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Activating Wnt/β-catenin signaling by autophagic degradation of APC contributes to the osteoblast differentiation effect of soy isoflavone on osteoporotic mesenchymal stem cells

Acta Pharmacologica Sinica volume 44, pages 1841–1855 (2023)Cite this article

Abstract

The functional role of autophagy in regulating differentiation of bone marrow mesenchymal stem cells (MSCs) has been studied extensively, but the underlying mechanism remains largely unknown. The Wnt/β-catenin signaling pathway plays a pivotal role in the initiation of osteoblast differentiation of mesenchymal progenitor cells, and the stability of core protein β-catenin is tightly controlled by the APC/Axin/GSK-3β/Ck1α complex. Here we showed that genistein, a predominant soy isoflavone, stimulated osteoblast differentiation of MSCs in vivo and in vitro. Female rats were subjected to bilateral ovariectomy (OVX); four weeks after surgery the rats were orally administered genistein (50 mg·kg−1·d−1) for 8 weeks. The results showed that genistein administration significantly suppressed the bone loss and bone-fat imbalance, and stimulated bone formation in OVX rats. In vitro, genistein (10 nM) markedly activated autophagy and Wnt/β-catenin signaling pathway, and stimulated osteoblast differentiation in OVX-MSCs. Furthermore, we found that genistein promoted autophagic degradation of adenomatous polyposis coli (APC), thus initiated β-catenin-driven osteoblast differentiation. Notably, genistein activated autophagy through transcription factor EB (TFEB) rather than mammalian target of rapamycin (mTOR). These findings unveil the mechanism of how autophagy regulates osteogenesis in OVX-MSCs, which expands our understanding that such interplay could be employed as a useful therapeutic strategy for treating postmenopausal osteoporosis.

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Fig. 1: Genistein treatment prevented trabecular bone loss and promoted cancellous bone formation in ovariectomized rats.
Fig. 2: Genistein treatment reversed trabecular bone loss and abnormal autophagy induced by ovariectomy.
Fig. 3: Effects of genistein on proliferation, lineage commitment and autophagy of OVX-MSCs.
Fig. 4: Autophagy induction is required for genistein-mediated osteoblastic differentiation and Wnt/β-catenin signaling activation.
Fig. 5: Genistein-induced autophagy activates canonical Wnt/β-catenin signaling pathway.
Fig. 6: Genistein upregulates active β-catenin protein level through autophagic degradation of APC.
Fig. 7: TFEB upregulation is involved in genistein-activated autophagy.
Fig. 8: Effects of genistein on proliferation, osteoblast differentiation and autophagic activity of healthy-MSCs.

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Acknowledgements

This study was supported by the National Natural Science Foundation of China (grant number, 32000976 and 82071134).

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Authors and Affiliations

  1. Department of Oral Surgery, Shanghai Ninth People’s Hospital, Shanghai Jiao Tong University School of Medicine; College of Stomatology, Shanghai Jiao Tong University; National Center for Stomatology; National Clinical Research Center for Oral Diseases; Shanghai Key Laboratory of Stomatology; Shanghai Research Institute of Stomatology, Shanghai, 200001, China

    Jing Ge, Ye-jia Yu, Jia-yi Li, Meng-yu Li, Si-mo Xia, Shao-yi Wang & Chi Yang

  2. Department of Pastic and Reconstructive Surgery, Shanghai Ninth People’s Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, 200001, China

    Ke Xue

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Contributions

JG designed the experiments and wrote the manuscript. JG, YJY, JYL, and MYL performed most of the experiments. KX prepared the bone specimens and performed the histology. SMX generated the OVX rat model. SYW contributed to analysis and interpretation of data. CY supervised the study, and revised manuscript critically for important intellectual content. All authors have read and approved the manuscript.

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Correspondence to Shao-yi Wang or Chi Yang.

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Ge, J., Yu, Yj., Li, Jy. et al. Activating Wnt/β-catenin signaling by autophagic degradation of APC contributes to the osteoblast differentiation effect of soy isoflavone on osteoporotic mesenchymal stem cells. Acta Pharmacol Sin 44, 1841–1855 (2023). https://doi.org/10.1038/s41401-023-01066-x

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