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MicroRNA-200b relieves LPS-induced inflammatory injury by targeting FUT4 in knee articular chondrocytes in vitro
Osteoarthritis (OA), characterized by the degeneration of articular cartilage, is a major problem in aging populations, and cartilage chondrocytes have been indicated to serve a curial role in the progression of OA. MicroRNA-200b-3p (miR-200b) was preliminarily identified to participate in OA. Howev...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
D.A. Spandidos
2021
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7938448/ https://www.ncbi.nlm.nih.gov/pubmed/33692838 http://dx.doi.org/10.3892/etm.2021.9838 |
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author | Li, Yintai Wei, Suizhuan Zhang, Zhongping |
author_facet | Li, Yintai Wei, Suizhuan Zhang, Zhongping |
author_sort | Li, Yintai |
collection | PubMed |
description | Osteoarthritis (OA), characterized by the degeneration of articular cartilage, is a major problem in aging populations, and cartilage chondrocytes have been indicated to serve a curial role in the progression of OA. MicroRNA-200b-3p (miR-200b) was preliminarily identified to participate in OA. However, its role and mechanism of action in injured chondrocytes in OA remain unclear to date. In the present study, lipopolysaccharide (LPS)-treated cells isolated from normal knee articular cartilage were used to mimic inflammatory injury of OA chondrocytes. Cell viability, apoptosis and inflammatory responses were detected using Cell Counting Kit-8, flow cytometry and enzyme-linked immunosorbent assay, respectively. The expression levels of miR-200b and fucosyltransferase-4 (FUT4) were measured by reverse transcription-quantitative PCR and western blotting. The association between miR-200b and FUT4 was verified using TargetScan software, dual-luciferase reporter assay and RNA immunoprecipitation. The results indicated that LPS treatment decreased cell viability of primary chondrocytes, and increased apoptosis rate and production of IL-1β, IL-6 and TNF-α. The expression level of miR-200b was downregulated, and that of FUT4 was upregulated in OA cartilage tissues and LPS-treated normal chondrocytes compared with normal cartilage tissues and chondrocytes. Overexpression of miR-200b via transfection with miR-200b mimic inhibited the apoptosis rate and reduced the levels of IL-1β, IL-6 and TNF-α in LPS-stimulated chondrocytes. However, the suppressive effect of miR-200b overexpression on the LPS-induced inflammatory injury in chondrocytes was reversed by the restoration of FUT4 levels. Notably, FUT4 was indicated to be a downstream target of miR-200b and was negatively regulated by miR-200b. Taken together, the results of the current study indicated that miR-200b protected chondrocytes from LPS-induced inflammatory injury in vitro by targeting FUT4. These findings revealed the miR-200b/FUT4 axis as a potential candidate to target the degeneration of cartilages, thereby inhibiting the progression of OA. |
format | Online Article Text |
id | pubmed-7938448 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | D.A. Spandidos |
record_format | MEDLINE/PubMed |
spelling | pubmed-79384482021-03-09 MicroRNA-200b relieves LPS-induced inflammatory injury by targeting FUT4 in knee articular chondrocytes in vitro Li, Yintai Wei, Suizhuan Zhang, Zhongping Exp Ther Med Articles Osteoarthritis (OA), characterized by the degeneration of articular cartilage, is a major problem in aging populations, and cartilage chondrocytes have been indicated to serve a curial role in the progression of OA. MicroRNA-200b-3p (miR-200b) was preliminarily identified to participate in OA. However, its role and mechanism of action in injured chondrocytes in OA remain unclear to date. In the present study, lipopolysaccharide (LPS)-treated cells isolated from normal knee articular cartilage were used to mimic inflammatory injury of OA chondrocytes. Cell viability, apoptosis and inflammatory responses were detected using Cell Counting Kit-8, flow cytometry and enzyme-linked immunosorbent assay, respectively. The expression levels of miR-200b and fucosyltransferase-4 (FUT4) were measured by reverse transcription-quantitative PCR and western blotting. The association between miR-200b and FUT4 was verified using TargetScan software, dual-luciferase reporter assay and RNA immunoprecipitation. The results indicated that LPS treatment decreased cell viability of primary chondrocytes, and increased apoptosis rate and production of IL-1β, IL-6 and TNF-α. The expression level of miR-200b was downregulated, and that of FUT4 was upregulated in OA cartilage tissues and LPS-treated normal chondrocytes compared with normal cartilage tissues and chondrocytes. Overexpression of miR-200b via transfection with miR-200b mimic inhibited the apoptosis rate and reduced the levels of IL-1β, IL-6 and TNF-α in LPS-stimulated chondrocytes. However, the suppressive effect of miR-200b overexpression on the LPS-induced inflammatory injury in chondrocytes was reversed by the restoration of FUT4 levels. Notably, FUT4 was indicated to be a downstream target of miR-200b and was negatively regulated by miR-200b. Taken together, the results of the current study indicated that miR-200b protected chondrocytes from LPS-induced inflammatory injury in vitro by targeting FUT4. These findings revealed the miR-200b/FUT4 axis as a potential candidate to target the degeneration of cartilages, thereby inhibiting the progression of OA. D.A. Spandidos 2021-04 2021-02-25 /pmc/articles/PMC7938448/ /pubmed/33692838 http://dx.doi.org/10.3892/etm.2021.9838 Text en Copyright: © Li et al. This is an open access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs License (https://creativecommons.org/licenses/by-nc-nd/4.0/) , which permits use and distribution in any medium, provided the original work is properly cited, the use is non-commercial and no modifications or adaptations are made. |
spellingShingle | Articles Li, Yintai Wei, Suizhuan Zhang, Zhongping MicroRNA-200b relieves LPS-induced inflammatory injury by targeting FUT4 in knee articular chondrocytes in vitro |
title | MicroRNA-200b relieves LPS-induced inflammatory injury by targeting FUT4 in knee articular chondrocytes in vitro |
title_full | MicroRNA-200b relieves LPS-induced inflammatory injury by targeting FUT4 in knee articular chondrocytes in vitro |
title_fullStr | MicroRNA-200b relieves LPS-induced inflammatory injury by targeting FUT4 in knee articular chondrocytes in vitro |
title_full_unstemmed | MicroRNA-200b relieves LPS-induced inflammatory injury by targeting FUT4 in knee articular chondrocytes in vitro |
title_short | MicroRNA-200b relieves LPS-induced inflammatory injury by targeting FUT4 in knee articular chondrocytes in vitro |
title_sort | microrna-200b relieves lps-induced inflammatory injury by targeting fut4 in knee articular chondrocytes in vitro |
topic | Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7938448/ https://www.ncbi.nlm.nih.gov/pubmed/33692838 http://dx.doi.org/10.3892/etm.2021.9838 |
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