The Inhibition of Osteoblast Viability by Monosodium Urate Crystal-Stimulated Neutrophil-Derived Exosomes
Background and ObjectiveBone erosion is common in patients with gout. The role of neutrophil-derived exosomes in gouty bone erosion remains elusive. This study aimed to investigate the functions of the neutrophil-derived exosomes in the development of bone erosion in gout.MethodsNeutrophil-derived e...
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Frontiers Media S.A.
2022-05-01
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| Series: | Frontiers in Immunology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fimmu.2022.809586/full |
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| author | Ertao Jia Ertao Jia Haiqiong Zhu Hongling Geng Li Zhong Li Zhong Xia Qiu Xia Qiu Jingjing Xie Jingjing Xie Yuya Xiao Yuya Xiao Yubao Jiang Yubao Jiang Min Xiao Min Xiao Yanying Zhang Yanying Zhang Jiaxin Wei Jiaxin Wei Dabin Tang Dabin Tang Jianyong Zhang Jianyong Zhang |
| author_facet | Ertao Jia Ertao Jia Haiqiong Zhu Hongling Geng Li Zhong Li Zhong Xia Qiu Xia Qiu Jingjing Xie Jingjing Xie Yuya Xiao Yuya Xiao Yubao Jiang Yubao Jiang Min Xiao Min Xiao Yanying Zhang Yanying Zhang Jiaxin Wei Jiaxin Wei Dabin Tang Dabin Tang Jianyong Zhang Jianyong Zhang |
| author_sort | Ertao Jia |
| collection | DOAJ |
| description | Background and ObjectiveBone erosion is common in patients with gout. The role of neutrophil-derived exosomes in gouty bone erosion remains elusive. This study aimed to investigate the functions of the neutrophil-derived exosomes in the development of bone erosion in gout.MethodsNeutrophil-derived exosomes were collected and assessed by transmission electron microscopy and nanoparticle tracking analysis. Cell counting kit-8 assay was applied to evaluate cell viability, and cell apoptosis was assessed by flow cytometry. In addition, quantitative Real-time PCR and Western blotting were used to determine the expression levels of alkaline phosphatase (ALP), osteoprotegerin (OPG), and receptor activator of nuclear factor-κB ligand (RANKL). Neutrophil-derived exosomes were tagged with PKH67. The miRNA expression profiles of exosomes and human fetal osteoblasts (hFOB) were compared using high-throughput sequencing. Functional miRNAs transfected into hFOB after co-incubation with exosomes were selected and validated by preliminary qPCR.ResultsNeutrophil-derived exosomes were stimulated by monosodium urate (MSU). The exosomes could inhibit the viability of the hFOB, and the expression levels of ALP and OPG were down-regulated, while the expression level of RANKL was up-regulated. However, there was no significant difference in the viability of osteoclasts and the expression of nuclear factor of activated T cells 1. Exosomes were observed in the cytoplasm under a confocal microscopy, confirming that exosomes could be taken up by hFOB. In total, 2590 miRNAs were found, of which 47 miRNAs were differentially expressed. Among the delivered miRNAs, miR-1246 exhibited the highest level of differential expression. The viability of hFOB was reduced by miR-1246 mimics and increased by miR-1246 inhibitors. There was no significant difference in hFOB apoptosis rate between the miR-1246 mimic and miR-1246 inhibitor group. MiR-1246 overexpression decreased the expression levels of ALP and OPG, whereas increasing the expression level of RANKL. In contrast, miR-1246 inhibitor increased the expression levels of ALP and OPG, while decreasing the expression level of RANKL. Neutrophil-derived exosomes stimulated by MSU could increase the expression of miR-1246. ConclusionNeutrophil-derived exosomes stimulated by MSU could inhibit the viability of osteoblasts. |
| first_indexed | 2024-04-12T11:48:43Z |
