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CCN5 Reduces Ligamentum Flavum Hypertrophy by Modulating the TGF‐β Pathway
Journal of orthopaedic research, 2019-12, Vol.37 (12), p.2634-2644
Ye, Sunghyeok
Kwon, Woo‐Keun
Bae, Taegeun
Kim, Sunghyun
Lee, Jang‐Bo
Cho, Tai‐Hyoung
Park, Jung‐Yul
Kim, Kyoungmi
Hur, Junho K.
Hur, Junseok W.
2019
Volltextzugriff (PDF)
Details
Autor(en) / Beteiligte
Ye, Sunghyeok
Kwon, Woo‐Keun
Bae, Taegeun
Kim, Sunghyun
Lee, Jang‐Bo
Cho, Tai‐Hyoung
Park, Jung‐Yul
Kim, Kyoungmi
Hur, Junho K.
Hur, Junseok W.
Titel
CCN5 Reduces Ligamentum Flavum Hypertrophy by Modulating the TGF‐β Pathway
Ist Teil von
Journal of orthopaedic research, 2019-12, Vol.37 (12), p.2634-2644
Ort / Verlag
United States: John Wiley and Sons Inc
Erscheinungsjahr
2019
Quelle
Wiley-Blackwell Journals
Beschreibungen/Notizen
ABSTRACT Ligamentum flavum hypertrophy (LFH) is the most important component of lumbar spinal canal stenosis. Although the pathophysiology of LFH has been extensively studied, no method has been proposed to prevent or treat it. Since the transforming growth factor‐β (TGF‐β) pathway is known to be critical in LFH pathology, we investigated whether LFH could be prevented by blocking or modulating the TGF‐β mechanism. Human LF cells were used for the experiments. First, we created TGF‐β receptor 1 (TGFBR1) knock out (KO) cells with CRISPR (clustered regularly interspaced short palindromic repeats)/Cas9 biotechnology and treated them with TGF‐β1 to determine the effects of blocking the TGF‐β pathway. Subsequently, we studied the effect of CCN5, which has recently been proposed to modulate the TGF‐β pathway. To assess the predisposition toward fibrosis, α‐smooth muscle actin (αSMA), fibronectin, collagen‐1, collagen‐3, and CCN2 were evaluated with quantitative real‐time polymerase chain reaction, western blotting, and immunocytochemistry. The TGFBR1 KO LF cells were successfully constructed with high KO efficiency. In wild‐type (WT) cells, treatment with TGF‐β1 resulted in the overexpression of the messenger RNA (mRNA) of fibrosis‐related factors. However, in KO cells, the responses to TGF‐β1 stimulation were significantly lower. In addition, CCN5 and TGF‐β1 co‐treatment caused a notable reduction in mRNA expression levels compared with TGF‐β1 stimulation only. The αSMA protein expression increased with TGF‐β1 but decreased with CCN5 treatment. TGF‐β1 induced LF cell transdifferentiation from fibroblasts to myofibroblasts. However, this cell transition dramatically decreased in the presence of CCN5. In conclusion, CCN5 could prevent LFH by modulating the TGF‐β pathway. © 2019 The Authors. Journal of Orthopaedic Research® published by Wiley Periodicals, Inc. on behalf of Orthopaedic Research Society. J Orthop Res 37:2634–2644, 2019
Sprache
Englisch
Identifikatoren
ISSN: 0736-0266, 1554-527X
eISSN: 1554-527X
DOI: 10.1002/jor.24425
Titel-ID: cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_6899892
Format
–
Schlagworte
Actins - analysis
,
CCN Intercellular Signaling Proteins - pharmacology
,
CCN5
,
Cell Transdifferentiation - drug effects
,
Cells, Cultured
,
Clustered Regularly Interspaced Short Palindromic Repeats
,
CRISPR
,
Fibroblasts - pathology
,
Fibrosis
,
Humans
,
Hypertrophy
,
ligamentum flavum
,
Ligamentum Flavum - drug effects
,
Ligamentum Flavum - pathology
,
myofibroblast
,
Myofibroblasts - pathology
,
Receptor, Transforming Growth Factor-beta Type I - physiology
,
Repressor Proteins - pharmacology
,
Signal Transduction - physiology
,
TGF‐β1
,
Transforming Growth Factor beta - physiology
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