, 100029, China.
Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing, 100700, China.
Chin J Integr Med. 2017 Feb;23(2):110-116. doi: 10.1007/s11655-016-2647-2. Epub 2016 Dec 29.
To examine the effects of brucine on the invasion, migration and bone resorption of receptor activator of nuclear factor-kappa B ligand (RANKL)-induced osteoclastogenesis.
The osteoclastogenesis model was builded by co-culturing human breast tumor MDA-MB-231 and mouse RAW264.7 macrophages cells. RANKL (50 ng/mL) and macrophage-colony stimulating factor (50 ng/mL) were added to this system, followed by treatment with brucine (0.02, 0.04 and 0.08 mmol/L), or 10 μmol/L zoledronic acid as positive control. The migration and bone resorption were measured by transwell assay and in vitro bone resorption assay. The protein expressions of Jagged1 and Notch1 were investigated by Western blot. The expressions of transforming growth factor-β1 (TGF-β1), nuclear factor-kappa B (NF-κB) and Hes1 were determined by enzyme-linked immunosorbent assay.
Compared with the model group, brucine led to a dose-dependent decrease on migration of MDA-MB-231 cells, inhibited RANKL-induced osteoclastogenesis and bone resorption of RAW264.7 cells (P<0.01). Furthermore, brucine decreased the protein levels of Jagged1 and Notch1 in MDA-MB-231 cells and RAW264.7 cells co-cultured system as well as the expressions of TGF-β1, NF-κB and Hes1 (P<0.05 or P<0.01).
Brucine may inhibit osteoclastogenesis by suppressing Jagged1/Notch1 signaling pathways.
研究马钱子碱对核因子κB受体激活剂配体（RANKL）诱导的破骨细胞生成、侵袭、迁移及骨吸收的影响。
通过共培养人乳腺癌MDA-MB-231细胞和小鼠RAW264.7巨噬细胞构建破骨细胞生成模型。向该体系中加入RANKL（50 ng/mL）和巨噬细胞集落刺激因子（50 ng/mL），随后用马钱子碱（0.02、0.04和0.08 mmol/L）处理，或用10 μmol/L唑来膦酸作为阳性对照。采用Transwell实验和体外骨吸收实验检测迁移和骨吸收情况。通过蛋白质免疫印迹法研究Jagged1和Notch1的蛋白表达。采用酶联免疫吸附测定法测定转化生长因子-β1（TGF-β1）、核因子κB（NF-κB）和Hes1的表达。
与模型组相比，马钱子碱使MDA-MB-231细胞的迁移呈剂量依赖性降低，抑制RANKL诱导的RAW264.7细胞破骨细胞生成和骨吸收（P<0.01）。此外，马钱子碱降低了MDA-MB-231细胞与RAW264.7细胞共培养体系中Jagged1和Notch1的蛋白水平以及TGF-β1、NF-κB和Hes1的表达（P<0.05或P<0.01）。
马钱子碱可能通过抑制Jagged1/Notch1信号通路抑制破骨细胞生成。
