Tanshinone IIA promotes osteogenic differentiation potential and suppresses adipogenic differentiation potential of bone marrow mesenchymal stem cells

Wang,Wei; Wu,Hangqin; Feng,Shujing; Huang,Xingrui; Xu,Hao; Shen,Xinxin; Fu,Yajing; Fang,Shuchen

doi:10.3892/mmr.2024.13301

Tanshinone IIA promotes osteogenic differentiation potential and suppresses adipogenic differentiation potential of bone marrow mesenchymal stem cells

Authors:
- Wei Wang
- Hangqin Wu
- Shujing Feng
- Xingrui Huang
- Hao Xu
- Xinxin Shen
- Yajing Fu
- Shuchen Fang
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Affiliations: Department of Orthopedics, Hubei Provincial Hospital of Traditional Chinese Medicine, Wuhan, Hubei 430074, P.R. China, Department of Orthopedics, Wuyi County First People's Hospital, Wuyi, Zhejiang 321200, P.R. China, School of Exercise and Health, Shanghai University of Sport, Shanghai 200438, P.R. China, Department of Orthopedics, Suzhou Wujiang District Hospital of Traditional Chinese Medicine (Suzhou Wujiang District Second People's Hospital), Suzhou, Jiangsu 215003, P.R. China, Department of Anesthesiology, Renmin Hospital of Wuhan University, Wuhan, Hubei 430072, P.R. China
Published online on: August 7, 2024 https://doi.org/10.3892/mmr.2024.13301
Article Number: 177
Copyright: © Wang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Tanshinone IIA (Tan IIA) may have therapeutic effects on avascular necrosis of the femoral head (ANFH) by targeting bone marrow mesenchymal stem cells (BMSCs). The effect and underlying mechanism of Tan IIA on adipogenesis and osteogenesis ability of BMSCs remain to be elucidated. In the present study BMSCs were treated with osteogenic or adipogenic differentiation medium with or without Tan IIA under hypoxic environment. Osteogenic differentiation potential was evaluated by alkaline phosphatase (ALP) measurement, alizarin red staining and reverse transcription‑quantitative (RT‑q) PCR of osteogenic marker genes. Adipogenic differentiation potential was evaluated with oil red staining and RT‑qPCR of adipogenic marker genes. Detailed mechanism was explored by RNA‑seq and small molecular treatment during osteogenesis and adipogenesis of BMSCs. ALP level, mineralized nodules and expression level of osteogenic marker genes significantly increased following Tan IIA treatment during osteogenic differentiation of BMSCs. Lipid droplet and expression levels of adipogenic marker genes significantly decreased following Tan IIA treatment during adipogenic differentiation of BMSCs. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analyses of RNA‑seq data indicated increased Akt and TGFβ signaling following Tan IIA treatment. Further western blot assay confirmed that Tan IIA significantly activated Akt/cAMP response element‑binding protein signaling and TGFβ/Smad3 signaling. Application of Akti1/2 (an Akt inhibitor) significantly decreased the promotion effect of osteogenesis induced by Tan IIA, while the addition of SB431542 significantly reduced inhibition effect of adipogenesis caused by Tan IIA. Tan IIA could promote osteogenic differentiation potential of BMSCs by activating AKT signaling and suppress adipogenic differentiation potential of BMSCs by activating TGFβ signaling.

