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Glucocorticoids Enhanced Osteoclast Autophagy Through the PI3K/Akt/mTOR Signaling Pathway

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Abstract

Autophagy is an evolutionarily conserved dynamic process and present in variety of cells at basal levels to maintain homeostasis and to promote cell survival in response to stresses. The early bone loss with excessive glucocorticoids (GCs) was reported to be related with the extension of the life span of osteoclasts. However, the connection between GCs induced bone loss and osteoclast autophagy remains to be elucidated. Autophagy was detected in a Dexamethasone (Dex) induced osteoporotic mice model and primary osteoclast cultures by autophagosome detection kit, and autophagy-related proteins were assayed by Western blotting and Immunostaining. The bone morphology was examined by micro-CT and TRAP staining. The trabecular bone micro-architecture was deteriorated, and the osteoclast number and spread area were increased in the Dex-treated mice compared with the control group (P < 0.01). Meanwhile, autophagy in pre-osteoclasts was increased in mice under Dex administration evidenced by the increased number of autophagosome and up-regulation of autophagy-related protein levels. Further, the enhanced autophagy under Dex treatment was verified in primary cultured osteoclasts, as shown by the increased levels of Beclin 1 and LC3-II/LC3-I and the autophagy complex formation members including Atg1, Atg13, and Atg7. However, the expressions of PI3K, p-Akt and p-mTOR in primary cultured osteoclasts were inhibited under Dex induced autophagy. Using the selective PTEN inhibitor SF1670 to activate the PI3K/Akt/mTOR pathway reversed this osteoclast autophagy under Dex treatment. Our study suggests that osteoclast autophagy was enhanced in glucocorticoids induced bone loss, and the PI3K/Akt/mTOR signaling pathway mediated the increased autophagy in primary cultured osteoclasts under glucocorticoids treatment.

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Acknowledgements

This work was supported by the National Nature Science Foundation of China (Grant No. 81201424), the Projects of Shanghai Municipal Health Bureau (Grant No. 20124332), Shanghai Clinical Medical Center (Grant No. 2017ZZ01023) and Shanghai Municipal Key Clinical Specialty.

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  1. Shanghai Key Laboratory of Orthopaedic Implants, Department of Orthopaedic Surgery, Shanghai Ninth People’s Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, 200011, People’s Republic of China

    Lingjie Fu, Wen Wu, Xiaojiang Sun & Pu Zhang

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All authors contributed to the study conception and design. Material preparation, data collection and analysis were performed by LF, WW and XS. The first draft of the manuscript was written by LF and all authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.

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Correspondence to Pu Zhang.

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All procedures performed in studies involving animals were in accordance with the ethical standards of the institution at which the studies were conducted and ethical approval was obtained from Ethics Committee and the Institutional Animal Care and Use Committee of Shanghai Ninth People’s Hospital.

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The Animal Care and Use Committee of Shanghai Ninth People’s Hospital approved the use of animals in this study. All applicable institutional and/or national guidelines for the care and use of animals were followed.

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Fu, L., Wu, W., Sun, X. et al. Glucocorticoids Enhanced Osteoclast Autophagy Through the PI3K/Akt/mTOR Signaling Pathway. Calcif Tissue Int 107, 60–71 (2020). https://doi.org/10.1007/s00223-020-00687-2

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