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目的探讨白三烯(LT)B4能否不依赖细胞核因子资B受体激活剂配体(RANKL)直接促进人破骨细胞的分化和激活。方法阳性对照组用25ng/ml巨噬细胞集落刺激因子(M-CSF)和30ng/mlsRANKL来诱导人外周血单核细胞的培养,实验组用25ng/mlM-CSF和10-9、10-8、10-7mol/LLTB4来诱导。通过抗酒石酸磷酸酶(TRAP)染色及10mm×10mm玻片上多核性TRAP染色(+)的破骨细胞样细胞计数,来确定LTB4的直接分化作用,并与RANKL的作用比较。通过甲苯胺蓝染色10mm×10mm牛皮质骨片上的骨质吸收陷窝并计数,来确定LTB4直接功能激活作用,并与RANKL的作用比较。结果当M-CSF存在时,LTB4能够直接分化人外周血单核细胞为破骨细胞样细胞,并能激活其骨质吸收功能。且随LTB4浓度的增加而增强,但要弱于RANKL。结论LTB4对人破骨细胞有不依赖RANKL的直接分化和激活作用。
Objective To investigate whether leukotriene (LT) B4 can directly promote human osteoclast differentiation and activation independent of RANKL. Methods Positive control group were cultured with 25ng / ml macrophage colony stimulating factor (M-CSF) and 30ng / mlsRANKL to induce the culture of human peripheral blood mononuclear cells. The experimental group was treated with 25ng / ml M-CSF and 10-9,10-8 , 10-7mol / LLTB4 to induce. The direct differentiation of LTB4 was determined by tartaric acid phosphatase (TRAP) staining and osteoclast-like cell counting of multinucleated TRAP staining (+) on a 10 mm x 10 mm slide and compared to the effect of RANKL. The direct functional activation of LTB4 was determined by toluidine blue staining of the bone resorption lacuna on 10 mm x 10 mm bovine cortical bone slices and compared with the effect of RANKL. Results When M-CSF was present, LTB4 could directly differentiate human peripheral blood mononuclear cells into osteoclast-like cells and activate its bone resorption function. And increased with the concentration of LTB4, but weaker than RANKL. Conclusion LTB4 can directly differentiate and activate human osteoclasts without RANKL.