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| language | English |
| last_indexed | 2024-04-12T11:48:43Z |
| publishDate | 2022-05-01 |
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| series | Frontiers in Immunology |
| spelling | doaj.art-c82a78d91943492687e455f36019f4872022-12-22T03:34:15ZengFrontiers Media S.A.Frontiers in Immunology1664-32242022-05-011310.3389/fimmu.2022.809586809586The Inhibition of Osteoblast Viability by Monosodium Urate Crystal-Stimulated Neutrophil-Derived ExosomesErtao Jia0Ertao Jia1Haiqiong Zhu2Hongling Geng3Li Zhong4Li Zhong5Xia Qiu6Xia Qiu7Jingjing Xie8Jingjing Xie9Yuya Xiao10Yuya Xiao11Yubao Jiang12Yubao Jiang13Min Xiao14Min Xiao15Yanying Zhang16Yanying Zhang17Jiaxin Wei18Jiaxin Wei19Dabin Tang20Dabin Tang21Jianyong Zhang22Jianyong Zhang23The Department of Rheumatology, Shenzhen Traditional Chinese Medicine Hospital, Shenzhen, ChinaThe Department of Rheumatology, The Fourth Clinical Medical College of Guangzhou University of Chinese Medicine, Shenzhen, ChinaShenzhen Traditional Chinese Medicine Hospital Affiliated to Nanjing University of Chinese Medicine, Shenzhen, ChinaThe Department of Gynecology, Guangdong Provincial Hospital of Chinese Medicine, The Second Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou, ChinaThe Department of Rheumatology, Shenzhen Traditional Chinese Medicine Hospital, Shenzhen, ChinaThe Department of Rheumatology, The Fourth Clinical Medical College of Guangzhou University of Chinese Medicine, Shenzhen, ChinaThe Department of Rheumatology, Shenzhen Traditional Chinese Medicine Hospital, Shenzhen, ChinaThe Department of Rheumatology, The Fourth Clinical Medical College of Guangzhou University of Chinese Medicine, Shenzhen, ChinaThe Department of Rheumatology, Shenzhen Traditional Chinese Medicine Hospital, Shenzhen, ChinaThe Department of Rheumatology, The Fourth Clinical Medical College of Guangzhou University of Chinese Medicine, Shenzhen, ChinaThe Department of Rheumatology, Shenzhen Traditional Chinese Medicine Hospital, Shenzhen, ChinaThe Department of Rheumatology, The Fourth Clinical Medical College of Guangzhou University of Chinese Medicine, Shenzhen, ChinaThe Department of Rheumatology, Shenzhen Traditional Chinese Medicine Hospital, Shenzhen, ChinaThe Department of Rheumatology, The Fourth Clinical Medical College of Guangzhou University of Chinese Medicine, Shenzhen, ChinaThe Department of Rheumatology, Shenzhen Traditional Chinese Medicine Hospital, Shenzhen, ChinaThe Department of Rheumatology, The Fourth Clinical Medical College of Guangzhou University of Chinese Medicine, Shenzhen, ChinaThe Department of Rheumatology, Shenzhen Traditional Chinese Medicine Hospital, Shenzhen, ChinaThe Department of Rheumatology, The Fourth Clinical Medical College of Guangzhou University of Chinese Medicine, Shenzhen, ChinaThe Department of Rheumatology, Shenzhen Traditional Chinese Medicine Hospital, Shenzhen, ChinaThe Department of Rheumatology, The Fourth Clinical Medical College of Guangzhou University of Chinese Medicine, Shenzhen, ChinaThe Department of Rheumatology, Shenzhen Traditional Chinese Medicine Hospital, Shenzhen, ChinaThe Department of Rheumatology, The Fourth Clinical Medical College of Guangzhou University of Chinese Medicine, Shenzhen, ChinaThe Department of Rheumatology, Shenzhen Traditional Chinese Medicine Hospital, Shenzhen, ChinaThe Department of Rheumatology, The Fourth Clinical Medical College of Guangzhou University of Chinese Medicine, Shenzhen, ChinaBackground and ObjectiveBone erosion is common in patients with gout. The role of neutrophil-derived exosomes in gouty bone erosion remains elusive. This study aimed to investigate the functions of the neutrophil-derived exosomes in