独活寄生汤含药血清对成骨细胞OPG/RANKL mRNA表达的影响

Effect of Serum Containing Duhuo Jisheng Decoction Substances on OPG/RANKLmRNA Expression of Osteoblasts

目的:检测独活寄生汤含药血清对成骨细胞OPG/RANKL mRNA表达的影响,从分子生物学层面探究独活寄生汤治疗骨质疏松症的机理。方法:原代体外培养SD乳大鼠成骨细胞,分别用生理盐水血清对照组、独活寄生汤含药血清低浓度,中浓度,高浓度作用于成骨细胞,采用碱性磷酸酶(ALP)检测成骨细胞活性,RT-PCR法检测独活寄生汤含药血清对成骨细胞OPG、RANKL mRNA表达的影响。结果:独活寄生汤能增加成骨细胞内碱性磷酸酶的活性,RT-PCR结果显示不同浓度的独活寄生汤含药血清能促进OPGmRNA的表达,并且抑制成骨细胞RANKL mRNA的表达。结论:独活寄生汤含药血清一则可以通过抑制RANKL分泌从而降低破骨细胞活性;二则促进成骨细胞分泌OPG,使之与RANKL结合增多,进而使破骨细胞活性降低,从而达到治疗骨质疏松的目的。

Objective：To investigate the effects of serum containing substances of Duhuo Jisheng Decoction on mRNA expression of osteoprotegerin （OPG） and receptor activator of NF - KB ligand （RANKL） in osteoblasts ,and to study the mechanism of action of Duhuo Jisheng Decoction for osteoporosis. Methods：Osteoblasts were obtained from new born rats＇calvaria by eollagenase -trypsin digestion. Four groups were included in the experiment, in situ hybridization method was used to detect the mRNA expression of OPG and RANKL. Results ： Compared with the normal serum group, the mRNA expression of OPG in osteoblasts in the serum of Duhuo Jisheng Decoction was increased significantly, while the mRNA expression of RANKL was decreased remarkably. Conclusion ： The serum containing Duhuo Jisheng Decoction could not only inhibit the secretion of RANKL to repress the activity of osteoclast but also stimulate osteoblasts to secrete OPG, then the combination of OPG and RANKL was increased,which could reduce the activity of RANKL and reduce the activity of the bone resorbing activity of obsteocIasts consequently. It may be one of the mechanism of Duhuo Jisheng Decoction for treating osteoporosis.