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1	Konarski W, Poboży T, Śliwczyński A, Kotela I, Krakowiak J, Hordowicz M and Kotela A: Avascular necrosis of femoral head-overview and current state of the art. Int J Environ Res Public Health. 19:73482022. View Article : Google Scholar : PubMed/NCBI
2	Rajpura A, Wright AC and Board TN: Medical management of osteonecrosis of the hip: A review. Hip Int. 21:385–392. 2011. View Article : Google Scholar : PubMed/NCBI
3	Sai Krishna MLV, Kar S, Kumar R, Singh H, Mittal R and Digge VK: The role of conservative management in the avascular necrosis of the femoral head: A review of systematic reviews. Indian J Orthop. 57:410–420. 2023.PubMed/NCBI
4	Li Z, Wang W, Xu H, Ning Y, Fang W, Liao W, Zou J, Yang Y and Shao N: Effects of altered CXCL12/CXCR4 axis on BMP2/Smad/Runx2/Osterix axis and osteogenic gene expressions during osteogenic differentiation of MSCs. Am J Transl Res. 9:1680–1693. 2017.PubMed/NCBI
5	Shapiro F, Connolly S, Zurakowski D, Menezes N, Olear E, Jimenez M, Flynn E and Jaramillo D: Femoral head deformation and repair following induction of ischemic necrosis: A histologic and magnetic resonance imaging study in the piglet. J Bone Joint Surg Am. 91:2903–2914. 2009. View Article : Google Scholar : PubMed/NCBI
6	Xu N, Liu H, Qu F, Fan J, Mao K, Yin Y, Liu J, Geng Z and Wang Y: Hypoxia inhibits the differentiation of mesenchymal stem cells into osteoblasts by activation of Notch signaling. Exp Mol Pathol. 94:33–39. 2013. View Article : Google Scholar : PubMed/NCBI
7	Benjamin S, Sheyn D, Ben-David S, Oh A, Kallai I, Li N, Gazit D and Gazit Z: Oxygenated environment enhances both stem cell survival and osteogenic differentiation. Tissue Eng Part A. 19:748–758. 2013. View Article : Google Scholar : PubMed/NCBI
8	Liu K, Ge H, Liu C, Jiang Y, Yu Y and Zhou Z: Notch-RBPJ pathway for the differentiation of bone marrow mesenchymal stem cells in femoral head necrosis. Int J Mol Sci. 24:62952023. View Article : Google Scholar : PubMed/NCBI
9	Wei B, Sun C, Wan H, Shou Q, Han B, Sheng M, Li L and Kai G: Bioactive components and molecular mechanisms of Salvia miltiorrhiza Bunge in promoting blood circulation to remove blood stasis. J Ethnopharmacol. 317:1166972023. View Article : Google Scholar : PubMed/NCBI
10	Sun K, Xue Y, Zhang X, Li X, Zhao J, Xu X, Zhang X and Yang F: Tanshinone I alleviates steroid-induced osteonecrosis of femoral heads and promotes angiogenesis: In vivo and in vitro studies. J Orthop Surg Res. 18:4742023. View Article : Google Scholar : PubMed/NCBI
11	Wu Y, Zhang C, Wu J, Han Y and Wu C: Angiogenesis and bone regeneration by mesenchymal stem cell transplantation with danshen in a rabbit model of avascular necrotic femoral head. Exp Ther Med. 18:163–171. 2019.PubMed/NCBI
12	Gao S, Liu Z, Li H, Little PJ, Liu P and Xu S: Cardiovascular actions and therapeutic potential of tanshinone IIA. Atherosclerosis. 220:3–10. 2012. View Article : Google Scholar : PubMed/NCBI
13	Xie J, Wang H, Song T, Wang Z, Li F, Ma J, Chen J, Nan Y, Yi H and Wang W: Tanshinone IIA and astragaloside IV promote the migration of mesenchymal stem cells by up-regulation of CXCR4. Protoplasma. 250:521–530. 2013. View Article : Google Scholar : PubMed/NCBI
14	Yuan P, Qin HY, Wei JY, Chen G and Li X: Proteomics reveals the potential mechanism of Tanshinone IIA in promoting the ex vivo expansion of human bone marrow mesenchymal stem cells. Regen Ther. 21:560–573. 2022. View Article : Google Scholar : PubMed/NCBI
15	Liu X, Niu Y, Xie W, Wei D and Du Q: Tanshinone IIA promotes osteogenic differentiation of human periodontal ligament stem cells via ERK1/2-dependent Runx2 induction. Am J Transl Res. 11:340–350. 2019.PubMed/NCBI
16	Kim HJ and Kim SH: Tanshinone IIA enhances BMP-2-stimulated commitment of C2C12 cells into osteoblasts via p38 activation. Amino Acids. 39:1217–1226. 2010. View Article : Google Scholar : PubMed/NCBI
17	Abbuehl JP, Tatarova Z, Held W and Huelsken J: Long-term engraftment of primary bone marrow stromal cells repairs niche damage and improves hematopoietic stem cell transplantation. Cell Stem Cell. 21:241–255.e6. 2017. View Article : Google Scholar : PubMed/NCBI
18	Davaapil H, McNamara M, Granata A, Macrae RGC, Hirano M, Fitzek M, Aragon-Martin JA, Child A, Smith DM and Sinha S: A phenotypic screen of Marfan syndrome iPSC-derived vascular smooth muscle cells uncovers GSK3β as a new target. Stem Cell Rep. 18:555–569. 2023. View Article : Google Scholar : PubMed/NCBI
19	Chiang J and Martinez-Agosto JA: Effects of mTOR inhibitors on components of the salvador-warts-hippo pathway. Cells. 1:886–904. 2012. View Article : Google Scholar : PubMed/NCBI
