补肾祛湿活血中药对模拟体外骨重塑作用的细胞特征的影响

Effect of Kidney-tonifying, Dampness-dispelling and Blood-activating Chinese Medicine on Bone Remodeling of Cell Model in Vitro

【目的】探讨补肾活血中药(珍骨丸)与祛湿活血中药(骨刺风湿丸)对体外骨重塑作用的影响。【方法】将MC3T3-E1细胞、RAW264.7细胞分别诱导为成骨细胞(osteoblasts,OB)与破骨细胞(osteoclasts,OC),并将两者共培养。通过免疫细胞化学染色鉴定该共培养体系构建成功与否;以灌胃的方法分别将双醋瑞因、珍骨丸及骨刺风湿丸制备成含药血清;运用含药血清处理共培养细胞并进行Western blotting和Real-time PCR分析。【结果】珍骨丸与骨刺风湿丸处理共培养细胞后骨保护素(osteoprotegerin,OPG)表达显著上升(P<0.05),糖蛋白130(glycoprotein 130,gp 130)表达显著下降(P<0.05)。【结论】珍骨丸、骨刺风湿丸通过抑制gp130来抑制OC的骨吸收,并通过RANKL-RANK-OPG系统促进骨形成,增加骨密度达到治疗骨关节炎的作用。

Objective To explore the effect of Zhengu Pills （ a kidney-tonifying and blood-activating Chinese medicine） and Guci Fengshi Pills （ a dampness-dispelling and blood-activating Chinese medicine） on bone remodeling of cell model in vitro. Methods MC3T3-E1 and RAW264.7 cells were differentiated into osteoblasts （ OB） and osteoclasts （ OC） after stimulated by different inductive agents respectively, and then OB and OC were co-cultured. Immunocytochemical staining was applied to identify the construction of co-cultured OB and OC system. The serum containing corresponding medicine was prepared after the rats were given intragastric administration of Diacerein, Zhengu Pills and Guci Fengshi Pills. And then the obtained serum was used for treatment of co-cultured cells separately. At the end of experiment, Western blotting and real-time polymerase chain reaction （PCR） tests were carried out. Results The osteoprotegerin （OPG） expression was increased and gene expression of glycoprotein 130 （gp130） was decreased in co-cultured cells after treatment with the＆amp;nbsp;serum containing Zhengu Pills and Guci Fengshi Pills （ P〈0.05） . Conclusion Zhengu Pills and Guci Fengshi Pills have therapeutic effects on osteoarthritis （ OA） by inhibiting the bone resorption function of OC by the down-regulation of gp130 and by enhancing bone formation and increasing the bone density through recombinant murine’s RANK ligand （RANKL） -RANK-OPG system.