Diabetic Macrophage Exosomal miR-381-3p Inhibits Epithelial Cell Autophagy Via NR5A2

Xin Huang; Linhesheng Wei; Mengdi Li; Yong Zhang; Shuhong Kuang; Zongshan Shen; Hui Liu; Zhengmei Lin

doi:10.1016/j.identj.2024.02.001

Diabetic Macrophage Exosomal miR-381-3p Inhibits Epithelial Cell Autophagy Via NR5A2

Int Dent J. 2024 Apr 28:S0020-6539(24)00055-8. doi: 10.1016/j.identj.2024.02.001. Online ahead of print.

Authors

Xin Huang¹, Linhesheng Wei¹, Mengdi Li², Yong Zhang¹, Shuhong Kuang¹, Zongshan Shen¹, Hui Liu¹, Zhengmei Lin³

Affiliations

¹ Hospital of Stomatology, Guangdong Provincial Key Laboratory of Stomatology, Guanghua School of Stomatology, Sun Yat-sen University, Guangzhou, Guangdong, China.
² Department of Periodontology, 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, China.
³ Hospital of Stomatology, Guangdong Provincial Key Laboratory of Stomatology, Guanghua School of Stomatology, Sun Yat-sen University, Guangzhou, Guangdong, China. Electronic address: linzhm@mail.sysu.edu.cn.

PMID: 38685137
DOI: 10.1016/j.identj.2024.02.001

Abstract

Purpose: To explore the mechanism underlying autophagy disruption in gingival epithelial cells (GECs) in diabetic individuals.

Methods and materials: Bone marrow-derived macrophages (BMDMs) and GECs were extracted from C57/bl and db/db mice, the exosomes (Exo) were isolated from BMDMs. qRT‒PCR and Western blotting were performed to analyse gene expression. The AnimalTFDB database was used to identify relevant transcription factors, and miRNA sequencing was utilised to identify relevant miRNAs with the aid of the TargetScan/miRDB/miRWalk databases. A dual-luciferase assay was conducted to verify intermolecular targeting relationships.

Results: Similar to BMDMs, BMDM-derived Exos disrupted autophagy and exerted proinflammatory effects in GEC cocultures, and ATG7 may play a vital role. AnimalTFDB database analysis and dual-luciferase assays indicated that NR5A2 is the most relevant transcription factor that regulates Atg7 expression. SiRNA-NR5A2 transfection blocked autophagy in GECs and exacerbated inflammation, whereas NR5A2 upregulation restored ATG7 expression and ameliorated Exo^DM-mediated inflammation. MiRNA sequencing, with TargetScan/miRDB/miRWalk analyses and dual-luciferase assays, confirmed that miR-381-3p is the most relevant miRNA that targets NR5A2. MiR-381-3p mimic transfection blocked autophagy in GECs and exacerbated inflammation, while miR-381-3p inhibitor transfection restored ATG7 expression and attenuated Exo^DM-mediated inflammation.

Conclusion: BMDM-derived Exos, which carry miR-381-3p, inhibit NR5A2 and disrupt autophagy in GECs, increasing periodontal inflammation in diabetes.

Keywords: Autophagy Flux; Diabetic Periodontitis; Macrophage Epithelial Cell Interaction; NR5A2; miR-381-3p.