Intervention with Compound Kidney-Invigorating Granules in a mouse model of osteoporosis: role of the TRIB3/beta-catenin axis

Abstract

BACKGROUND: Previous studies have shown that knockdown of β-catenin can inhibit the osteogenic differentiation of human bone marrow mesenchymal stem cells and reduce the expression of TRIB3. Serum containing Compound Kidney-Invigorating Granules can promote the expression of β-catenin and TRIB3 in human bone marrow mesenchymal stem cells, and induce human bone marrow mesenchymal stem cells to differentiate into osteogenic cells. OBJECTIVE: To further explore the mechanism of Compound Kidney-Invigorating Granules in a mouse model of osteoporosis based on the TRIB3/β-catenin axis. METHODS: 8-week-old female C57BL/6 mice were randomly divided into the following experimental groups: blank control, sham operation, model, and low-, medium-, and high-dose Compound Kidney-Invigorating Granules groups, and positive drug group. Except for the blank control and sham operation groups, bilateral ovariectomy was performed to establish an osteoporosis mouse model. One week after modeling, mice in the low-, medium-, and high-dose groups were intragastrically administered 7.05, 14.1, and 28.2 g/kg Compound Kidney-Invigorating Granules, respectively; the blank control, sham operation, and model groups received an equal volume of normal saline once daily; the positive control group received 1.53 mg/kg alendronate sodium once weekly. After 12 weeks of administration, Micro-CT was used to detect changes in femoral bone microarchitecture; hematoxylin-eosin staining and Masson staining were used to detect pathological changes in the femur; Western blot was used to detect the expression levels of TRIB3, β-catenin, alkaline phosphatase, and osteopontin in bone tissue. RESULTS AND CONCLUSION: Compared with the blank control and sham operation groups, the model group showed sparse bone trabeculae and significantly increased empty lacunae; the protein expression levels of TRIB3, β-catenin, alkaline phosphatase, and osteopontin were significantly decreased (P < 0.05). Compared with the model group, the medium- and high-dose Compound Kidney-Invigorating Granules groups showed more complete and regular bone trabeculae, and the protein expression levels of TRIB3, β-catenin, alkaline phosphatase, and osteopontin were significantly upregulated (P < 0.05). These results indicate that Compound Kidney-Invigorating Granules exert a therapeutic effect on osteoporosis model mice, suggesting that the formula may act through the TRIB3/β-catenin axis.

1. Introduction

Osteoporosis is a common skeletal disease characterized by reduced bone density and increased susceptibility to fractures, particularly in the aging population and postmenopausal women. The disease often progresses asymptomatically, leading to severe consequences such as fractures. The etiology of osteoporosis is multifactorial, involving complex interactions among genetic, hormonal, and environmental factors. When bone metabolic homeostasis is disrupted, excessive bone resorption or insufficient bone formation can trigger osteoporosis. In traditional Chinese medicine, osteoporosis is closely related to qi and blood deficiency and kidney deficiency, with kidney deficiency considered the main cause. The classic text Qianjin Yaofang states: 'Bone extreme is governed by the kidney; the kidney corresponds to bone, and bone and kidney are united.' The kidney is the foundation of innate constitution, and kidney deficiency with essence depletion is the core pathogenesis of osteoporosis.

Current commonly used drugs for osteoporosis, such as calcitonin and selective estrogen receptor modulators, can improve bone density and reduce fracture risk to some extent. However, numerous studies have shown that these drugs may have adverse effects during long-term use, such as hot flashes, nausea, and vomiting, making them unsuitable for prolonged application. Therefore, finding safe and sustainable drugs for osteoporosis is a major challenge. Compound Kidney-Invigorating Granules are a clinical empirical formula from the First Affiliated Hospital of Anhui University of Chinese Medicine. Previous basic research by our group found that serum containing Compound Kidney-Invigorating Granules can promote the proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells through the Wnt signaling pathway. Furthermore, Hu et al. found that knockdown of β-catenin inhibited the osteogenic differentiation of human bone marrow mesenchymal stem cells and reduced TRIB3 expression, while Compound Kidney-Invigorating Granules-containing serum could promote the expression of β-catenin and TRIB3, inducing osteogenic differentiation.