RESUMEN
Glucocorticoid provokes bone mass loss and fatty marrow, accelerating osteoporosis development. Bromodomain protein BRD4, an acetyl-histone-binding chromatin reader, regulates stem cell and tissue homeostasis. We uncovered that glucocorticoid inhibited acetyl Lys-9 at the histone 3 (H3K9ac)-binding Runx2 promoter and decreased osteogenic differentiation, whereas bromodomain protein 4 (BRD4) and adipocyte formation were upregulated in bone-marrow mesenchymal progenitor cells. BRD4 knockdown improved H3K9ac occupation at the Runx2 promoter and osteogenesis, but attenuated glucocorticoid-mediated adipocyte formation together with the unaffected H3K9ac-binding PPARγ2 promoter. BRD4 regulated epigenome related to fatty acid metabolism and the forkhead box P1 (Foxp1) pathway, which occupied the PPARγ2 promoter to modulate glucocorticoid-induced adipocytic activity. In vivo, BRD4 inhibitor JQ-1 treatment mitigated methylprednisolone-induced suppression of bone mass, trabecular microstructure, mineral acquisition, and osteogenic differentiation. Foxp1 signaling, marrow fat, and adipocyte formation in glucocorticoid-treated skeleton were reversed upon JQ-1 treatment. Taken together, glucocorticoid-induced H3K9 hypoacetylation augmented BRD4 action to Foxp1, which steered mesenchymal progenitor cells toward adipocytes at the cost of osteogenic differentiation in osteoporotic skeletons. BRD4 inhibition slowed bone mass loss and marrow adiposity. Collective investigations convey a new epigenetic insight into acetyl histone reader BRD4 control of osteogenesis and adipogenesis in skeleton, and highlight the remedial effects of the BRD4 inhibitor on glucocorticoid-induced osteoporosis.
Asunto(s)
Adipogénesis/fisiología , Médula Ósea/metabolismo , Proteínas de Ciclo Celular/metabolismo , Glucocorticoides/metabolismo , Factores de Transcripción/metabolismo , Diferenciación Celular/efectos de los fármacos , Glucocorticoides/farmacología , Humanos , Células Madre Mesenquimatosas/metabolismo , Osteoblastos/efectos de los fármacos , Osteogénesis/fisiologíaRESUMEN
Rotator cuff lesion with shoulder stiffness is a major cause of shoulder pain and motionlessness. Subacromial bursa fibrosis is a prominent pathological feature of the shoulder disorder. MicroRNA-29a (miR-29a) regulates fibrosis in various tissues; however, the miR-29a action to subacromial bursa fibrosis remains elusive. Here, we reveal that subacromial synovium in patients with rotator cuff tear with shoulder stiffness showed severe fibrosis, hypertrophy, and hyperangiogenesis histopathology along with significant increases in fibrotic matrices collagen (COL) 1A1, 3A1, and 4A1 and inflammatory cytokines, whereas miR-29a expression was downregulated. Supraspinatus and infraspinatus tenotomy-injured shoulders in transgenic mice overexpressing miR-29a showed mild swelling, vascularization, fibrosis, and regular gait profiles as compared to severe rotator cuff damage in wild-type mice. Treatment with miR-29a precursor compromised COL3A1 production and hypervascularization in injured shoulders. In vitro, gain of miR-29a function attenuated COL3A1 expression through binding to the 3'-untranslated region (3'-UTR) of COL3A1 in inflamed tenocytes, whereas silencing miR-29a increased the matrix expression. Taken together, miR-29a loss is correlated with subacromial bursa inflammation and fibrosis in rotator cuff tear with shoulder stiffness. miR-29a repressed subacromial bursa fibrosis through directly targeting COL3A1 mRNA, improving rotator cuff integrity and shoulder function. Collective analysis offers a new insight into the molecular mechanism underlying rotator cuff tear with shoulder stiffness. This study also highlights the remedial potential of miR-29a precursor for alleviating the shoulder disorder.
