MZB1 targeted by miR‐185‐5p inhibits the migration of human periodontal ligament cells through NF‐κB signaling and promotes alveolar bone loss

Abstract

Objective To explore the role of Marginal Zone B and B‐1 Cell‐Specific Protein (MZB1), a novel molecule associated with periodontitis, in migration of human periodontal ligament cells (hPDLCs) and alveolar bone orchestration. Background MZB1 is an ER‐localized protein and its upregulation has been found to be associated with a variety of human diseases. However, few studies have investigated the effect and mechanism of MZB1 on hPDLCs in periodontitis. Methods Gene expression profiles in human gingival tissues were acquired from the Gene Expression Omnibus (GEO) database, and candidate molecules were then selected through bioinformatic analysis. Subsequently, we identified the localization and expression of MZB1 in human gingival tissues, mice, and hPDLCs by immunofluorescence, RT‐qPCR, and Western blot. Dual‐luciferase reporter assay was applied to assess the binding of miR‐185‐5p to MZB1. Furthermore, the effects of MZB1 on cell migration, proliferation, and apoptosis in vitro were investigated by wound‐healing assay, transwell assay, CCK‐8 assay, and flow cytometry analysis. Finally, Micro‐CT analysis and H&E staining were performed to examine the effects of MZB1 on alveolar bone loss in vivo. Results Bioinformatic analysis discovered that MZB1 was one of the most significantly increased genes in periodontitis patients. MZB1 was markedly increased in the gingival tissues of periodontitis patients, in the mouse models, and in the hPDLCs treated with lipopolysaccharide of Porphyromonas gingivalis (LPS‐PG). Furthermore, in vitro experiments showed that MZB1, as a target gene of miR‐185‐5p, inhibited migration of hPDLCs. Overexpression of MZB1 specifically upregulated the phosphorylation of p65, while pretreatment of MZB1‐overexpressed hPDLCs with PDTC (NF‐κB inhibitor) notably reduced the p‐p65 level and promoted cell migration. In addition, the mRNA expression levels of alkaline phosphatase (ALP) and Runt‐related transcription factor 2 (Runx2) were inhibited in MZB1‐overexpressed hPDLCs and miR‐185‐5p inhibitor treated hPDLCs, respectively. In vivo experiments showed that knockdown of MZB1 alleviated the loss of alveolar bone. Conclusion As a target gene of miR‐185‐5p, MZB1 plays a crucial role in inhibiting the migration of hPDLCs through NF‐κB signaling pathway and deteriorating alveolar bone loss.

Comprehensive analysis of circRNA expression profiles in p75NTR-knockout ecto- mesenchymal stem cells

“…Based on the prediction of circRNA-miRNA, we selected three circRNAs as potential fundamental molecules in the circRNA-miRNA-mRNA network.it is reported that the miR-185-5p inhibits mRNA expression levels of Runx2 in hPDLCs (Li et al 2022). This nding was accordant with the circRNA-miRNA-mRNA network(Fig.…”

Altered expression of MZB1 in periodontitis: A possible link to disease pathogenesis

“…This result is consistent with the previous studies that have demonstrated the upregulation of MZB1 in patients with aggressive periodontitis and chronic periodontitis. 2,3,8,[21][22][23] The MZB1 gene (marginal zone B-and B1-cell-specific protein) encodes for the protein MZB1 and is defined in the human gene database 24 as a gene that interacts with both heavy and light chains of IgM (immunoglobulin M) to facilitate its assembly and secretion. The pathways related to the MZB1 gene encompass positive regulation of the biosynthesis process of immunoglobulin, apoptosis, cell proliferation, B-cell proliferation regulation, and integrin activation.…”

Recent Clinical Treatment and Basic Research on the Alveolar Bone

“…However, a few studies have examined the effect of MZB1 on human periodontal ligament cells (hPDLCs) in the presence of periodontitis and the mechanisms underlying this effect. Li et al performed experiments to determine the role of hPDLCs in migration and alveolar bone organization [ 148 ]. Bioinformatics analysis revealed that MZB1 is one of the most significantly upregulated genes in patients with periodontitis.…”

“…Bioinformatics analysis revealed that MZB1 is one of the most significantly upregulated genes in patients with periodontitis. mZB1, as a target gene of miR-185-5p, plays an important role in suppressing hPDLC migration through the NF-κB signaling pathway and exacerbating alveolar bone loss [ 148 ]. Bioinformatics analysis was used in this study [ 148 ].…”

“…mZB1, as a target gene of miR-185-5p, plays an important role in suppressing hPDLC migration through the NF-κB signaling pathway and exacerbating alveolar bone loss [ 148 ]. Bioinformatics analysis was used in this study [ 148 ]. This analytical method has attracted much attention in oral research, and with this analysis, various mechanisms of alveolar bone will be revealed in the future.…”