MiR-27a-3p Targets GLP1R to Regulate Differentiation, Autophagy, and Release of Inflammatory Factors in Pre-Osteoblasts via the AMPK Signaling Pathway

Zhi Zeng; Liangyu Fei; Juntao Yang; Jun Zuo; Zelin Huang; Hao Li

doi:10.3389/fgene.2021.783352

MiR-27a-3p Targets GLP1R to Regulate Differentiation, Autophagy, and Release of Inflammatory Factors in Pre-Osteoblasts via the AMPK Signaling Pathway

Front Genet. 2022 Jan 5:12:783352. doi: 10.3389/fgene.2021.783352. eCollection 2021.

Authors

Zhi Zeng¹, Liangyu Fei², Juntao Yang³, Jun Zuo³, Zelin Huang³, Hao Li³

Affiliations

¹ Department of Joint Surgery, The Second Affiliated Hospital, Hengyang Medical School, University of South China, Hengyang, China.
² Department of Nephrology and Rheumatology, The Affiliated Nanhua Hospital, Hengyang Medical School, University of South China, Hengyang, China.
³ Department of Hand and Foot Surgery, The Second Affiliated Hospital, Hengyang Medical School, University of South China, Hengyang, China.

Abstract

Objective: Osteoporosis is caused by the dysregulation of bone homeostasis which is synergistically mediated by osteoclasts and osteoblasts. MiR-27a-3p is a key inhibitor of bone formation. Hence, unearthing the downstream target gene of miR-27a-3p is of great significance to understand the molecular mechanism of osteoporosis. Methods: Bioinformatics analysis was utilized to find the downstream target gene of miR-27a-3p, and dual-luciferase reporter assay was conducted to validate the interplay of miR-27a-3p and GLP1R. Besides, qRT-PCR, Western blot, and enzyme-linked immunosorbent assay (ELISA) were employed to verify the impact of miR-27a-3p on GLP1R expression and the differentiation, autophagy, and inflammatory response of MC3T3-E1 pre-osteoblasts. Results: Dual-luciferase assay validated that miR-27a-3p directly targeted GLP1R. Additionally, posttreatment of MC3T3-E1 cells with miR-27a-3p mimics resulted in a remarkable decrease in expression levels of GLP1R, cell differentiation marker gene, autophagy marker gene, and AMPK. These results indicated that miR-27a-3p targeted GLP1R to inhibit AMPK signal activation and pre-osteoblast differentiation and autophagy, while promoting the release of inflammatory factors. Conclusion: The miR-27a-3p/GLP1R regulatory axis in pre-osteoblasts contributes to understanding the molecular mechanism of osteoporosis.

Keywords: GLP1R; MiR-27a-3p; autophagy; differentiation; inflammation; osteoblast.