20	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(−Delta Delta C(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar : PubMed/NCBI
21	Reggio A, Rosina M, Palma A, Cerquone Perpetuini A, Petrilli LL, Gargioli C, Fuoco C, Micarelli E, Giuliani G, Cerretani M, et al: Adipogenesis of skeletal muscle fibro/adipogenic progenitors is affected by the WNT5a/GSK3/β-catenin axis. Cell Death Differ. 27:2921–2941. 2020. View Article : Google Scholar : PubMed/NCBI
22	Li B and Dewey CN: RSEM: Accurate transcript quantification from RNA-Seq data with or without a reference genome. BMC Bioinformatics. 12:3232011. View Article : Google Scholar : PubMed/NCBI
23	Wang L, Feng Z, Wang X, Wang X and Zhang X: DEGseq: An R package for identifying differentially expressed genes from RNA-seq data. Bioinformatics. 26:136–138. 2010. View Article : Google Scholar : PubMed/NCBI
24	Xie C, Mao X, Huang J, Ding Y, Wu J, Dong S, Kong L, Gao G, Li CY and Wei L: KOBAS 2.0: A web server for annotation and identification of enriched pathways and diseases. Nucleic Acids Res. 39((Web Server Issue)): W316–W322. 2011. View Article : Google Scholar : PubMed/NCBI
25	Guo W, Flanagan J, Jasuja R, Kirkland J, Jiang L and Bhasin S: The effects of myostatin on adipogenic differentiation of human bone marrow-derived mesenchymal stem cells are mediated through cross-communication between Smad3 and Wnt/beta-catenin signaling pathways. J Biol Chem. 283:9136–9145. 2008. View Article : Google Scholar : PubMed/NCBI
26	Tsurutani Y, Fujimoto M, Takemoto M, Irisuna H, Koshizaka M, Onishi S, Ishikawa T, Mezawa M, He P, Honjo S, et al: The roles of transforming growth factor-β and Smad3 signaling in adipocyte differentiation and obesity. Biochem Biophys Res Commun. 407:68–73. 2011. View Article : Google Scholar : PubMed/NCBI
27	Kim YJ, Hwang SJ, Bae YC and Jung JS: MiR-21 regulates adipogenic differentiation through the modulation of TGF-beta signaling in mesenchymal stem cells derived from human adipose tissue. Stem Cells. 27:3093–3102. 2009. View Article : Google Scholar : PubMed/NCBI
28	Cao H, Shi K, Long J, Liu Y, Li L, Ye T, Huang C, Lai Y, Bai X, Qin L and Wang X: PDGF-BB prevents destructive repair and promotes reparative osteogenesis of steroid-associated osteonecrosis of the femoral head in rabbits. Bone. 167:1166452023. View Article : Google Scholar : PubMed/NCBI
29	Kang H, Yang S and Lee J: Tauroursodeoxycholic acid enhances osteogenic differentiation through EGFR/p-Akt/CREB1 pathway in mesenchymal stem cells. Cells. 12:14632023. View Article : Google Scholar : PubMed/NCBI
30	Gao B, Deng R, Chai Y, Chen H, Hu B, Wang X, Zhu S, Cao Y, Ni S, Wan M, et al: Macrophage-lineage TRAP+ cells recruit periosteum-derived cells for periosteal osteogenesis and regeneration. J Clin Invest. 129:2578–2594. 2019. View Article : Google Scholar : PubMed/NCBI
31	Baker N, Sohn J and Tuan RS: Promotion of human mesenchymal stem cell osteogenesis by PI3-kinase/Akt signaling, and the influence of caveolin-1/cholesterol homeostasis. Stem Cell Res Ther. 6:2382015. View Article : Google Scholar : PubMed/NCBI
32	Han J, Chai Y, Zhang XY, Chen F, Xu ZW, Feng Z, Yan Q, Wen SB and Wu YK: Gujiansan ameliorates avascular necrosis of the femoral head by regulating autophagy via the HIF-1α/BNIP3 pathway. Evid Based Complement Alternat Med. 2021:66830072021. View Article : Google Scholar : PubMed/NCBI
33	Huang Z, Fu F, Ye H, Gao H, Tan B, Wang R, Lin N, Qin L and Chen W: Chinese herbal Huo-Gu formula for the treatment of steroid-associated osteonecrosis of femoral head: A 14-year follow-up of convalescent SARS patients. J Orthop Translat. 23:122–131. 2020. View Article : Google Scholar : PubMed/NCBI
34	Li ZR, Cheng LM, Wang KZ, Yang NP, Yang SH, He W, Wang YS, Wang ZM, Yang P, Liu XZ, et al: Herbal Fufang Xian Ling Gu Bao prevents corticosteroid-induced osteonecrosis of the femoral head-A first multicentre, randomised, double-blind, placebo-controlled clinical trial. J Orthop Translat. 12:36–44. 2017. View Article : Google Scholar : PubMed/NCBI
35	Li J, He C, Tong W, Zou Y, Li D, Zhang C and Xu W: Tanshinone IIA blocks dexamethasone-induced apoptosis in osteoblasts through inhibiting Nox4-derived ROS production. Int J Clin Exp Pathol. 8:13695–13706. 2015.PubMed/NCBI
36	Du K and Montminy M: CREB is a regulatory target for the protein kinase Akt/PKB. J Biol Chem. 273:32377–32379. 1998. View Article : Google Scholar : PubMed/NCBI
37	Tzavlaki K and Moustakas A: TGF-β signaling. Biomolecules. 10:4872020. View Article : Google Scholar : PubMed/NCBI