the development of bone erosion in gout.MethodsNeutrophil-derived exosomes were collected and assessed by transmission electron microscopy and nanoparticle tracking analysis. Cell counting kit-8 assay was applied to evaluate cell viability, and cell apoptosis was assessed by flow cytometry. In addition, quantitative Real-time PCR and Western blotting were used to determine the expression levels of alkaline phosphatase (ALP), osteoprotegerin (OPG), and receptor activator of nuclear factor-κB ligand (RANKL). Neutrophil-derived exosomes were tagged with PKH67. The miRNA expression profiles of exosomes and human fetal osteoblasts (hFOB) were compared using high-throughput sequencing. Functional miRNAs transfected into hFOB after co-incubation with exosomes were selected and validated by preliminary qPCR.ResultsNeutrophil-derived exosomes were stimulated by monosodium urate (MSU). The exosomes could inhibit the viability of the hFOB, and the expression levels of ALP and OPG were down-regulated, while the expression level of RANKL was up-regulated. However, there was no significant difference in the viability of osteoclasts and the expression of nuclear factor of activated T cells 1. Exosomes were observed in the cytoplasm under a confocal microscopy, confirming that exosomes could be taken up by hFOB. In total, 2590 miRNAs were found, of which 47 miRNAs were differentially expressed. Among the delivered miRNAs, miR-1246 exhibited the highest level of differential expression. The viability of hFOB was reduced by miR-1246 mimics and increased by miR-1246 inhibitors. There was no significant difference in hFOB apoptosis rate between the miR-1246 mimic and miR-1246 inhibitor group. MiR-1246 overexpression decreased the expression levels of ALP and OPG, whereas increasing the expression level of RANKL. In contrast, miR-1246 inhibitor increased the expression levels of ALP and OPG, while decreasing the expression level of RANKL. Neutrophil-derived exosomes stimulated by MSU could increase the expression of miR-1246. ConclusionNeutrophil-derived exosomes stimulated by MSU could inhibit the viability of osteoblasts.https://www.frontiersin.org/articles/10.3389/fimmu.2022.809586/fullgoutneutrophil-derived exosomesosteoclastsosteoblastsmonosodium urate crystals |
| spellingShingle | Ertao Jia Ertao Jia Haiqiong Zhu Hongling Geng Li Zhong Li Zhong Xia Qiu Xia Qiu Jingjing Xie Jingjing Xie Yuya Xiao Yuya Xiao Yubao Jiang Yubao Jiang Min Xiao Min Xiao Yanying Zhang Yanying Zhang Jiaxin Wei Jiaxin Wei Dabin Tang Dabin Tang Jianyong Zhang Jianyong Zhang The Inhibition of Osteoblast Viability by Monosodium Urate Crystal-Stimulated Neutrophil-Derived Exosomes Frontiers in Immunology gout neutrophil-derived exosomes osteoclasts osteoblasts monosodium urate crystals |
| title | The Inhibition of Osteoblast Viability by Monosodium Urate Crystal-Stimulated Neutrophil-Derived Exosomes |
| title_full | The Inhibition of Osteoblast Viability by Monosodium Urate Crystal-Stimulated Neutrophil-Derived Exosomes |
| title_fullStr | The Inhibition of Osteoblast Viability by Monosodium Urate Crystal-Stimulated Neutrophil-Derived Exosomes |
| title_full_unstemmed | The Inhibition of Osteoblast Viability by Monosodium Urate Crystal-Stimulated Neutrophil-Derived Exosomes |
| title_short | The Inhibition of Osteoblast Viability by Monosodium Urate Crystal-Stimulated Neutrophil-Derived Exosomes |
| title_sort | inhibition of osteoblast viability by monosodium urate crystal stimulated neutrophil derived exosomes |
| topic | gout neutrophil-derived exosomes osteoclasts osteoblasts monosodium urate crystals |
| url | https://www.frontiersin.org/articles/10.3389/fimmu.2022.809586/full |
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