BushenHuoxue Formula Promotes Osteogenic Differentiation Via Affecting Hedgehog Signaling Pathway in Bone Marrow Stem Cells to Improve Osteoporosis Symptoms

Overview

Journal PLoS One

Specialties General Medicine
Science

Date 2023 Nov 29

PMID 38019761

Authors

Yuqi Chen

Zhiyong Wei

Hongxia Shi

Xin Wen

Yiran Wang

Rong Wei

Affiliations

Soon will be listed here.

Abstract

Background: The BushenHuoxue formula (BSHX) has been previously demonstrated to ameliorate osteoporosis, but the mechanisms underlying this phenomenon are currently unclear. The present study aims at investigating the mechanisms that BSHX induces osteogenesis.

Methods: We established an osteoporosis model in rats by bilateral ovariectomy and then treated the rats with an osteogenic inducer (dexamethasone, β-sodium glycerophosphate and Vitamin C) and BSHX. After that, bone marrow density and histopathological bone examination were evaluated by using HE staining and immunohistochemistry, respectively. We also assessed the differentiation of bone marrow mesenchymal stem cells (BMSCs) into osteoblasts by using immunofluorescence staining. ALP, BMP, and COL1A1 levels were determined by ELISA. We identified genes involved in pathogenesis of osteoporosis through Gene Expression Omnibus (GEO) database and subsequently selected Hedgehog signaling-related genes Shh, Ihh, Gli2, and Runx2 for assessment via qRT-PCR and ELISA, Western blotting. Network pharmacology analysis was performed to identify bioactive metabolites of BSHX.

Results: BSHX treatment in osteoporosis model rats promoted tightening of the morphological structure of the trabecular bone and increased the bone mineral density (BMD). BSHX also increased levels of osteoblast makers ALP, BMP, and COL1A1. Additionally, bioinformatics analysis of the GEO dataset showed that Hedgehog signaling pathway was involved in pathogenesis of osteoporosis, especially related genes Shh, Ihh, Gli2, and Runx2. Remarkably, BHSX upregulated these genes indispensably involved in the osteogenesis-related Hedgehog signaling pathway in both bone tissue and BMSCs. Importantly, we identified that quercetin was the active compounds that involved in the mechanism of BSHX-improved OP via affecting Hedgehog-related genes.

Conclusion: Our results indicate that BSHX promotes osteogenesis by improving BMSC differentiation into osteoblasts via increased expression of Hedgehog signaling-related genes Shh, Ihh, Gli2, and Runx2, and quercetin was the bioactive compound of BSHX.

Citing Articles

LINC01133 promotes the osteogenic differentiation of bone marrow mesenchymal stem cells by upregulating CTNNB1 by acting as a sponge for miR-214-3p.

Tang C, Huang L, Guo X, Wang G, Chen Z J Orthop Surg Res. 2024; 19(1):572.

PMID: 39285416 PMC: 11406849. DOI: 10.1186/s13018-024-05053-8.

CRYAB suppresses ferroptosis and promotes osteogenic differentiation of human bone marrow stem cells via binding and stabilizing FTH1.

Tian B, Li X, Li W, Shi Z, He X, Wang S Aging (Albany NY). 2024; 16(10):8965-8979.

PMID: 38787373 PMC: 11164484. DOI: 10.18632/aging.205851.

References

Jemtland R, Divieti P, Lee K, Segre G . Hedgehog promotes primary osteoblast differentiation and increases PTHrP mRNA expression and iPTHrP secretion. Bone. 2003; 32(6):611-20. DOI: 10.1016/s8756-3282(03)00092-9. View

Peng Z, Xu R, You Q . Role of Traditional Chinese Medicine in Bone Regeneration and Osteoporosis. Front Bioeng Biotechnol. 2022; 10:911326. PMC: 9194098. DOI: 10.3389/fbioe.2022.911326. View

Beloti M, Bellesini L, Rosa A . Purmorphamine enhances osteogenic activity of human osteoblasts derived from bone marrow mesenchymal cells. Cell Biol Int. 2005; 29(7):537-41. DOI: 10.1016/j.cellbi.2005.02.007. View

Gersbach C, Byers B, Pavlath G, Garcia A . Runx2/Cbfa1 stimulates transdifferentiation of primary skeletal myoblasts into a mineralizing osteoblastic phenotype. Exp Cell Res. 2004; 300(2):406-17. DOI: 10.1016/j.yexcr.2004.07.031. View

Aljohani S, Fliefel R, Ihbe J, Kuhnisch J, Ehrenfeld M, Otto S . What is the effect of anti-resorptive drugs (ARDs) on the development of medication-related osteonecrosis of the jaw (MRONJ) in osteoporosis patients: A systematic review. J Craniomaxillofac Surg. 2017; 45(9):1493-1502. DOI: 10.1016/j.jcms.2017.05.028. View

