不同浓度牛乳铁蛋白对破骨细胞活性的影响

摘要

目的 研究不同浓度牛乳铁蛋白对于破骨细胞的活性的影响.方法 首先通过噻唑蓝(methyl thiazolyl tetrazolium, MTT)法检测不同浓度的牛乳铁蛋白对RAW264.7的细胞毒性, 进而在核因子κB受体活化因子配体(receptor activator of the NF-κB ligand, RANKL)(100 ng/mL)诱导下形成破骨细胞.通过抗酒石酸酸性磷酸酶(tartrate resistant acid phosphatase, TRAP)染色法观察阳性多核细胞, 检测不同浓度的牛乳铁蛋白对于破骨细胞形成的影响.通过TRAP活性检测试剂盒检测上清中的TRAP活性, 及通过人工骨板观察破骨细胞的骨吸收功能.结果 牛乳铁蛋白对RAW264.7细胞无毒性, 对破骨细胞的形成有显著的抑制作用, 并且能够显著地降低破骨细胞的活性, 对破骨细胞的骨吸收活性也有抑制作用.结论 牛乳铁蛋白对破骨细胞的活性有显著影响, 可能会在以破骨细胞活性过度表达为特征的骨病中起到重要作用.%Objective To investigate the effects of different concentrations of bovine lactoferrin on the activity of osteoclasts. Methods First, thiazolyl tetrazolium (MTT) method was used to detect the cytotoxicity of different concentrations of bovine lactoferrin in RAW264.7, and then osteoclast cells were induced by the receptor activator of the NF-κB ligand (RANKL) (100 ng/mL). The effects of different concentrations of the bovine lactoferrin on the formation of osteoclasts were examined by tartrate resistant acid phosphatase (TRAP) staining. The TRAP activity was detected by TRAP activity test kit, and the bone absorption function of osteoclasts was observed through artificial bone plate. Results Lactoferrin had no toxicity to RAW264.7 cells, and had a significant inhibitory effect on the formation of osteoclast cells. Moreover, the activity of osteoclast could be significantly reduced, and the bone absorption activity of osteoclasts could also be inhibited. Conclusion Bovinelactoferrin has significant effect on osteoclast viability and might play an important role in regulating bone diseases characterized by excessive osteoclast activity.