Cargando…

FTY-720P Suppresses Osteoclast Formation by Regulating Expression of Interleukin-6 (IL-6), Interleukin-4 (IL-4), and Matrix Metalloproteinase 2 (MMP-2)

BACKGROUND: Osteoclast formation is closely related to the immune system. FTY720, a new immunosuppressive agent, has some functions in immune regulation. Its main active ingredients become FTY-720P in vivo by phosphorylation modification. The objective of this study was to determine the effects of F...

Descripción completa

Detalles Bibliográficos
Autores principales: Zhang, Dawei, Huang, Yongjun, Huang, Zongwen, Zhang, Rongkai, Wang, Honggang, Huang, Dong
Formato: Online Artículo Texto
Lenguaje:English
Publicado: International Scientific Literature, Inc. 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4924886/
https://www.ncbi.nlm.nih.gov/pubmed/27344392
http://dx.doi.org/10.12659/MSM.896690
_version_ 1782439944628731904
author Zhang, Dawei
Huang, Yongjun
Huang, Zongwen
Zhang, Rongkai
Wang, Honggang
Huang, Dong
author_facet Zhang, Dawei
Huang, Yongjun
Huang, Zongwen
Zhang, Rongkai
Wang, Honggang
Huang, Dong
author_sort Zhang, Dawei
collection PubMed
description BACKGROUND: Osteoclast formation is closely related to the immune system. FTY720, a new immunosuppressive agent, has some functions in immune regulation. Its main active ingredients become FTY-720P in vivo by phosphorylation modification. The objective of this study was to determine the effects of FTY-720 with various concentrations on osteoclasts in vitro. MATERIAL/METHODS: RAW264.7 cells and bone marrow-derived mononuclear phagocytes (BMMs) were treated with RANKL to obtain osteoclasts in vitro. To investigate the role of FTY-720 in osteoclast formation, trap enzyme staining was performed and the number of osteoclasts was counted. Bone slices were stained with methylene blue, we counted the number of lacunae after bone slices were placed into dishes together with osteoclasts, and we observed the effect and function of FTY-720 in osteoclasts induced by RAW264.7 cells and BMMs. Then, we used a protein array kit to explore the effects of FTY-720P on osteoclasts. RESULTS: The results of enzyme trap staining and F-actin staining experiments show that, with the increasing concentration of FTY-720P, the number of osteoclast induced by RAW264.7 cells and BMMs gradually decreased (P<0.05), especially when the FTY-720P concentration reached 1000 ng/ml, and the number of osteoclasts formed was the lowest (P<0.05). With bone lacuna toluidine blue staining, the results also show that, with the increasing concentration of FTY-720P, the number of bone lacuna gradually decreased (P<0.05), and the number of lacunae is lowest when the concentration reached 800 ng/ml. Finally, protein array results showed that IL-4, IL-6, IL-12, MMP-2, VEGF-C, GFR, basic FGF, MIP-2, and insulin proteins were regulated after FTY-720P treatment. CONCLUSIONS: FTY-720P can suppress osteoclast formation and function, and FTY-720P induces a series of cytokine changes.
format Online
Article
Text
id pubmed-4924886
institution National Center for Biotechnology Information
language English
publishDate 2016
publisher International Scientific Literature, Inc.
record_format MEDLINE/PubMed
spelling pubmed-49248862016-07-11 FTY-720P Suppresses Osteoclast Formation by Regulating Expression of Interleukin-6 (IL-6), Interleukin-4 (IL-4), and Matrix Metalloproteinase 2 (MMP-2) Zhang, Dawei Huang, Yongjun Huang, Zongwen Zhang, Rongkai Wang, Honggang Huang, Dong Med Sci Monit Lab/In Vitro Research BACKGROUND: Osteoclast formation is closely related to the immune system. FTY720, a new immunosuppressive agent, has some functions in immune regulation. Its main active ingredients become FTY-720P in vivo by phosphorylation modification. The objective of this study was to determine the effects of FTY-720 with various concentrations on osteoclasts in vitro. MATERIAL/METHODS: RAW264.7 cells and bone marrow-derived mononuclear phagocytes (BMMs) were treated with RANKL to obtain osteoclasts in vitro. To investigate the role of FTY-720 in osteoclast formation, trap enzyme staining was performed and the number of osteoclasts was counted. Bone slices were stained with methylene blue, we counted the number of lacunae after bone slices were placed into dishes together with osteoclasts, and we observed the effect and function of FTY-720 in osteoclasts induced by RAW264.7 cells and BMMs. Then, we used a protein array kit to explore the effects of FTY-720P on osteoclasts. RESULTS: The results of enzyme trap staining and F-actin staining experiments show that, with the increasing concentration of FTY-720P, the number of osteoclast induced by RAW264.7 cells and BMMs gradually decreased (P<0.05), especially when the FTY-720P concentration reached 1000 ng/ml, and the number of osteoclasts formed was the lowest (P<0.05). With bone lacuna toluidine blue staining, the results also show that, with the increasing concentration of FTY-720P, the number of bone lacuna gradually decreased (P<0.05), and the number of lacunae is lowest when the concentration reached 800 ng/ml. Finally, protein array results showed that IL-4, IL-6, IL-12, MMP-2, VEGF-C, GFR, basic FGF, MIP-2, and insulin proteins were regulated after FTY-720P treatment. CONCLUSIONS: FTY-720P can suppress osteoclast formation and function, and FTY-720P induces a series of cytokine changes. International Scientific Literature, Inc. 2016-06-26 /pmc/articles/PMC4924886/ /pubmed/27344392 http://dx.doi.org/10.12659/MSM.896690 Text en © Med Sci Monit, 2016 This work is licensed under Creative Common Attribution-NonCommercial-NoDerivatives 4.0 International (CC BY-NC-ND 4.0)
spellingShingle Lab/In Vitro Research
Zhang, Dawei
Huang, Yongjun
Huang, Zongwen
Zhang, Rongkai
Wang, Honggang
Huang, Dong
FTY-720P Suppresses Osteoclast Formation by Regulating Expression of Interleukin-6 (IL-6), Interleukin-4 (IL-4), and Matrix Metalloproteinase 2 (MMP-2)
title FTY-720P Suppresses Osteoclast Formation by Regulating Expression of Interleukin-6 (IL-6), Interleukin-4 (IL-4), and Matrix Metalloproteinase 2 (MMP-2)
title_full FTY-720P Suppresses Osteoclast Formation by Regulating Expression of Interleukin-6 (IL-6), Interleukin-4 (IL-4), and Matrix Metalloproteinase 2 (MMP-2)
title_fullStr FTY-720P Suppresses Osteoclast Formation by Regulating Expression of Interleukin-6 (IL-6), Interleukin-4 (IL-4), and Matrix Metalloproteinase 2 (MMP-2)
title_full_unstemmed FTY-720P Suppresses Osteoclast Formation by Regulating Expression of Interleukin-6 (IL-6), Interleukin-4 (IL-4), and Matrix Metalloproteinase 2 (MMP-2)
title_short FTY-720P Suppresses Osteoclast Formation by Regulating Expression of Interleukin-6 (IL-6), Interleukin-4 (IL-4), and Matrix Metalloproteinase 2 (MMP-2)
title_sort fty-720p suppresses osteoclast formation by regulating expression of interleukin-6 (il-6), interleukin-4 (il-4), and matrix metalloproteinase 2 (mmp-2)
topic Lab/In Vitro Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4924886/
https://www.ncbi.nlm.nih.gov/pubmed/27344392
http://dx.doi.org/10.12659/MSM.896690
work_keys_str_mv AT zhangdawei fty720psuppressesosteoclastformationbyregulatingexpressionofinterleukin6il6interleukin4il4andmatrixmetalloproteinase2mmp2
AT huangyongjun fty720psuppressesosteoclastformationbyregulatingexpressionofinterleukin6il6interleukin4il4andmatrixmetalloproteinase2mmp2
AT huangzongwen fty720psuppressesosteoclastformationbyregulatingexpressionofinterleukin6il6interleukin4il4andmatrixmetalloproteinase2mmp2
AT zhangrongkai fty720psuppressesosteoclastformationbyregulatingexpressionofinterleukin6il6interleukin4il4andmatrixmetalloproteinase2mmp2
AT wanghonggang fty720psuppressesosteoclastformationbyregulatingexpressionofinterleukin6il6interleukin4il4andmatrixmetalloproteinase2mmp2
AT huangdong fty720psuppressesosteoclastformationbyregulatingexpressionofinterleukin6il6interleukin4il4andmatrixmetalloproteinase2mmp2