MMP-9表达在不同方法获得破骨细胞的比较研究

目的比较基质金属蛋白酶9（matrix metailoproteinase 9，MMP-9）表达在机械法获得破骨细胞（osteoclasts，OC）和诱导法获得破骨样细胞（osteoclast like cells，OLC）的差异，为体外培养OC提供实验依据．方法采用机械分离方法，从1d龄sD大鼠四肢长骨获得成熟OC；利用RANKL（100ng／mL）和M—CSF（100ng／mL）诱导RAW264．7细胞形成OLC．采用免疫细胞化学和原位杂交技术检测不同方法获得的OC和OLC MMP-9蛋白及其mRNA的表达．结果诱导法12d组的积分光密度和平均光密度均明显高于机械法3d组．结论诱导12d的OLC MMP-9及其mRNA表达的明显高于机械法3d，为OLC提供了进一步的实验数据．

Comparison of MMP-9 Expression in Osteoclasts Obtained by Different Methods

Objective To investigate the differences of protein and mRNA expression of matrix metalloproteinase 9 （MMP-9） in osteoclasts （OC） and osteoclast-like cells （OLC） obtained by mechanical and inducement methods. Methods Mechanical separation method was used to separate mature osteoclasts from long bones of SD rats aged one-day; and inducement culture method was applied to induce OLC by using RANKL （100 ng/mL） and M-CSF （100 ng/mL） . The protein expression of MMP-9 was measured by immunocytochemistry and mRNA expression of MMP-9 was assayed by in situ hybridization. Results The integral optical density （IOD） and average optical density （AOD） of positive cells in the visual field were higher in the 12-d group of inducement method as compared with the 3 d-group of mechanical method. Conclusions It is suggested that the protein and mRNA expression of MMP-9 in OLC obtained by 12 d inducement method is much high than in OC obtained by 3 d mechanical method. OLC obtained by inducement method can be applied in the study of osteoporosis.