Effects of Loganin on Osteoblasts and Coupling Activities between Osteoblasts and Osteoclasts
WANG Yu, YANG Lintao, SUN Boyu, LIU Yu, LAN Tian, TSENG Yiider*
Building on its effect in promoting osteoblast differentiation, this study investigated how Loganin influences the coupling between osteoblasts and osteoclasts. To investigate the effects of Loganin on the coupling activities between osteoblasts and osteoclasts based on the promotion of osteoblast differentiation by Loganin. Using mouse embryonic osteoblast precursor cells (MC3T3-E1) and mouse mononuclear macrophage leukemia cells (RAW264.7) as experimental models, the effects of Loganin on MC3T3-E1 cell viability were assessed using the CCK-8 assay. The ALP (alkaline phosphatase) staining, ALP activity assay, and alizarin red staining were applied to evaluate the Loganin’s impact on osteoblast differentiation. The qRT-PCR was utilized to detect the transcriptions of osteoblast differentiation-related genes, and Western blot were applied to analyze the expressions of related proteins. The effects of Loganin on the coupling activities between osteoblasts and osteoclasts were examined using a co-culture system. The formation of osteoclasts and their resorption function were assessed by TRAP staining and F-actin staining; and the expression of osteoclast differentiation-related genes was detected by qRTPCR. The results showed that Loganin significantly enhanced the viability, ALP activity, and mineralized nodule formation of MC3T3-E1 cells within a certain concentration range (0.1 μmol/L and 1 μmol/L) in a concentrationdependent manner. Moreover, Loganin (1 μmol/L) significantly upregulated the gene expression levels of osteoblast differentiation markers Runx2, OSX, and OPG, as well as the protein expression levels of Runx2, OPG, and ALP, while downregulating the gene and protein expression levels of RANKL. In co-culture experiments, the conditioned medium from the Loganin group significantly inhibited the formation of osteoclasts and F-actin rings and downregulated the expression of CTSK and TRAP genes. Therefore, Loganin not only promotes osteoblast differentiation but also affects the coupling function between osteoblasts and osteoclasts by upregulating OPG and downregulating RANKL, thereby inhibiting the generation and resorption function of osteoclasts and regulating bone metabolism balance.
CN
EN