| Objective To investigate the effect and mechanism of Wen Shen Yi Sui Decoction(WSYSD)-containing serum on RANKL-induced osteoclast (OC) differentiation. Methods Bone marrow mononuclear cells(BMMs) were intervened with different mass fractions of WSYSD-containing serum, and the drug-containing serum without cytotoxicity was screened by CCK-8 method for subsequent experiments. The control group, RANKL-induced group, and RANKL+WSYSD-containing serum of different mass fractions were set up, and after the corresponding interventions in each group, TRAP staining and toluidine blue staining were used to observe the OC and the formation of bone trap fossa, RT-qPCR and Western blot were used to detect the expression of mRNA and protein of TRAP, c-FOS, and CTR, and Western blot was used to detect the phosphorylation levels of ERK, JNK and p38 proteins in MAPK pathway. Results WSYSD-containing serum with mass fractions of 5%, 10%, and 15% did not reduce cell survival compared with the control group (P>0.05). Compared with the RANKL-induced group, the WSYSD-containing serum group (5%, 10%, and 15%) inhibited OC differentiation and bone trap formation, and decreased TRAP, c-FOS, and CTR mRNA expression, with mass fraction-dependent effects, with 15% being the best (P<0.01, P<0.01, and P<0.01). Compared with the RANKL-induced group, the WSYSD-containing serum group (15%) inhibited TRAP, c-FOS, and CTR protein expression, with a time-dependent effect, which was most pronounced on the 5th d (P<0.01). In addition, compared with the RANKL-induced group, the WSYSD-containing serum group (15%) inhibited the phosphorylation of ERK, JNK, and p38 proteins in the MAPK pathway, and the effect was also time-dependent, with the most obvious effect at the 60th min (P<0.01). Conclusion WSYSD-containing serum inhibited RANKL-induced OC differentiation, and the mechanism of action may be related to the inhibition of MAPK pathway activation. |