白细胞介素29抑制破骨细胞分化的研究

Inhibitory effect of interleukin 29 on osteoclast differentiation

目的 :新型细胞因子白细胞介素(interleukin,IL)-29具有抗病毒、抗肿瘤、免疫调节等作用,但其在破骨细胞分化中的作用尚未见报道。本研究探讨IL-29对核因子κB受体活化因子配基(receptor activation of nuclear-κB ligand,RANKL)诱导小鼠巨噬细胞RAW264.7(破骨前体细胞)向破骨细胞分化的作用。方法:流式检测IL-29特异性受体IL-28Rα在RAW264.7细胞表面的表达,CCK8法检测IL-29对RAW264.7细胞增殖的影响。不同浓度IL-29重组因子加入到RANKL诱导的RAW264.7向破骨细胞分化的体系中,5 d后通过抗酒石酸酸性磷酸酶染色检测IL-29对破骨细胞数目的影响。实时荧光定量PCR检测IL-29对破骨细胞标志性基因TRAP、CTR、CTSK及相关基因TRAF6、RANK表达的影响。结果 :IL-29呈剂量依赖性抑制RANKL诱导的RAW264.7中破骨细胞的形成,100 ng/m L IL-29可显著抑制破骨细胞的形成以及破骨细胞形成相关基因TRAP、CTR、CTSK、TRAF6、RANK的表达。结论 :本研究首次证实IL-29可以抑制破骨细胞形成及其功能。

Objective：The novel cytokine interleukin（IL）-29 has been reported to induce antiviral, antitumor, and immune responses. However,the effect of IL-29 on osteoclast differentiation remains unclear. This study aimed to explore the effect of IL-29 on osteoclast differentiation and function in RANKL-induced in RAW264.7 cells（osteoclast precursors）. Methods：The expression of IL-28Rα（specific receptor of IL-29） on RAW264.7 cells was evaluated by flow cytometry. RAW264.7 cells were stimulated with RANKL and different doses of recombinant IL-29 for 5 d,then the osteoclast formation was evaluated by counting multinucleated cells for tartrate-resistant acid phosphatase（TRAP） staining. Furthermore,the effect of IL-29 on the RANKL-induced expression of osteoclastic genes（TRAP,CTR and CTSK） and relevant genes（TRAF6 and RANK） was detected by real time PCR. Results：IL-29 inhibited the osteoclast formation in a dose-dependent manner in RANKL-induced RAW264.7 cells,and the dose of 100 ng / ml IL-29 significantly reduced the number of TRAP positive multinucleated cells and m RNA expression of osteoclastic genes（TRAP,CTR and CTSK） and relevant genes（TRAF6 and RANK）. Conclusion：The findings in the present study indicate,for the first time,that IL-29 could inhibit osteoclast formation and function.