Analysis of the Potential Angiogenic Mechanisms of BuShenHuoXue Decoction against Osteonecrosis of the Femoral Head Based on Network Pharmacology and Experimental Validation

• Published in: Orthopaedic surgery Vol. 16; no. 3; pp. 700 - 717
• Main Authors: Luo, Di, Liu, Hao, Liang, Xue‐zhen, Yan, Wei, Ding, Chou, Hu, Cheng‐bo, Yan, De‐zhi, Li, Jin‐song, Wu, Ji‐biao
• Format: Journal Article
• Language: English
• Published: Melbourne John Wiley & Sons Australia, Ltd 01.03.2024; John Wiley & Sons, Inc; Wiley
• Subjects: Analysis; Angiogenesis; Bioinformatics; Bone marrow; Bones; Chinese medicine; Dexamethasone; Drug dosages; Experiments; Gene expression; Immunohistochemistry; Intervention; Laboratory animals; Molecular mechanisms of pharmacological action; Necrosis; Orthopedics; Osteonecrosis of the femoral head (ONFH); Pharmacology; Stem cells; Steroids; Traditional Chinese medicine (TCM); Von Willebrand factor; China; Osteonecrosis of the femoral head (ONFH); Molecular mechanisms of pharmacological action; Bioinformatics; Traditional Chinese medicine (TCM)
• ISSN: 1757-7853; 1757-7861
• DOI: 10.1111/os.13970

Objective Osteonecrosis of the femoral head (ONFH) is a common orthopedic disease with a high disability rate. The clinical effect of BuShenHuoXue decoction (BSHX) for ONFH is satisfactory. We aimed to elucidate the potential angiogenic mechanisms of BSHX in a rat femoral osteonecrosis model and bone marrow mesenchymal stem cells (BMSCs). Methods With in vivo experiments, we established the steroid‐induced osteonecrosis of the femoral head (SONFH) model using Sprague–Dawley (SD) rats (8‐week‐old). The rats were randomly divided into five group of 12 rats each and given the corresponding interventions: control, model (gavaged with 0.9% saline), BSHX low‐, medium‐ and high‐dose groups (0.132 3, 0.264 6, and 0.529 2 g/mL BSHX solution by gavage). After 12 weeks, haematoxylin and eosin (H&E) staining was preformed to evaluate rat osteonecrosis. the expression of angiogenic factors (CD31, VEGFA, KDR, VWF) in rat femoral head was detected by immunohistochemistry, qPCR and western blotting. In cell experiment, BMSCs were isolated and cultured in the femoral bone marrow cavity of 4‐week‐old SD rats. BMSCs were randomly divided into eight groups and intervened with different doses of BSHX‐containing serum and glucocorticoids: control group (CG); BSHX low‐, medium‐, and high‐dose groups (CG + 0.661 5, 1.323, and 2.646 g/kg BSHX gavage rat serum); dexamethasone (Dex) group; and Dex + BSHX low‐, medium‐, and high‐dose groups (Dex + 0.661 5, 1.323, and 2.646 g/kg BSHX gavaged rat serum), the effects of BSHX‐containing serum on the angiogenic capacity of BMSCs were examined by qPCR and Western blotting. A co‐culture system of rat aortic endothelial cells (RAOECs) and BMSCs was then established. Migration and angiogenesis of RAOECs were observed using angiogenesis and transwell assay. Identification of potential targets of BSHX against ONFH was obtained using network pharmacology. Results BSHX upregulated the expression of CD31, VEGFA, KDR, and VWF in rat femoral head samples and BMSCs (p < 0.05, vs. control group or model group). Different concentrations of BSHX‐containing serum significantly ameliorated the inhibition of CD31, VEGFA, KDR and VWF expression by high concentrations of Dex. BSHX‐containing serum‐induced BMSCs promoted the migration and angiogenesis of RAOECs, reversed to some extent the adverse effect of Dex on microangiogenesis in RAOECs, and increased the number of microangiogenic vessels. Furthermore, we identified VEGFA, COL1A1, COL3A1, and SPP1 as important targets of BSHX against ONFH. Conclusion BSHX upregulated the expression of angiogenic factors in the femoral head tissue of ONFH model rats and promoted the angiogenic capacity of rat RAOECs and BMSCs. This study provides an important basis for the use of BSHX for ONFH prevention and treatment. BSHX ameliorated glucocorticoid‐induced decrease in angiogenesis in the rat femoral head. BSHX‐containing serum promoted the migratory proliferative activity of rat aortic endothelial cells (RAOECs) and the angiogenic differentiation ability of bone marrow mesenchymal stem cells (BMSCs). VEGFA, COL1A1, COL3A1, and SPP1 were critical targets of BSHX in the prevention and treatment of osteonecrosis of the femoral head (ONFH).