Dominici M, Le Blanc K, Mueller I, Slaper-Cortenbach I, Marini F, Krause D . Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement. Cytotherapy. 2006; 8(4):315-7. DOI: 10.1080/14653240600855905. View

Ozturk S, Cuneyit I, Altuntas F, Karagur E, Cort Donmez A, Ocak M . Resveratrol prevents ovariectomy-induced bone quality deterioration by improving the microarchitectural and biophysicochemical properties of bone. J Bone Miner Metab. 2023; 41(4):443-456. DOI: 10.1007/s00774-023-01416-z. View

Mo J, Yang R, Li F, He B, Zhang X, Zhao Y . Geraniin promotes osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) via activating β-catenin: a comparative study between BMSCs from normal and osteoporotic rats. J Nat Med. 2018; 73(1):262-272. DOI: 10.1007/s11418-018-1242-6. View

Yusuf A, Cummings S, Watts N, Feudjo M, Sprafka J, Zhou J . Real-world effectiveness of osteoporosis therapies for fracture reduction in post-menopausal women. Arch Osteoporos. 2018; 13(1):33. PMC: 5862911. DOI: 10.1007/s11657-018-0439-3. View

10.

Li L, Dong Q, Wang Y, Feng Q, Zhou P, Ou X . Hedgehog signaling is involved in the BMP9-induced osteogenic differentiation of mesenchymal stem cells. Int J Mol Med. 2015; 35(6):1641-50. DOI: 10.3892/ijmm.2015.2172. View

11.

Liu H, Xia X, Li B . Mesenchymal stem cell aging: Mechanisms and influences on skeletal and non-skeletal tissues. Exp Biol Med (Maywood). 2015; 240(8):1099-106. PMC: 4935286. DOI: 10.1177/1535370215591828. View

12.

Zhong L, Zhang Y, Li H, Fu H, Lv C, Jia X . Overexpressed miR-196a accelerates osteogenic differentiation in osteoporotic mice via GNAS-dependent Hedgehog signaling pathway. J Cell Biochem. 2019; 120(12):19422-19431. DOI: 10.1002/jcb.29166. View

13.

Kim K, Jung H, Kim C, Kim S, Cho H, Kim D . Evidence-based guidelines for fall prevention in Korea. Korean J Intern Med. 2017; 32(1):199-210. PMC: 5214733. DOI: 10.3904/kjim.2016.218. View

14.

Veronesi F, Torricelli P, Borsari V, Tschon M, Rimondini L, Fini M . Mesenchymal stem cells in the aging and osteoporotic population. Crit Rev Eukaryot Gene Expr. 2011; 21(4):363-77. DOI: 10.1615/critreveukargeneexpr.v21.i4.60. View

15.

Wu T, Hu E, Xu S, Chen M, Guo P, Dai Z . clusterProfiler 4.0: A universal enrichment tool for interpreting omics data. Innovation (Camb). 2021; 2(3):100141. PMC: 8454663. DOI: 10.1016/j.xinn.2021.100141. View

16.

Tian X, Zhu G, Wang J, Wang Q, Guan L, Tan Y . Study on the relation between tissues pathologies and traditional chinese medicine syndromes in knee osteoarthritis: Medical image diagnostics by preoperative X-ray and surgical arthroscopy. J Xray Sci Technol. 2016; 24(4):509-19. DOI: 10.3233/XST-160567. View

17.

Huycke T, Eames B, Kimmel C . Hedgehog-dependent proliferation drives modular growth during morphogenesis of a dermal bone. Development. 2012; 139(13):2371-80. PMC: 3367445. DOI: 10.1242/dev.079806. View

18.

Benisch P, Schilling T, Klein-Hitpass L, Frey S, Seefried L, Raaijmakers N . The transcriptional profile of mesenchymal stem cell populations in primary osteoporosis is distinct and shows overexpression of osteogenic inhibitors. PLoS One. 2012; 7(9):e45142. PMC: 3454401. DOI: 10.1371/journal.pone.0045142. View

19.

Li K, Zhang X, He B, Yang R, Zhang Y, Shen Z . Geraniin promotes osteoblast proliferation and differentiation via the activation of Wnt/β-catenin pathway. Biomed Pharmacother. 2018; 99:319-324. DOI: 10.1016/j.biopha.2018.01.040. View

20.

Levi B, James A, Nelson E, Brugmann S, Sorkin M, Manu A . Role of Indian hedgehog signaling in palatal osteogenesis. Plast Reconstr Surg. 2011; 127(3):1182-1190. PMC: 3078688. DOI: 10.1097/PRS.0b013e3182043a07. View