于哲,王静凤,毛磊,等.鲫鱼卵唾液酸糖蛋白抑制小鼠破骨细胞的分化机制研究[J].中国海洋药物,2015,34(5):33-40. |
鲫鱼卵唾液酸糖蛋白抑制小鼠破骨细胞的分化机制研究 |
The mechanisms of Carassius auratus sialyglycoproteins in suppressing the differentiation of osteoclast |
投稿时间:2015-03-27 修订日期:2015-04-18 |
DOI: |
中文关键词: 鲫鱼卵唾液酸糖蛋白 抗酒石酸酸性磷酸酶 核因子κB 丝裂原活化蛋白激酶 |
English Keywords:Ca-SGP TRAP NF-κB MAPK |
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中文摘要: |
目的:探讨鲫鱼卵唾液酸糖蛋白(Carassius auratus sialyglycoproteins,Ca-SGP)对核因子κB受体活化因子配体(receptor activator of NF-κB ligand,RANKL)诱导形成的破骨细胞的抑制作用及机制。方法:初断乳ICR雄性小鼠,无菌取股骨,分离骨髓造血细胞,通过诱导剂巨噬细胞集落刺激因子(macrophage colony stimulating factor,M-CSF)和RANKL诱导破骨细胞的分化与成熟。MTT法检测Ca-SGP对破骨细胞及其前体细胞活力的影响;通过抗酒石酸酸性磷酸酶(tartrate-resistant acid phosphatase,TRAP)染色、TRAP检测试剂盒和骨吸收陷窝实验测定Ca-SGP在破骨细胞分化与成熟过程中的作用;实时荧光定量PCR法(qRT-PCR)测定破骨细胞形成过程中核因子κB(Nuclear factor,NF-κB)和丝裂原活化蛋白激酶(mitogen-activated protein kinases,MAPK)信号通路关键因子的表达。结论:Ca-SGP可有效抑制由RANKL诱导的破骨细胞分化及成熟,降低细胞TRAP活性及其表达,抑制骨吸收陷窝形成;分子机制方面,Ca-SGP可明显降低信号传导关键蛋白TRAP6的活性,进而抑制NF-κB和MAPKs信号通路关键因子mRNA的表达,达到抑制破骨细胞分化成熟的效果。结果:Ca-SGP可通过NF-κB和MAPKs信号通路抑制由RANKL诱导的破骨细胞形成和分化成熟。 |
English Summary: |
Purpose:To investigate the effects of Ca-SGP on the RANKL-induced osteoclastogenesis and signaling pathways. Method: Osteoclast were differentiation from mouse bone marrow-derived macrophages(BMMs) with M-CSF and RANKL. Osteoclast and its precursor activity were measured by MTT. Osteoclast numbers were evaluated by TRAP stains. mRNA levels were analyzed by qRT-PCR. Results: Ca-SGP inhibited the osteoclast differentiation without toxicity ,and reduced the generation of TRAP-positive multinucleated cells by suppressing MAPK and NF-κB signaling pathways. Conclusion: Ca-SGP suppresses osteoclastogenesis through inhibiting RANKL-induced MAPKs and NF-κB pathway. |
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