天津医药

• 实验研究 •    

破骨细胞抑制性凝集素胞外段可抑制骨髓干细胞向破骨细胞分化

刘瑞1,王宝利2,戴晨琳3,邱明才4,郑纺5,郭善一6   

  1. 1. 天津医大内分泌研究所
    2. 天津医科大学代谢病医院内分泌研究所、卫生部激素与发育重点实验室
    3. 天津医大总医院内分泌科
    4. 天津医科大学总医院
    5. 天津中医药大学
    6. 天津医科大学内分泌研究所
  • 收稿日期:2010-01-25 修回日期:2010-04-15 出版日期:2010-08-15 发布日期:2010-08-15
  • 通讯作者: 戴晨琳

Inhibition of Osteoclast Formation by hOCIL Extracellular Domain

  • Received:2010-01-25 Revised:2010-04-15 Published:2010-08-15 Online:2010-08-15

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摘要： 摘要：目的：破骨细胞抑制性凝集素（Osteoclasst inhibitory lectin，OCIL）是由成骨细胞/基质细胞谱系表达的Ⅱ型跨膜蛋白。在大肠杆菌中表达OCIL胞外段重组蛋白并做功能分析，来研究其对破骨细胞形成的作用。方法：通过修改OCIL胞外段编码序列的碱基序列和重组PCR方法得到连续DNA片段；将表达载体pMAL-c2x/hOCIL转化入BL21(DE3)plysS宿主菌， IPTG诱导表达。结果：可见细胞裂解物上清和沉淀中均有目的蛋白表达；体外培养小鼠股骨骨髓细胞，在M-CSF和RANKL诱导骨髓细胞向破骨细胞分化的基础上，加入不同浓度的重组蛋白，结果显示所制备的重组蛋白对小鼠破骨细胞的形成有抑制作用（p＜0.05）。结论：本研究通过基因工程的方法成功获得了hOCIL重组蛋白，此蛋白在体外可以明显抑制破骨细胞样细胞的形成。

关键词: 破骨细胞抑制凝集素, 表达, 重组PCR, 重组蛋白, 破骨细胞

Abstract: [Abstract] Objectives:Osteoclasst inhibitory lectin (OCIL) expressed by the osteoblast/stomal cells is a type-Ⅱ transmembrane protein. OCIL extracellular domain was obtained to investigate its effect on the formation of osteoclast like cells. Mehtods: based on the codon preference and degeneration, the codons of OCIL extracellular domain was changed into those preferred by E.coli coding and ligated the fragments with recombinant PCR. By induction of IPTG, the recombinant cells BL21(DE3)plysS with transfection of pMAL-c2x/hOCIL produced the soluble recombinant protein in the cell lysates. then the combination activity between maltose binding protein (MBP) and Amylose was used to purify the fusion protein. The biological action of the recombinant protein was analyzed by its effect on osteoclast formation in murine bone marrow cell culture under the stimulation of macrophage-colony stimulating factor (M-CSF) and receptor activator of nucleic factor κB ligand (RANKL). Results: osteoclast like cells (OLCs) formation was significantly blocked by the recombinant protein. Conclusion: the research successfully obtained the recombinant protein hOCIL from E.coli with biological activity that may inhibited the osteoclast like cells formation in vitro.

Key words: Osteoclast inhibitory lectin, expression, recombinant PCR, recombinant protein, Osteoclast