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| 骨保护素体外抑制小鼠单核巨噬细胞 成熟分化的实验研究 |
| Study on osteoprotegerin in inhibitation of mouse osteoclast precursor cell differentiation |
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| DOI： |
| 中文关键词: 破骨细胞；骨质疏松；RAW264.7细胞；HANKL 0PG |
| 英文关键词:Osteoclasts Osteoporosis RAW264.7 cells RANKL OPG |
| 基金项目: |
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| 中文摘要: |
| 目的研究骨保护素（Osteoprotegerin, 0PG)抑制核因子NF-KB受体活化因子配体（Receptor activator of nuclear kappa B ligand，RANKL)诱导小鼠单核细胞RAW264. 7成熟分化而导致的溶骨效 应。方法50 ng/mL RANKL诱导RAW264. 7细胞1 d后，加人100 ng/mL 0PG(实验组，即0PG + RANKL组）或不加人0PG(对照组，即RANKL组）分别培养7 d和9 d，经细胞形态学观察其变化，抗 酒石酸酸性碟酸酶（Tartrate resistant acid phosphatase, TRAP)染色法观察TRAP阳性多核细胞，扫描 电镜下观察在骨片上的破骨细胞所致的骨吸收陷窝形成情况。结果对照组培养7 d时，在倒置相 差显微镜、透射电镜、光镜下可见细胞形状为椭圆形或不规则形，胞体明显较KAW264.7细胞增大， 胞核多为6 ~ 10个，扫描电镜下还可见大量伪足形成，而实验组培养7 d后，细胞形状多为圆形，且扫 描电镜下未见明显伪足形成；对照组9 d时可见大量TRAP染色阳性的多核巨细胞（含3个或3个以 上的细胞核），而实验组中TRAP染色阳性的多核破骨细胞偶见多核巨细胞，培养9 d时很难找到多 核巨细胞；仅用RANKL诱导RAW264.7细胞分化7 d时，对照组中破骨细胞表面可见大量伪足伸出， 并形成明显的骨吸收陷窝，实验组中破骨细胞见少许伪足突出，不能看到明显的骨陷窝形成。结论 单用50 ng/mL RANKL体外连续诱导RAWM4.7细胞7 d时，可以促进成熟的破骨细胞显著分化。 100 ng/mL 0PG培养9 d能有效地抑制破骨细胞的分化，减少破骨细胞的骨吸收效应。 |
| 英文摘要: |
| Objective To observe whether OPG is effective in inhibiting the effect that RANKL induce RAW264. 7 cells to differentiation to mature osteoclast. Methods RAW264. 7 cells were solely treated with 50 ng/ml RANKL for 1 day, which then were divided into two groups： the one is an OPG group involving 100 ng/ml OPG and 50 ng/ml RANKL, the other is a control group only containing 50 ng/ml RANKL. After the period between 7 and 9 d, cells morphological changes can be investigated by Inverted Phase Control Microscope (IPCM )，Transmission Electron Microscope ( TEM )，Scanning Electron Microscope ( SEM ) and Light Microscope respectively ； Furthermore, Staining THAP-positive multinucleated cells can be detected by IPCM;The resorption pits of bone slices were indicated through SEM. Results It is very clear that the sharp of RAW264.7 cells became oval or irregular, and their bodies go bigger significantly including 6 up to 10 nucleus in control group by means of IPCM, TEM and Light Microscope. Interestingly, we also have found that there were considerable pseudopodia-like protrusions attaching to the surface of these cells by Scanning Electron Microscopy in the control group. By contrast, the research result is polar in OPG group. |
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