氨基多糖对RANKL诱导的RAW264.7向破骨细胞样细胞分化的抑制作用观察

Observation of GAGs inhibiting the osteoclastogenesis of RAW264.7 in presence of RANKL

目的探讨氨基多糖(GAGs)对核因子κB受体活化因子配体(RANKL)诱导的RAW264.7向破骨细胞分化的影响。方法采用RANKL单独诱导RAW264.7细胞(对照组),分别采用不同浓度肝素、高分子量透明质酸(HA)、硫酸软骨素(CS)C联合RANKL诱导RAW264.7分化(分别为肝素组、HA组、CSC组)。细胞培养第3、5、7、8天采用抗酒石酸酸性磷酸酶染色法(TRAP)检测破骨细胞样细胞并计数;ELISA法检测第8天各组培养液上清中抗酒石酸酸性磷酸酶5b(TRAP5b)含量。结果与对照组比较,诱导第3、5、7、8天,破骨样细胞数均明显减少(P<0.05);其中肝素组随浓度的增大而减少(P<0.05)。第8天培养液中TRAP5b含量与细胞计数结果相符。结论氨基多糖对RANKL诱导下RAW264.7向破骨细胞样细胞的分化具有抑制作用;肝素的作用具有剂量依赖性。

Objective To investigate the effects of glycosaminoglycan （GAGs） on the osteoclastogenesis of macro- phage cell line RAW264.7 in presence of receptor activator of nuclear factor-kappa B ligand （RANKL）. Methods RAW264.7 was induced by RANKL alone in control group. Different concentration gradients of heparin, hyaluronic acid （HA） and chondroitin sulfate C（CS） combined with RANKL were used in heparin, HA and CS group. Tartrate resistant acid phosphatase（TRAP） staining was used to evaluate the numbers of osteoclast-like cells in 3,5,7,8 days. The quantities of TRAPSb in the medium were detected by ELISA. Results The osteoclast-like cell numbers in the experimental groups decreased compared to the control groups in 3, 5, 7, 8 days （P 〈 0.05）. The osteoclast-like cell numbers decreased with the increasing of concentration gradients of heparin （P 〈 0. 05 ）. TRAPSb quantities in medium were the same to the count results in day 8. Conclusion GAGs can inhibit osteoclastogenesis of RAW264.7 cell line in presence of RANKL. The in- hibition of heparin is in dose-dependent manner.