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 rhOPG-HSA on osteoclast precursor Raw264.7 cells under simulated microgravity

作　　者：刘俊丽[1] 宋淑军[1] 司少艳[1] 冯凯[1] 刘军连[1] 赵刚[1] 张建中[1] 

机构地区：[1]解放军第306医院,北京100101

出　　处：《中国骨质疏松杂志》2011年第6期467-470,共4页Chinese Journal of Osteoporosis

基　　金：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

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RT-PCR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。Objective To investigate the inhibitory effect of recombinant human osteoprogerin-human serum album fusion protein （rhOPG-HAS） on the osteoclast precursor Raw264. 7 ceils under the simulated microgravity （SMG）. Methods Under the SMG condition, Raw264.7 cells and MC3T3-E1 cells were cultured. OPG activity in the MC3T3-E1 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-kβ 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-HSA inhibited differentiation of the osteoclast precursor Raw264.7 cells.

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 

分 类 号：R336[医药卫生—人体生理学]