摘要
目的 :研究护骨素配基 (OPGL)反义RNA对成骨细胞OPGL和护骨素 (OPG)mRNA表达及破骨细胞样细胞 (OLC)生成的影响。方法:构建小鼠OPGLcDNA反义表达载体 ,转染小鼠成骨细胞 ,筛选稳定表达细胞 ,通过RT -PCR研究其OPGL和OPGmRNA表达的变化。将转染有OPGL反义基因的成骨细胞与小鼠骨髓细胞共培养 ,观察OPGL反义RNA对OLC生成的影响。结果:转染有反义OPGLcDNA的成骨细胞OPGLmRNA表达水平显著降低 ,OPGmRNA表达不受影响。其诱导的OLC数目显著减少。结论:OPGL反义RNA抑制了OPGL基因的表达 ,使得成骨细胞促进破骨细胞形成、分化的能力减弱。
Objective:To investigate the effect of antisense RNA of osteoprotegerin ligand(OPGL)on osˉteoclastogenesis.Methods:The cDNA fragment of murine OPGL,obtained from RT-PCR,was cloned into a eukaryotic expression vector pcDNA3and then the vector was transfected into murine osteoblasts with lipofecˉtamine.In the presence of G418,cells were screened out that express the antigene permenantly.Then RT-PCR was carried out to study the effect of the antigene on the mRNA expression of OPGL and osteoprotegerin(OPG)in osteoblasts.Thereafter,the transfected cells and primary murine bone marrow cells were cocultured and the effect of the antigene was studied on osteoclastogenesis.Results:The mRNA levels of OPGL in osˉteoblasts were elevated after the transfection with the antisense expression vector,although no significant change was found for OPG.The potential of the osteoblasts transfected with the antigene to induce osteoclasˉtogenesis was significantly reduced.Conclusion:The antisense gene targeted to OPGL might confer a possible method in treating certain metabolic disorders of bone characterized by accelerated bone resorption.
出处
《天津医药》
CAS
北大核心
2004年第1期31-34,共4页
Tianjin Medical Journal
基金
国家自然科学基金资助项目 (项目编号 :39770669)
