鲫鱼卵唾液酸糖蛋白对小鼠破骨细胞活性的影响

EFFECTS OF CARASSIUS AURATUS SIALYGLYCOPROTEINS ON OSTEOCLAST ACTIVITY IN MICE

目的研究鲫鱼卵唾液酸糖蛋白(Carassius auratus sialyglycoproteins,CA-SGP)对破骨细胞活性的影响。方法通过M-CSF和RANKL体外诱导小鼠骨髓造血细胞分化,建立破骨细胞模型,以MTT法检测CA-SGP对破骨细胞前体细胞及破骨细胞增殖活性的影响;采用TRAP染色和TRAP含量测定法评价其对小鼠骨髓细胞向破骨细胞分化的影响;利用酶联免疫吸附法(enzyme-linked immunosorbent assay,ELISA)检测CA-SGP对小鼠破骨细胞促炎症因子IL-6、IL-1β和TNF-α分泌的影响。结果 CA-SGP对破骨细胞前体细胞和破骨细胞的增殖活性无影响,可明显抑制小鼠骨髓造血细胞向破骨细胞的分化,且在早期抑制作用最强。ELISA试剂盒检测结果表明,CA-SGP能明显降低小鼠破骨细胞IL-6、IL-1β和TNF-α的含量。结论鲫鱼卵唾液酸糖蛋白可明显抑制小鼠骨髓造血细胞向破骨细胞的分化以及破骨细胞炎症因子的分泌。

Objective To investigate the effects of Carassius auratus sialyglycoproteins（CA-SGP） on osteoclasts activity in mice. Methods The murine osteoclast model was generated by hemopoietic cell differentiation induced by receptor activator for nuclear factor-кB ligand（RANKL） and macrophage colony-stimulating factor（M-CSF） in vitro. The cells were divided into four groups： the normal control group, the model control group and two CA-SGP groups with molecular weight more than or less than 10 ku. MTT assay was used to determine the proliferation ability of osteoclast precursor cells and osteoclasts. Besides, the effects of CA-SGP on the differentiation from BMMs to osteoclast cells were carried out by tartrate resistant acid phosphatase（TRAP） staining and TRAP content assay. The contents of IL-6, IL-1β and TNF-α were measured by enzyme-linked immunosorbent assay（ELISA）. Results CA-SGP did not show any effect on proliferation of osteoclast precursor cells and osteoclasts, but could significantly inhibit differentiation from BMMs to osteoclasts, especially at the early period of differentiation. Based on ELISA assay, CA-SGP could remarkably decrease the contents of IL-1β,IL-6 and TNF-α in osteoclasts. Conclusion CA-SGP could inhibit osteoclastogenesis and the secretion of IL-1β, IL-6 and TNF-α.