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| 模拟微重力条件下重组骨保护素融合蛋白(rhOPG-HSA)对破骨前体细胞Raw264.7抑制效应的研究 |
| Inhibitory effects of rh0PG-HSA on osteoclast precursor Raw264.7 cells under simulated microgravity |
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| DOI： |
| 中文关键词: 模拟微重力 重组骨保护素融合蛋白 破骨前体细胞 抑制效应 |
| 英文关键词:Simulated microgravity rhOPG-HSA Osteoclast precursor cells Inhibitory effect |
| 基金项目:总装后勤部课题资助项目，总后军队十二五重大项目，总后军队十二五面上项目 |
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| 中文摘要: |
| 目的 探讨在模拟微重力(Simulated microgravity，SMG)条件下重组骨保护素融合蛋白(rhOPG-HSA)对破骨前体细胞Raw264.7的抑制效应。方法 在SMG条件下，分别培养破骨前体细胞Raw264.7与成骨细胞MC3T3-E1，利用ELISA法检测MC3T3-E1细胞培养液中骨保护素活性，用巨噬细胞集落刺激因子(M-CSF)以及可溶性破骨细胞分化因子(sRANKL)诱导Raw264.7分化为破骨细胞，通过抗酒石酸酸性磷酸酶(TRAP)染色法鉴定rhOPG-HSA抑制破骨细胞能力，利用半定量RTPCR测定破骨细胞中TRAP的表达。结果 SMG条件下，骨保护素活性在72 h后显著降低(P<0.01)；Raw264.7细胞经M-CSF及sRANKL诱导3、4、5 d后，与阴性对照组相比，TRAP阳性染色的破骨细胞用rhOPG．HSA处理后明显减少(P<0.05)，RT-PCR测定结果表明破骨细胞TRAP的表达降低(P<0.05)。结论 SMG条件下，rhOPG-HSA能够抑制破骨前体细胞Raw264.7的分化。 |
| 英文摘要: |
| Abstract：Objective To investigate the inhibitory effect of recombinant human osteoprogerin-human serum album fusion protein(rhOPG-HAS)on the osteoclast precursor Raw264.7 cells under the simulated microgravity(SMG)．Methods Under the SMG condition，Raw264.7 cells and MC3T3-El cells were cultured. OPG activity in the MC3T3．El cell medium was detected with ELISA assay．Raw264.7 cells were induced to differentiate into osteoclasts by using macrophage colony stimulating factor(M-CSF)and soluble osteoclast differentiation factor(soluble receptor activator of nuclear factor-k13 ligand,sRANKL). The inhibitory effect of rhOPG-HSA on osteoclasts was identified using tartrate-resistant acid phosphatase (TRAP)staining．Semi-quantitative RT-PCR was used to detect the expression of TRAP in the osteoclasts．Results Under the SMG condition，OPG activity decreased in 72h(P<0.01)．TRAP positive cells reduced significantly(P<0.05)in rhOPG-HAS treated cultures，as compared with the negative control culture，in M-CSF and sRANKL induced Raw264.7 cells for 3d，4d，and 5d．RT-PCR results showed that the expression of TRAP decreased in the osteoclasts．Conclusion Under the SMG condition，rhOPG-HAS inhibited differentiation of the osteoclast precursor Raw264.7 cells． |
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