Objective To explore the effect of serum containing Zuogui Pill on the mRNA expression of key factors in the Hedgehog-GLi pathway and bone resorption function in an osteoblast-osteoclast-bone slice co-culture system. Methods An osteoblast-osteoclast-bone slice co-culture system was established. The experiment included blank control group, sham operation group, OVX group, positive control group, Zuogui Pill group, agonist group, and agonist + Zuogui Pill group. Serum containing drugs was prepared for the blank control group, sham operation group, OVX group, positive control group, and Zuogui Pill group using 50 female SD rats. Real-time PCR was used to measure the mRNA levels of Ihh, Ptch, Smo, GLi1, OPG, and RANKL in osteoblasts Tartrate-resistant acid phosphatase staining was employed to examine osteoc bone resorption function. Results Compared with the sham operation group, the OVX group exhibited a significant upregulation in mRNA expression levels of Ihh, Ptch, Smo, GLi1, while exhibiting significant decreases in OPG mRNA expression. In contrast, RANKL mRNA expression was significantly increased, resulting in a increase in the RANKL/OPG ratio. Compared with the OVX group, the mRNA expression levels of Ihh, Ptch, Smo, GLi1 in the positive control group and Zuogui Pill group were significantly downregulated, Meanwhile, there was a significant increase in OPG mRNA expression and a significant decrease in RANKL expression, which led to a significant decrease in the RANKL/OPG ratio. After adding Shh recombinant protein agonist to the OVX group, the mRNA expression of key HG pathway factors was significantly increased compared with the OVX group. In the agonist + Zuogui Pill group, the mRNA expression levels of Ihh, Ptch, Smo, GLi1 and RANKL/OPG ratio were significantly lowered compared to the agonist group. Moreover, the bone resorption pit area was significantly higher in the OVX group than in the sham group, compared with the OVX group, the positive control group and Zuogui Pill group showed a significant reduction. Conclusion Serum containing Zuogui Pill can inhibit the activity of key factors in the Hedgehog-GLi pathway, thereby suppressing the increase in bone resorption caused by OVX. |