Effect of serum containing Yishen Juanbi method on OPG/RANKL/RANK signaling pathway in osteoblasts and osteoclasts of rats

Objective To explore the effect of serum containing Yishen Juanbi method on the signal pathway of bone metabolism. Methods SD rats were randomly divided into control groups and Yishen Juanbi Decoction groups. The two group rats were given saline solution and Yishen Juanbi Decoction by gavage, respectively, and the serum containing Yishen Juanbi decoction was collected. The osteoblasts were isolated and verified by Van kossa staining and ALP staining, and the osteoclasts were isolated and verified by TRAP and toluidine blue staining. Both cell types were separated into four groups: control group, low, medium and high doses of serum containing Yishen Juanbi decoction groups. The immunoblotting assay was utilized to measure the expression of osteoprotegerin(OPG), receptor activator of nuclear factor-κB ligand(RANKL) and osteoclast receptor activator of nuclear factor-κB(RANK). Results Compared to control group, the expression of OPG and RANKL in osteoblasts induced by low, medium and high concentrations of serum containing Yishen Juanbi decoction were 97%(P=0.710), 122%(P=0.005) and 155%(P<0.001) respectively, and the expression of RANKL was 81%(P<0.001), 63%(P<0.001) and 53%(P<0.001) respectively. The results indicated that the expression of OPG was positively correlated with the concentration of serum and the expression of RANKL was negatively correlated with the concentration of serum. Meanwhile, the expression of RANK in osteoclasts was regulated to 75%(P<0.001), 50%(P<0.001) and 46%(P<0.001) compared to the control group, suggesting that the expression of RANK was negatively correlated with the concentration of serum. Conclusion The serum containing Yishen Juanbi decoction may play a prevention role in the process of bone loss by regulating OPG/RANKL/rank signal axis.