Asunto(s)
Bolsa Sinovial/metabolismo , MicroARNs/metabolismo , Lesiones del Manguito de los Rotadores/metabolismo , Anciano , Anciano de 80 o más Años , Animales , Bolsa Sinovial/patología , Bursitis/metabolismo , Bursitis/patología , Femenino , Humanos , Artropatías/metabolismo , Artropatías/patología , Ratones , MicroARNs/genética , Persona de Mediana Edad , Lesiones del Manguito de los Rotadores/patologíaRESUMEN
Glucocorticoid excess escalates osteoclastic resorption, accelerating bone mass loss and microarchitecture damage, which ramps up osteoporosis development. MicroRNA-29a (miR-29a) regulates osteoblast and chondrocyte function; however, the action of miR-29a to osteoclastic activity in the glucocorticoid-induced osteoporotic bone remains elusive. In this study, we showed that transgenic mice overexpressing an miR-29a precursor driven by phosphoglycerate kinase exhibited a minor response to glucocorticoid-mediated bone mineral density loss, cortical bone porosity and overproduction of serum resorption markers C-teleopeptide of type I collagen and tartrate-resistant acid phosphatase 5b levels. miR-29a overexpression compromised trabecular bone erosion and excessive osteoclast number histopathology in glucocorticoid-treated skeletal tissue. Ex vivo, the glucocorticoid-provoked osteoblast formation and osteoclastogenic markers (NFATc1, MMP9, V-ATPase, carbonic anhydrase II and cathepsin K) along with F-actin ring development and pit formation of primary bone-marrow macrophages were downregulated in miR-29a transgenic mice. Mechanistically, tumor necrosis factor superfamily member 13b (TNFSF13b) participated in the glucocorticoid-induced osteoclast formation. miR-29a decreased the suppressor of cytokine signaling 2 (SOCS2) enrichment in the TNFSF13b promoter and downregulated the cytokine production. In vitro, forced miR-29a expression and SOCS2 knockdown attenuated the glucocorticoid-induced TNFSF13b expression in osteoblasts. miR-29a wards off glucocorticoid-mediated excessive bone resorption by repressing the TNFSF13b modulation of osteoclastic activity. This study sheds new light onto the immune-regulatory actions of miR-29a protection against glucocorticoid-mediated osteoporosis.
Asunto(s)
Factor Activador de Células B/metabolismo , Resorción Ósea/genética , MicroARNs/fisiología , Osteogénesis/genética , Animales , Factor Activador de Células B/genética , Hueso Esponjoso/patología , Diferenciación Celular , Inmunohistoquímica , Ratones Transgénicos , MicroARNs/genética , MicroARNs/metabolismo , Osteoclastos/patología , Transducción de SeñalRESUMEN
Osteoporosis deteriorates bone mass and biomechanical strength, becoming a life-threatening cause to the elderly. MicroRNA is known to regulate tissue remodeling; however, its role in the development of osteoporosis remains elusive. In this study, we uncovered that silencing miR-29a expression decreased mineralized matrix production in osteogenic cells, whereas osteoclast differentiation and pit formation were upregulated in bone marrow macrophages as co-incubated with the osteogenic cells in transwell plates. In vivo, decreased miR-29a expression occurred in ovariectomy-mediated osteoporotic skeletons. Mice overexpressing miR-29a in osteoblasts driven by osteocalcin promoter (miR-29aTg/OCN) displayed higher bone mineral density, trabecular volume and mineral acquisition than wild-type mice. The estrogen deficiency-induced loss of bone mass, trabecular morphometry, mechanical properties, mineral accretion and osteogenesis of bone marrow mesenchymal cells were compromised in miR-29aTg/OCN mice. miR-29a overexpression also attenuated the estrogen loss-mediated excessive osteoclast surface histopathology, osteoclast formation of bone marrow macrophages, receptor activator nuclear factor-κ ligand (RANKL) and C-X-C motif chemokine ligand 12 (CXCL12) expression. Treatment with miR-29a precursor improved the ovariectomy-mediated skeletal deterioration and biomechanical property loss. Mechanistically, miR-29a inhibited RANKL secretion in osteoblasts through binding to 3'-UTR of RANKL. It also suppressed the histone acetyltransferase PCAF-mediated acetylation of lysine 27 in histone 3 (H3K27ac) and decreased the H3K27ac enrichment in CXCL12 promoters. Taken together, miR-29a signaling in osteogenic cells protects bone tissue from osteoporosis through repressing osteoclast regulators RANKL and CXCL12 to reduce osteoclastogenic differentiation. Arrays of analyses shed new light on the miR-29a regulation of crosstalk between osteogenic and osteoclastogenic cells. We also highlight that increasing miR-29a function in osteoblasts is beneficial for bone anabolism to fend off estrogen deficiency-induced excessive osteoclastic resorption and osteoporosis.
