雷洛昔芬对小鼠成骨细胞中护骨素核因子-κB受体活化因子配体表达的影响

目的 雷洛昔芬对小鼠成骨细胞中护骨素(OPG)、核因子-κB(NF-κB)受体活化因子配体(RANKL)表达的影响. 方法 取出生24 h内的小鼠30只,无菌的条件下取出颅骨,应用酶消化法进行成骨细胞的培养,在培养成骨细胞的培养液中加入不同浓度的雷洛昔芬(0、1012、1010、109mol/L),应用反转录聚合酶链扩增(RT-PCR)法检测其对OPG/RANKL mRNA的表达及酶联免疫吸附法(ELISA)法检测OPG蛋白分泌的影响. 结果 OPG mRNA在实验组中表达较对照组强,并且不同浓度组间的比较,差异有统计学意义(均为P<0.05),1010.mol/L组较10～mol/L、1012mol/L组的表达明显增强;RANKL mRNA在实验组的表达均较对照组弱,其组间的差异亦有统计学意义(P<0.01).并且随着雷洛营芬浓度的增加,RANKL mRNA的表达明显减弱,呈现明显的浓度依赖性.在试验组小鼠成骨细胞培养液中OPG浓度(10-9mol/L组为3.017±0.459,1010mol/L组为3.981±0.762,1012mol/L组为2.864±0.416)较对照组(2.106±0.316)增高,差异有统计学意义(P<0.05). 结论 雷洛昔芬能促进成骨细胞的中OPG的mRNA的表达及其蛋白的分泌,同时抑制RANKL的mRNA的表达.

The effects of raloxifene on OPG/ RANKL expression in mouse osteoblasts

Objective To investigate the effects of raloxifene on osteoprotegerin (OPG)/receptor activator nuclear factor kappa B ligand (RANKL) expression in mouse osteoblasts.Methods Sterile calvaria of mouse was taken from 30 newborn mice, and the osteoblasts were separated by enzyme digestion methods. Raloxifene in different concentrations (0,10<'-12>, 10<'-10>, 10<'-9>mol/L) were administrated into culture medium. The OPG/RANKL mRNA expression and OPG protein secretion were examined by RT-PCR and ELISA methods respectively. Results OPG mRNA expression in osteoblasts after raloxifene treatment was significantly higher than that in control group(P<0.05), and compared to 10<'-9> mol/L and 10<'-12> mol/L groups, it was significantly increased in 10<'-10> mol/L group. RANKL mRNA expression in osteoblasts after raloxifene treatment was significantly lower than that in control group(P<0.01), and the effect showed a dose- dependent manner. Compared to the control group, OPG protein secretion of osteoblasts was promoted by raloxifene treatment (10<'-9> mol/L:3.017±0.459;10<'-10> mol/L: 3. 981±0.762;10<'-12> mol/L : 2.864±0.416; control: 2.106±0.316, P<0.05). Conclusions Raloxifene can increase OPG mRNA expression, promote OPG protein secretion and inhibit RANKL mRNA expression in osteoblasts.