脂联素对人成骨细胞护骨素和核因子KB受体活化素配体表达的调节

Adiponectin regulates the expressions of osteopotegrin and receptor activator of NF-kappa B ligand in a time- and dose-dependent manners in human osteoblasts

目的探讨脂联素对人成骨细胞护骨素（OPG）和核因子xB受体活化素配体（RANKL）表达的作用。方法RT-PCR及Western印迹检测体外培养的人成骨细胞的脂联素受体（AdipoR）mRNA及AdipoR蛋白表达。分别用0、3、10μg／ml和30μg／ml脂联素干预人成骨细胞48h以观察剂量效应和分别用0、30μg／ml脂联素干预人成骨细胞0、12、24h和48h以观察时间效应，干预前后OPG、RANKL的mRNA及蛋白表达分别用实时聚合酶链反应（real—timePcR）和ELISA检测。结果（1）人成骨细胞可表达AdiopR1mRNA、AdiopR1蛋白和AdiopR2mRNA。（2）30μg／ml脂联素干预人成骨细胞12、24h或48h后，OPG的mRNA表达分别为0h的（69±6）％、（51士7）％和（26±8）％（Pd0．05），OPG蛋白表达分别为0h的（66±5）％、（32士8）％和（14士6）％（P％0．05）；RANKL的mRNA表达分别为0h的（180±12）％、（220士19）％和（316士14）％（P％0．05），可溶性RANKL（sRANKL）蛋白的表达分别为0h的（236±12）％、（272±9）％和（308±14）％（P〈O．05）。（3）3、10ug／ml或30ug／ml脂联素干预人成骨细胞48h时，OPG的tuRNA表达分别为对照组的（66±9）％、（44±10）％和（29±7）％（P〈O．05），OPG蛋白的表达分别为对照组的（52±9）％、（38±4）％和（17±4）％（P〈O．05）；RANKL的mRNA表达分另4为对照组的（168±6）％、（214±7）％和（314±11）％（P〈0．05），sRANKL蛋白的表达分别为对照组的（156±6）％、（196±8）％和（212±7）％（P〈0．05）。结论脂联素呈时间和剂量依赖性调节人成骨细胞OPG和RANKL的表达，可能是新的骨代谢调节因子。

Objective To investigate the action of adiponectin on the expressions of osteopotegrin（OPG） and receptor activator of NF-kappa B ligand（RANKL） in human osteoblasts. Methods Adiponectin receptor （Adipo R） in cultured osteoblasts was detected by retro-transcriptase polymerase chain reaction （RT-PCR） and Western blot. OPG and RANKL expressions were determined by using real-time PCR and enzyme linked immunosorbent assay （ELISA）. Human osteoblasts were incubated with 0 or 30 μg/ml of adiponectin for 0, 12, 24, 48 h to investigate the time-dependent effects, or with different doses of adiponectin （0, 3, 10, 30 μg/ml） for 48 h to investigate the dose-dependent effects. Results （1） Both AdipoR1 and AdipoR2 mRNA were expressed in normal human osteoblasts, but only AdipoR1 protein was detected in human osteoblasts. （2） After treating human osteoblasts with 30 μg/ml of adiponectin for 12, 24, 48 h, mRNA expressions of OPG were （69±6）%, （51±7）% or （26±8）% of the controls（0 h）（P〈0.05）,the levels of OPG were （66 ± 5 ） %, （ 32 ± 8） % or （ 14 ± 6 ） % of the controls, respectively （ P 〈 0.05 ） mRNA expressions of RANKL were （180± 12）%, （220 ± 19） % or （316 ± 14） % of the controls（ P〈 0.05）,the levels of sRANKL were （236±12）%,（272±9）% or （308±14）% of the controls,respectively（all P〈0. 05）. （3）After treating human osteoblasts with 3, 10, 30 μg/ml adiponectin for 48 h,mRNA expressions of OPG were （66±9）%, （44±10）%or （29±7）% of the controls（all P〈 0.05）, the levels of OPG were （ 52 ± 9） %, （38 ± 4） % or （ 17 ± 4） % of the controls, respectively （all P〈0.05）;mRNA expressions of RANKL were （168±6）%, （214±7）% or （314±11）% of the controls（P〈0.05）,the levels of sRANKL were （156±6）%, （196±8）%or （212±7）%of the controls, respectively （all P〈0. 05） . Conclusions Adiponectin could inhibit OPG expression in a time- and dose-dependent manners, and promote RANKL expression in the same way. These results may suggest a new physiological role of adiponectin on bone metabolism by regulating the expressions of OPG and RANKL in osteoblasts.