Asunto(s)
Quimiocina CXCL12/genética , MicroARNs/metabolismo , Osteoclastos/metabolismo , Osteoporosis/genética , Ligando RANK/genética , Factores de Transcripción p300-CBP/metabolismo , Animales , Fenómenos Biomecánicos , Huesos/citología , Huesos/metabolismo , Comunicación Celular/fisiología , Diferenciación Celular/fisiología , Femenino , Humanos , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Transgénicos , MicroARNs/genética , Osteoclastos/citología , Ovariectomía , Ligando RANK/metabolismo , Factores de Transcripción p300-CBP/genéticaRESUMEN
Hard ticks infesting canine hosts were determined for the first time in northern Taiwan. Between May 2010 and April 2011, a total of 9467 ticks were collected from 2025 dogs. They were identified based on pictorial keys of their morphological characteristics. These ticks belong to three genus and six species: Rhipicephalus sanguineus, R. haemaphysaloides, Haemaphysalis hystricis, H. lagrangei, H. formosensis and Ixodes ovatus. Rhipicephalus sanguineus was the most dominant species (92.5%), followed by H. hystricis (4.6%), R. haemaphysaloides (2.3%), I. ovatus (0.54%), H. lagrangei (0.04%) and H. formosensis (0.01%). The overall density was 4.7 ticks per dog (ranging from 1.8 to 20.8) and the highest seasonal prevalence was observed in September with an average density of 8.2 ticks per dog. Our results not only provide the first survey of hard ticks infesting dogs in northern Taiwan, but also highlight the possible impact of these tick species on human health.
Asunto(s)
Enfermedades de los Perros/epidemiología , Ixodidae/fisiología , Infestaciones por Garrapatas/veterinaria , Animales , Enfermedades de los Perros/parasitología , Perros , Femenino , Ixodidae/crecimiento & desarrollo , Larva/crecimiento & desarrollo , Larva/fisiología , Masculino , Ninfa/crecimiento & desarrollo , Ninfa/fisiología , Taiwán/epidemiología , Infestaciones por Garrapatas/epidemiología , Infestaciones por Garrapatas/parasitologíaRESUMEN
A total of 578 Rhipicephalus sanguineus ticks collected from dogs in Taiwan were examined for Babesia by species-specific polymerase chain reaction assay based on the 18S small subunit ribosomal RNA (ssrRNA) gene. Babesia DNA was detected in 1.04 % (6/578) of Rh. sanguineus ticks. Phylogenetic analysis revealed that these Babesia spp. were genetically linked to the same clade within the genospecies of Babesia vogeli and could be discriminated from other genospecies of Babesia. Intra-species analysis based on the genetic distance values indicated a lower level (0.079) compared with other genospecies of Babesia (GD > 0.094) and out-group protozoa (GD > 0.236). This study provides the first molecular evidence of B. vogeli detected and identified in various stages of Rh. sanguineus ticks removed from dogs in Taiwan. Detection of Rh. sanguineus in flat male ticks may imply the possible mechanism of transstadial transmission in Rh. sanguineus ticks. The vector competence and the diversity of Babesia species harbored by Rh. sanguineus ticks need to be further investigated.
Asunto(s)
Babesia/aislamiento & purificación , Rhipicephalus sanguineus/parasitología , Animales , Babesia/clasificación , Babesia/genética , Babesiosis/diagnóstico , Enfermedades de los Perros/parasitología , Perros , Femenino , Masculino , Filogenia , Reacción en Cadena de la Polimerasa/veterinaria , TaiwánRESUMEN
A Taiwan stink snake (Elaphe carinata) was captured in shrubbery at a fruit farm in southern Taiwan, and 3 female ticks (2 flat, one fully engorged) observed on this snake were collected and identified as Amblyomma helvolum, the first record of this host association in mainland Taiwan. The key morphological characters of female A. helvolum were photographed using a stereomicroscope. This first report of A. helvolum feeding on a reptilian host in Taiwan includes the first color illustrations of adult A. helvolum ever published. The increased international trade in wild reptiles may contribute to accidental transportation of ticks into Taiwan. The host range and vector capacity of A. helvolum for various tick-borne pathogens need to be further clarified.