Cargando…
MiR-21-5p regulates extracellular matrix degradation and angiogenesis in TMJOA by targeting Spry1
BACKGROUND: Due to the lack of research on the pathological mechanism of temporomandibular joint osteoarthritis (TMJOA), there are few effective treatment measures in the clinic. In recent years, microRNAs (miRs) have been demonstrated to play an important role in the pathogenesis of osteoarthritis...
Autores principales: | , , , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2020
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7195789/ https://www.ncbi.nlm.nih.gov/pubmed/32357909 http://dx.doi.org/10.1186/s13075-020-2145-y |
_version_ | 1783528610473181184 |
---|---|
author | Ma, Shixing Zhang, Aobo Li, Xiaole Zhang, Shizhou Liu, Shaopeng Zhao, Haoming Wu, Shichao Chen, Lei Ma, Chuan Zhao, Huaqiang |
author_facet | Ma, Shixing Zhang, Aobo Li, Xiaole Zhang, Shizhou Liu, Shaopeng Zhao, Haoming Wu, Shichao Chen, Lei Ma, Chuan Zhao, Huaqiang |
author_sort | Ma, Shixing |
collection | PubMed |
description | BACKGROUND: Due to the lack of research on the pathological mechanism of temporomandibular joint osteoarthritis (TMJOA), there are few effective treatment measures in the clinic. In recent years, microRNAs (miRs) have been demonstrated to play an important role in the pathogenesis of osteoarthritis (OA) by regulating a variety of target genes, and the latest evidence shows that miR-21-5p is specifically overexpressed in OA. The purpose of this project was to clarify whether miR-21-5p can regulate the TMJOA process by targeting Spry1. METHODS: TMJOA was induced by a unilateral anterior crossbite (UAC) model, and the effect of miR-21-5p knockout on TMJOA was evaluated by toluidine blue (TB), immunohistochemical (IHC) staining, Western blotting (WB) and RT-qPCR. Primary mouse condylar chondrocytes (MCCs) were isolated, cultured and transfected with a series of mimics, inhibitors, siRNA-Spry1 or cDNA Spry1. WB, RT-qPCR, IHC and TB were used to detect the effect of miR-21-5p and its target gene Spry1 on the expression of MMP-13, VEGF and p-ERK1/2 in TMJOA. The effect of miR-21-5p on angiogenesis was evaluated by chick embryo chorioallantoic membrane (CAM) assay and WB. RESULTS: In the UAC model, the cartilage thickness and extracellular matrix of miR-21-5p knockout mice were less damaged, and miR-21-5p and UAC model were shown to affect the expression of Spry1, IL-1β, MMP-13, and VEGF. Luciferase experiments confirmed that Spry1 was the direct target of miR-21-5p. The expression levels of Spry1, MMP-13, VEGF and p-ERK1/2 in MCCs transfected with miR-21-5p mimic were higher than those in the inhibitor group. Under the simulated inflammatory environment of IL-1β, the expression levels of MMP-13, VEGF and p-ERK1/2 were positively correlated with miR-21-5p, while Spry1 was negatively correlated with miR-21-5p. Inhibition of miR-21-5p expression and overexpression of Spry1 enhanced the inhibition of MMP-13, VEGF and p-ERK1/2 expression. MiR-21-5p had a significant role in promoting angiogenesis in the chick embryo CAM assay, and this role was clearly mediated by the ERK-MAPK signalling pathway. CONCLUSION: This study verified that miR-21-5p can promote the process of TMJOA by targeting Spry1, which provides a new direction for future research on the treatment of this disease. |
format | Online Article Text |
id | pubmed-7195789 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-71957892020-05-06 MiR-21-5p regulates extracellular matrix degradation and angiogenesis in TMJOA by targeting Spry1 Ma, Shixing Zhang, Aobo Li, Xiaole Zhang, Shizhou Liu, Shaopeng Zhao, Haoming Wu, Shichao Chen, Lei Ma, Chuan Zhao, Huaqiang Arthritis Res Ther Research Article BACKGROUND: Due to the lack of research on the pathological mechanism of temporomandibular joint osteoarthritis (TMJOA), there are few effective treatment measures in the clinic. In recent years, microRNAs (miRs) have been demonstrated to play an important role in the pathogenesis of osteoarthritis (OA) by regulating a variety of target genes, and the latest evidence shows that miR-21-5p is specifically overexpressed in OA. The purpose of this