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目的:研究健脾活血补肾汤对骨髓源性破骨细胞形成及其骨吸收功能的影响。方法:55只SD雄性大鼠,随机分成5组,每组11只,除正常组外其余4组均给予弗式完全佐剂(FCA)建立佐剂型关节炎(AA)模型。造模第3天起高、中、低剂量组大鼠分别给予高、中、低浓度健脾活血补肾汤,正常组、模型组大鼠予以等量的0.9%氯化钠溶液。灌胃30d后,获取各组大鼠骨髓巨噬细胞(BMM)。接种入含或不含牛骨片的24孔板中,利用核转录因子κβ受体活化因子配体和巨噬细胞集落刺激因子共同诱导小鼠骨髓巨噬细胞分化成破骨细胞。苏木精-伊红染色法及抗酒石酸酸性磷酸酶染色进行破骨细胞分化鉴定并计数,计算机图像分析技术测定骨片上骨吸收陷窝的面积。结果:健脾活血补肾汤各组破骨细胞数量及陷窝吸收面积均低于模型组(P<0.05),高剂量组破骨细胞数量低于中、低剂量组(P<0.05)。结论:健脾活血补肾汤可以抑制AA大鼠体外细胞因子诱导的破骨细胞的形成及其骨吸收功能。
Objective: To study the effect of Jianpi Huoxue Bushen Decoction on bone marrow-derived osteoclast formation and its bone resorption function. Methods: Fifty-five male Sprague Dawley rats were randomly divided into 5 groups (11 rats in each group). Adjuvant arthritis (AA) model was established by complete Freund’s adjuvant (FCA). Rats in high, medium and low dose groups were given high, medium and low concentration of Jianpi Huoxue Bushen Decoction on the third day after model making. The rats in normal group and model group were given the same amount of 0.9% sodium chloride solution. After intragastric administration for 30 days, the bone marrow macrophages (BMMs) of rats in each group were obtained. Inoculated into a 24-well plate with or without bovine bone chips, the use of nuclear factor kappa β receptor activating factor ligand and macrophage colony stimulating factor co-induced murine bone marrow macrophage differentiation into osteoclasts. Hematoxylin-eosin staining and tartrate-resistant acid phosphatase staining were used to identify and count osteoclast differentiation. The area of bone resorption lacuna in the bone chips was determined by computer image analysis. Results: The numbers of osteoclasts and the area of lacunae in each group were lower than those in model group (P <0.05). The number of osteoclasts in high dose group was lower than that in middle and low dose groups (P <0.05). Conclusion: Jianpi Huoxue Bushen Decoction can inhibit cytokine-induced osteoclast formation and bone resorption in AA rats.