小檗碱对大鼠骨髓源性破骨细胞的分化及骨吸收功能的影响(英文)

Effects of berberine on differentiation and bone resorption of osteoclasts derived from rat bone marrow cells

目的:研究小檗碱对破骨细胞分化及骨吸收功能的影响,探讨小檗碱抑制骨吸收的细胞学基础。方法:采用原代培养的成骨细胞和骨髓单核细胞联合培养的方法,在1,25-(OH)2维生素D3和地塞米松作用下,使骨髓单核细胞分化形成破骨细胞。通过相差显微镜观察细胞形态,通过抗酒石酸酸性磷酸酶(tartrate-resistant acid phosphatase,TRAP)染色和观察骨片上骨吸收陷窝的形成鉴定破骨细胞。磷酸苯二钠法测定破骨细胞抗酒石酸酸性磷酸酶的活性,计算机图像分析技术测定骨片上破骨性骨吸收陷窝的面积。结果:小檗碱在0.1～10μmol/L范围内,浓度依赖性地抑制TRAP阳性多核破骨细胞的形成和TRAP活性,减少破骨性骨吸收陷窝的面积;在10μmol/L浓度下,对破骨细胞的抑制作用最强,对TRAP阳性多核破骨细胞的形成和TRAP活性的抑制率分别达到了60.45%和42.12%,骨吸收陷窝面积减少72.69%。结论:小檗碱通过抑制破骨细胞的形成、分化和骨吸收功能减少骨质的丢失。

Objective： To observe the effects of berberine on vitro, and to investigate the cellular mechanism of osteoclastic differentiation and bone resorption action in its inhibitory effects on bone resorption. Methods： The multinucleated osteoclasts （MNCs） dexamethasone from bone marrow cells in the coculture were derived by 1, 25-dihydroxyvitamin D3 and system with primary osteoblastic cells. The tartrateresistant acid phosphatase （TRAP） staining and image analysis of bone resorption pit on dental slices were used to identify osteoclast. The activity of TRAP was measured by p-nitrophenyl sodium phosphate assay. The bone resorption pit area on the bone slices formed by osteoclasts was measured by computer image processing. Results： At the concentrations of 0. 1, 1 and 10 μmol/L, berberine dose-dependently suppressed the formation of TRAP-positive multinucleated cells, the TRAP activity and the osteoclastic bone resorption. The strongest inhibitory effect was exhibited at the concentration of 10 μmol/L, with the inhibiting rate of 60.45%, 42.12% and 72.69% respectively. Conclusion： Berberine can decrease bone loss through inhibition of osteoclast formation, differentiation and bone resorption.