破骨细胞抑制因子生物活性定量测定方法的建立  

Development of A Method for Quantitative Determination of Biological Activity of Osteoclast Suppressor

作　　者：杨子义 张晴妮 

机构地区：[1]杰华生物技术(北京)有限公司,北京100102

出　　处：《中国生物制品学杂志》2009年第10期1026-1028,1035,共4页Chinese Journal of Biologicals

基　　金：国家高技术研究发展计划(863计划)资助项目(2002AA214081)

摘　　要：目的以破骨细胞为模型,建立抑制破骨细胞功能药物的生物活性定量测定方法。方法将小鼠RAW264.7和SP2/0细胞在含地塞米松和1,25(OH)2VitD3的破骨细胞诱导分化液中混合培养,通过TRAP染色鉴定破骨细胞的成熟。加入不同浓度的rhOPG-Fc,检测其对RAW264.7细胞增殖、分化和成熟破骨细胞活性的抑制作用,计算药物的IC50,并定量计算活性单位。结果rhOPG-Fc对RAW264.7细胞增殖抑制的IC50为0.02mg/L,其生物活性为50U/mg;对RAW264.7细胞分化抑制的IC50为0.02mg/L,其生物活性为50U/mg;对成熟破骨细胞活性抑制的IC50为0.18mg/L,其生物活性为5.5U/mg。结论以破骨细胞抑制因子为候选药物所建立的细胞学模型,可方便快速地定量测定破骨细胞抑制类药物的生物活性,且能协助判断药物的作用机理。Objective To develop a method for quantitative determination of biological activity of osteoclast suppressor using osteoclast as a model.Methods Murine RAW264.7 and SP 2 /0 cells were co-cultured in DMEM containing dexamethasone and 1,25（OH）2VitD3,and the mature of osteoclasts was identified by TRAP staining.The rhOPG-Fc at various concentrations were added into the RAW264.7 cells,and its inhibiting effect on the proliferation and differentiation of RAW264.7 cells as well as activity of mature osteoclasts were determined,based on which the IC50 and activity unit were calculated.Results The IC50 and biological activity of rhOPG-Fc were 0.02 mg/L and 50 U /mg for inhibiting the proliferation of RAW264.7 cells,0.02 mg /L and 50 U /mg for inhibiting the differentiation of RAW264.7 cells,and 0.18 mg/L and 5.5 U/mg for inhibiting the activity of mature osteoclasts,respectively.Conclusion The cytological model established using OPG as a candidate drug was simple and rapid for determination of biological activity of osteoclast suppressor and was helpful to analysis of mechanism of drugs.

关 键 词：破骨细胞 抑制因子 生物活性 定量测定 

分 类 号：Q-33[生物学]