Dysregulated circadian rhythm pathway in human osteoarthritis: NR1D1 and BMAL1 suppression alters TGF-ß signaling in chondrocytes.
Osteoarthritis Cartilage
; 25(6): 943-951, 2017 06.
Article
en En
| MEDLINE
| ID: mdl-27884645
ABSTRACT
OBJECTIVES:
Circadian rhythm (CR) was identified by RNA sequencing as the most dysregulated pathway in human osteoarthritis (OA) in articular cartilage. This study examined circadian rhythmicity in cultured chondrocytes and the role of the CR genes NR1D1 and BMAL1 in regulating chondrocyte functions.METHODS:
RNA was extracted from normal and OA-affected human knee cartilage (n = 14 each). Expression levels of NR1D1 and BMAL1 mRNA and protein were assessed by quantitative PCR and immunohistochemistry. Human chondrocytes were synchronized and harvested at regular intervals to examine circadian rhythmicity in RNA and protein expression. Chondrocytes were treated with small interfering RNA (siRNA) for NR1D1 or BMAL1, followed by RNA sequencing and analysis of the effects on the transforming growth factor beta (TGF-ß) pathway.RESULTS:
NR1D1 and BMAL1 mRNA and protein levels were significantly reduced in OA compared to normal cartilage. In cultured human chondrocytes, a clear circadian rhythmicity was observed for NR1D1 and BMAL1. Increased BMAL1 expression was observed after knocking down NR1D1, and decreased NR1D1 levels were observed after knocking down BMAL1. Sequencing of RNA from chondrocytes treated with NR1D1 or BMAL1 siRNA identified 330 and 68 significantly different genes, respectively, and this predominantly affected the TGF-ß signaling pathway.CONCLUSIONS:
The CR pathway is dysregulated in OA cartilage. Interference with circadian rhythmicity in cultured chondrocytes affects TGF-ß signaling, which is a central pathway in cartilage homeostasis.Palabras clave
Texto completo:
1
Banco de datos:
MEDLINE
Asunto principal:
ARN Mensajero
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Cartílago Articular
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Factor de Crecimiento Transformador beta
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Ritmo Circadiano
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Condrocitos
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Osteoartritis de la Rodilla
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Factores de Transcripción ARNTL
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Miembro 1 del Grupo D de la Subfamilia 1 de Receptores Nucleares
Tipo de estudio:
Prognostic_studies
Límite:
Adolescent
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Adult
/
Female
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Humans
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Male
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Middle aged
Idioma:
En
Revista:
Osteoarthritis Cartilage
Asunto de la revista:
ORTOPEDIA
/
REUMATOLOGIA
Año:
2017
Tipo del documento:
Article
País de afiliación:
Japón