促炎细胞因子对体外诱导小鼠破骨样细胞的影响

Influence of proinflammatory cytokines in inducing mouse osteoclast-like cell formation in vitro

目的探索促炎因子白细胞介素(IL)-1、IL-6和肿瘤坏死因子(TNF)-α对体外诱导小鼠破骨样细胞的影响。方法从小鼠脾脏中提取出破骨细胞前体细胞,用不同细胞因子分组诱导:A组为巨噬细胞集落刺激因子(M-CSF);B组为M-CSF和溶解型核因子-κB受体活化因子配体(sRANKL);C组为M-CSF、SRANKL、IL-1、IL-6、TNF-α;D组为M-CSF、IL-1、IL-6、TNF-α。用倒置显微镜和抗酒石酸酸性磷酸酶(TRAP)染色观察细胞分化情况,使用扫描电子显微镜观察骨磨片。结果 C、B组破骨样细胞数目均明显多于A、D组(P<0.05),且C组高于B组(P<0.05);D、A组没有诱导出破骨样细胞(P>0.05)。结论促炎因子TNF-α、IL-1、IL-6可通过sRANKL在体外促进小鼠破骨样细胞的分化,单独的TNF-α、IL-1、IL-6不能够诱导出破骨样细胞。

Objective This study aims to investigate the influence of proinflammatory cytokines interleukin（IL）-1, IL-6, and tumor necrosis factor（TNF）-α on differentiation and formation of mouse osteoclast-like cells in vitro. Methods Osteoclast precursor cells from mice spleen were cultured under four conditions. Group A was cultured with macrophage colony stimulating factor（M-CSF）. Group B was cultured with M-CSF and the soluble ligand of receptor activator of NF- κB（sRANKL）. Group C was cultured with M-CSF, sRANKL, IL-1, IL-6, and TNF-α. Finally, Group D was cultured with M-CSF, IL-1, IL-6, and TNF-α. All groups were cultured in 24-well plates with bone slices. Tartrate-resistant acid phosphatase staining and inverted microscope were employed to determine differentiation. A scanning electron microscope was used to examine the bone slices. Results The number of osteoclast-like cells from Groups C and B was significantly higher than that in Groups A and D（P〈0.05）. The number of cells in Group C was greater than that in Group B（P〈0.05）. No osteoclast-like cells were found in Groups A and D（P〉0.05）. Conclusion Proinflammatory cytokines IL-1, IL-6, and TNF-α can promote the differentiation of osteoclast-like cells in vitro, which is dependent on sRANKL. These cytokines cannot directly induce the formation of osteoclast-like cells.