project was to clarify whether miR-21-5p can regulate the TMJOA process by targeting Spry1. METHODS: TMJOA was induced by a unilateral anterior crossbite (UAC) model, and the effect of miR-21-5p knockout on TMJOA was evaluated by toluidine blue (TB), immunohistochemical (IHC) staining, Western blotting (WB) and RT-qPCR. Primary mouse condylar chondrocytes (MCCs) were isolated, cultured and transfected with a series of mimics, inhibitors, siRNA-Spry1 or cDNA Spry1. WB, RT-qPCR, IHC and TB were used to detect the effect of miR-21-5p and its target gene Spry1 on the expression of MMP-13, VEGF and p-ERK1/2 in TMJOA. The effect of miR-21-5p on angiogenesis was evaluated by chick embryo chorioallantoic membrane (CAM) assay and WB. RESULTS: In the UAC model, the cartilage thickness and extracellular matrix of miR-21-5p knockout mice were less damaged, and miR-21-5p and UAC model were shown to affect the expression of Spry1, IL-1β, MMP-13, and VEGF. Luciferase experiments confirmed that Spry1 was the direct target of miR-21-5p. The expression levels of Spry1, MMP-13, VEGF and p-ERK1/2 in MCCs transfected with miR-21-5p mimic were higher than those in the inhibitor group. Under the simulated inflammatory environment of IL-1β, the expression levels of MMP-13, VEGF and p-ERK1/2 were positively correlated with miR-21-5p, while Spry1 was negatively correlated with miR-21-5p. Inhibition of miR-21-5p expression and overexpression of Spry1 enhanced the inhibition of MMP-13, VEGF and p-ERK1/2 expression. MiR-21-5p had a significant role in promoting angiogenesis in the chick embryo CAM assay, and this role was clearly mediated by the ERK-MAPK signalling pathway. CONCLUSION: This study verified that miR-21-5p can promote the process of TMJOA by targeting Spry1, which provides a new direction for future research on the treatment of this disease. BioMed Central 2020-05-01 2020 /pmc/articles/PMC7195789/ /pubmed/32357909 http://dx.doi.org/10.1186/s13075-020-2145-y Text en © The Author(s). 2020 Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data. |
spellingShingle | Research Article Ma, Shixing Zhang, Aobo Li, Xiaole Zhang, Shizhou Liu, Shaopeng Zhao, Haoming Wu, Shichao Chen, Lei Ma, Chuan Zhao, Huaqiang MiR-21-5p regulates extracellular matrix degradation and angiogenesis in TMJOA by targeting Spry1 |
title | MiR-21-5p regulates extracellular matrix degradation and angiogenesis in TMJOA by targeting Spry1 |
title_full | MiR-21-5p regulates extracellular matrix degradation and angiogenesis in TMJOA by targeting Spry1 |
title_fullStr | MiR-21-5p regulates extracellular matrix degradation and angiogenesis in TMJOA by targeting Spry1 |
title_full_unstemmed | MiR-21-5p regulates extracellular matrix degradation and angiogenesis in TMJOA by targeting Spry1 |
title_short | MiR-21-5p regulates extracellular matrix degradation and angiogenesis in TMJOA by targeting Spry1 |
title_sort | mir-21-5p regulates extracellular matrix degradation and angiogenesis in tmjoa by targeting spry1 |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7195789/ https://www.ncbi.nlm.nih.gov/pubmed/32357909 http://dx.doi.org/10.1186/s13075-020-2145-y |
work_keys_str_mv | AT mashixing mir215pregulatesextracellularmatrixdegradationandangiogenesisintmjoabytargetingspry1 AT zhangaobo mir215pregulatesextracellularmatrixdegradationandangiogenesisintmjoabytargetingspry1 AT lixiaole mir215pregulatesextracellularmatrixdegradationandangiogenesisintmjoabytargetingspry1 AT zhangshizhou mir215pregulatesextracellularmatrixdegradationandangiogenesisintmjoabytargetingspry1 AT liushaopeng mir215pregulatesextracellularmatrixdegradationandangiogenesisintmjoabytargetingspry1 AT zhaohaoming mir215pregulatesextracellularmatrixdegradationandangiogenesisintmjoabytargetingspry1 AT wushichao mir215pregulatesextracellularmatrixdegradationandangiogenesisintmjoabytargetingspry1 AT chenlei mir215pregulatesextracellularmatrixdegradationandangiogenesisintmjoabytargetingspry1 AT machuan mir215pregulatesextracellularmatrixdegradationandangiogenesisintmjoabytargetingspry1 AT zhaohuaqiang mir215pregulatesextracellularmatrixdegradationandangiogenesisintmjoabytargetingspry1 |