摘要
目的比较基质金属蛋白酶9(matrix metalloproteinase 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1 d龄SD大鼠四肢长骨获得成熟OC;利用RANKL(100 ng/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12 d组的积分光密度和平均光密度均明显高于机械法3 d组.结论诱导12 d的OLC MMP-9及其mRNA表达的明显高于机械法3 d,为OLC提供了进一步的实验数据.
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.
出处
《昆明医科大学学报》
CAS
2013年第12期1-4,25,共5页
Journal of Kunming Medical University
基金
国家自然科学基金资助项目(30660212
81160401
81260493)
云南省社会发展计划重点项目(2008CC009)
云南省应用基础研究基金资助项目(2009CD214
2004C0044M)
关键词
破骨细胞
破骨样细胞
机械法
诱导法
基质金属蛋白酶9
Osteoclast
Osteoclast like cells
Mechanical method
Inducement method
Matrix metalloproteinase 9
