蛇床子素对大鼠成骨细胞中OPG和RANKL基因mRNA表达的影响

Effects of osthole on expressions of OPG and RANKL in rat osteoblasts

目的 观察蛇床子素(OST)对体外培养的大鼠成骨细胞中OPG中RANKL基因表达的影响。方法 新生SD大鼠颅骨成骨细胞培养,设空白对照组和用不同浓度蛇床子素培养液(10-7,10-6,10-5mol／L)培养处理的大鼠成骨细胞为实验组,未经处理的细胞为空白对照组,分别提取48 h和7 d的细胞总mRNA,用RT-PCR技术分析OPG／RANKL的mRNA表达情况进行半定量比较。结果48h两基因都已有表达,相对空白对照组,OST上调OPG mRNA的表达(P<0.05),但对RANKL的表达无显著影响,OPG／RANKL比值变大;7 d时OPG和RANKL表达都有增加,OST上调OPG的表达(P<0.01),OST在10-5mol／L下调RANKL的表达(P<0.05),OPG／RANKL比值增大。结论OST可以增加大鼠成骨细胞OPG的表达,同时轻微抑制RANKL的表达,早期对RANKL的表达影响不大。

Objective To study the effects of osthole(OST) on expression of OPG and RANKL mRNA in rat osteoblasts. Methods Calvariae were obtained from Sprague-Dawley rats(24h after birth), bones samples were processed by collagenase/trypsin digestion, and the osteoblasts were cultured in vitro. 48h and 7 days after addition of osthole (0, 10-7,10-6,10-5 mol·L-1) to the culture medium of osteoblasts, total mRNA was prepared from osteoblasts, and mRNA expression of OPG and RANKL were deleted by RT-PCR. Results 48h after treatment with OST, the genes of OPG and RANKL were both expressed. Compared with the controls, OST increased the expression of OPG mRNA but had no effect on the expression of RANKL mRNA, the OPG/RANKL ratio being increased markedly in the treatment group. The mRNA expressions of OPG and RANKL were both increased 7 days after treatment with OST. OST at 10-5 mol·L-1 increased the expression of OPG mRNA, but deceased the expression of RANKL, the OPG/RANKL ratio in the treatment group was also increased, compared with the control. Conclusions Osthol stimulates the OPG mRNA expression of osteoblasts and decreases RANKL mRNA level. And osthol has no effect on the expresion of RANKL mRNA at the early stage of the experiment.