阿仑膦酸钠干预人骨髓基质细胞OPG/RANKL的体外实验研究  

Alendronate affects osteoprotegerin/receptor of nuclear factor κB-ligand activator expression in human bone marrow stroma cells in vitro

作　　者：王建忠[1] 王坤正[2] 周荣兴[1] 李晓东[1] 郭军[1] 杨铁翼[1] 吕慧成[1] 

机构地区：[1]内蒙古医学院第二附属医院创伤二科,呼和浩特010030 [2]西安交通大学医学院第二附属医院骨外科,710004

出　　处：《中华关节外科杂志（电子版）》2011年第5期56-60,共5页Chinese Journal of Joint Surgery(Electronic Edition)

基　　金：国家自然科学基金(81160228)

摘　　要：目的观察阿仑膦酸钠对人骨髓基质细胞中骨保护素（OPG）核/因子Kappa B受体活化因子配基（RANKL）mRNA表达的影响,探讨阿仑膦酸钠防治骨质疏松的相关机制。方法从24名20~30岁的健康男性志愿者髂前上棘处分别抽取10 m l骨髓,分离培养骨髓基质细胞,取50%融合P2代骨髓基质细胞,混匀后随机分为4组：高、中、低剂量阿仑膦酸钠组和对照组,高、中、低剂量组在细胞培养液中,分别加入1×10-7mol/L、1×10-8mol/L、1×10-9mol/L阿仑膦酸钠;对照组采用普通LG-DMEM培养,不进行特殊处理。采用半定量RT-PCR和W estern blot检测OPG、RANKL mRNA和蛋白表达并计算OPG/RANKL比率。结果在RT-PCR实验中,高、中、低阿仑膦酸钠组和对照组OPG mRNA/RANKL mRNA表达比分别为（8.77±1.16）、（6.68±1.25）、（4.86±0.79）和（0.58±0.13）;W estern blot蛋白印迹实验显示,高、中、低阿仑膦酸钠组和对照组OPG/RANKL蛋白表达之比分别为（1.18±0.47）、（1.09±0.56）、（0.82±0.32）和（0.25±0.12）。经统计学分析,在mRNA和蛋白水平,高、中、低阿仑膦酸钠组OPG/RANKL比值明显高于对照组,差异有统计学意义（P〈0.05）,三组阿仑膦酸钠组比较,差异无统计学意义（P〉0.05）。结论防治骨质疏松药阿仑膦酸钠可以增加骨髓基质细胞中骨保护素的表达,减少核因子Kappa B受体活化因子配基表达。Objective To evaluate the effects of alendronate on osteoprotegerin（OPG） and receptor of activator of nuclear factor κB-ligand（RANKL） expression in human bone marrow stroma cells（BMSCs） in vitro,and to investigate the related mechanisms of alendronate on prevention and treatment for osteoporosis.Methods 10ml bone marrow was taken at anterior superior iliac spine from 24 healthy male volunteers aged 20-30 years.BMSCs were isolated,cultured in vitro,and randomly divided into four groups： high,middle,low dose alendronate groups and control group.In high,middle and low dose groups,hMSCs culture fluid contained 1× 10-7mol/L,1× 10-8mol/L,1×10-9mol/L alendronate respectively.In control group,there was no special treatment but cultured hMSCs in LG-DMEM.Two weeks after treatment,the expressions of OPG and RANKL were evaluated by RT-PCR and Western blot.Results In RT-PCR,the ratios of OPG/RANKL in high,middle,low dose alendronate groups and control group were（8.77±1.16）,（6.68±1.25）,（4.86±0.79） and（0.58±0.13） respectively.In Western blot,the ratios of OPG/RANKL in high,middle,low dose alendronate groups and control group were（1.18±0.47）,（1.09±0.56）,（0.82±0.32） and（0.25±0.12） respectively.The ratio of OPG/RANKL was higher in alendronate group than it in control group（P0.01）,but there was no significant difference in three alendronate groups.Conclusion Alendronate may enhance OPG expression and inhibit RANKL expression of hMSCs in vitro.

关 键 词：阿仑膦酸盐 骨髓 骨保护素 核因子KappaB受体活化因子配基 

分 类 号：R580[医药卫生—内分泌]