| Objective To study the mechanism of Gubenzenggu granules in preventing and treating osteoporosis, and to reveal the molecular mechanism of Gubenzenggu granules inhibiting osteoporosis through Wnt/β-catenin signaling pathway. Methods A total of 66 rats were randomly divided into sham operation group (13 rats) and modeling group (53 rats). The modeling group underwent bilateral back ovariectomy to prepare the model, while the sham operation group did not remove the ovaries. Each group was weighed 8 weeks after membrane construction, and 3 rats were randomly selected for bone mineral density detection. The modeling group was randomly divided into model group, Gubenzenggu granule high, medium and low dose groups and positive drug group, with 10 in each group. Gubenzenggu granule was given gavage at different concentrations of 24.4 g/kg, 12.2 g/kg and 6.1 g/kg at high, medium and low doses, respectively. The positive drug group was given estradiol at 0.09 mg/kg concentration by gavage, and the sham operation group and model group were given normal saline gavage. The daily gavage volume of rats was 2ml. The behavior of rats in each group was recorded every week. After 12 weeks of intervention, the femurs were sacrificed, and bone morphology was stained with HE, and the protein contents of Wnt7b, TCF3 and GSK3β in the femur tissues were detected by IHC. The mRNA and protein levels of Wnt7b, TCF3 and GSK3β in femur tissues were determined by RT-PCR and WB. Results HE staining showed that the bone structure of the sham operation group was normal, the trabecular bone thickness was uniform, the structure was complete, the trabecular connection was good, and the morphology of fat cells and bone cells was clear. In the model group, bone trabeculae of femur became thiner and irregular with a disorganized mesh structure, and some bone trabeculae were obviously elongated or fractured, with incomplete connection between the two. After the intervention of Gubenzenggu Granule, the bone trabecular structure of rats in the Gubenzenggu granule treatment group gradually tended to mature, the thickness increased, and the bone cells increased. The bone trabeculae were closely connected to the surrounding tissues, and the continuity gradually improved. The overall structure gradually improved. Compared with sham operation group, the expressions of Wnt7b and TCF3 in femur tissue of model group were significantly decreased (P<0.01), while the expression of GSK3β was significantly increased (P<0.01). Compared with model group, the expression levels of Wnt7b and TCF3 in positive drug group, Gubenzenggu granule high dose and medium dose groups were increased (P<0.05), and the expression levels of GSK3β were decreased (P<0.05). Conclusion Gubenzenggu granules can regulate the Wnt/β-catenin signaling pathway, up-regulate the expression of Wnt7b and TCF3 to promote bone formation, down-regulate the expression of GSK3β to prevent bone over-absorption. |