Abstract
The apoptosis of mature osteocytes is the main factor causing damage to the microstructure of cortical bone in glucocorticoid-induced osteoporosis (GIOP). Our previous research found damaged areas and empty osteocytes lacunae in the tibial cortical bone of GIOP mice. However, the specific mechanism has not been clarified. Recently, a study showed that the quality of the cortical bone significantly increased by knocking out Notum, a gene encoding α/β hydrolase. However, it is not clear whether Notum affects cortical bone remodeling by participating in glucocorticoids (GCs)-induced apoptosis of osteocytes. The present study aimed to explore the correlation between Notum, osteocytes apoptosis, and cortical bone quality in GIOP. Prednisolone acetate was intragastrically administered to mice for two weeks. Histochemical staining was applied to evaluate changes in GIOP and Notum expression. Osteocytes were stimulated with prednisolone, and cell viability was assessed via CCK8. Hoechst 33342/PI staining, flow cytometry, RT-PCR, and western blot were used to detect osteocytes apoptosis, siRNA transfection efficiency, and expressions of pathway related factors. The results showed that the number of empty osteocytes lacunae increased in GIOP mice. TUNEL-stained apoptotic osteocytes and Notum immuno-positive osteocytes were also observed. Furthermore, prednisolone was found to promote Notum expression and osteocytes apoptosis in vitro. Knocking down Notum via siRNA partially restored osteocytes apoptosis and phosphoinositide 3-kinase (PI3K)/protein kinase B (AKT)/glycogen synthase kinase-3β (GSK3β)/β-catenin pathway. These findings showed GCs-induced osteocytes apoptosis by promoting Notum expression and inhibiting PI3K/AKT/GSK3β/β-catenin pathway. Thus, Notum might be a potential therapeutic target for the treatment of GIOP.
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Acknowledgements
This study was supported by the National Natural Science Foundation of China (No. 81972072) to Li M, the National Natural Science Foundation of China (Nos. 81970964, 81771108) to Guo J, the National Natural Science Foundation of China (No. 81800982) to Liu H, and the Construction Engineering Special Fund of “Taishan Young Scholars” of Shandong Province (No. tsqn 202103177) to Liu H.
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Conceptualization: CL, PY, BL; Methodology: CL, PY, HL, TH and JB; Formal analysis and investigation: CL, PY and ML; Writing—original draft preparation: CL; Writing—review and editing: CL; Data curation: CL and PY; Funding acquisition: ML, JG and HL; Resources: ML, JG and HS; Supervision: ML and JG; Project administration: ML; Software: CL; Validation: CL and ML; Visualization: CL. All authors made substantial contributions to conception and design, acquisition of data, or analysis and interpretation of data; took part in drafting the article or revising it critically for important intellectual content; agreed to submit to the current journal; gave final approval of the version to be published; and agree to be accountable for all aspects of the work.
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All experimental procedures complied with the ARRIVE guidelines and were conducted in accordance with the guidelines for the Care and Use of Laboratory Animals of the National Institutes of Health. And all animal experiments and the use of the cell line were approved by the ethics committee of School and Hospital of Stomatology, Shandong University (No. 20210116).
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Li, C., Yang, P., Liu, B. et al. Prednisolone induces osteocytes apoptosis by promoting Notum expression and inhibiting PI3K/AKT/GSK3β/β-catenin pathway. J Mol Histol 52, 1081–1095 (2021). https://doi.org/10.1007/s10735-021-10006-0
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DOI: https://doi.org/10.1007/s10735-021-10006-0