RESUMEN
Growing data have indicated that the miR-17-92 cluster is implicated in inflammatory response and rheumatoid arthritis (RA). This study was aimed to investigate the effects of miR-92a on the proliferation and migration of rheumatoid arthritis fibroblast-like synoviocytes (RA-FLSs). Our results showed that miR-92a was significantly down-regulated in RA synovial tissue and RA-FLSs, whereas the protein level of AKT2 is increased. Restoration of miR-92a suppressed the proliferation and migration of RA-FLSs. Down-regulation of miR-92a promotes proliferation and migration of normal human FLSs. Dual luciferase reporter gene assay showed that miR-92a could specifically bind with the 30UTR of AKT2 and significantly repressed the luciferase activity. Down-regulation or up-regulation of miR-92a significantly increased or decreased the protein and phosphorylation levels of AKT2. siRNA-mediated down-regulation of AKT2 significantly prevented cell proliferation and migration of RA-FLSs, which were similar to the effects induced by overexpression of miR-92a. Moreover, AKT2 overexpression rescued miR-92a-mediated suppressive effect on proliferation and migration of RA-FLS. Thus, miR-92a could inhibit the proliferation and migration of RA-FLSs through regulation of AKT2 expression.
Asunto(s)
Artritis Reumatoide/genética , MicroARNs/genética , Proteínas Proto-Oncogénicas c-akt/genética , Sinoviocitos/metabolismo , Regiones no Traducidas 3' , Adulto , Anciano , Apoptosis , Artritis Reumatoide/metabolismo , Artritis Reumatoide/patología , Secuencia de Bases , Sitios de Unión , Estudios de Casos y Controles , Movimiento Celular , Proliferación Celular , Femenino , Regulación de la Expresión Génica , Genes Reporteros , Humanos , Luciferasas/genética , Luciferasas/metabolismo , Masculino , MicroARNs/metabolismo , Persona de Mediana Edad , Cultivo Primario de Células , Proteínas Proto-Oncogénicas c-akt/antagonistas & inhibidores , Proteínas Proto-Oncogénicas c-akt/metabolismo , ARN Interferente Pequeño/genética , ARN Interferente Pequeño/metabolismo , Transducción de Señal , Membrana Sinovial/metabolismo , Membrana Sinovial/patología , Sinoviocitos/patologíaRESUMEN
Osteoarthritis is a type of joint disease that may lead to other joint diseases. Previous research has demonstrated that tumor necrosis factor (TNF)α is associated with osteoarthritis activity and pathology. The possible mechanisms of the TNFαmediated signaling pathway have not been clearly elaborated in synovial fibroblasts. The present study aimed to investigate the potential mechanisms of TNFα in a mouse model of iodoacetateinduced osteoarthritis. Reverse transcriptionquantitative polymerase chain reaction, ELISA, western blotting and immunohistochemistry were performed to evaluate the role of TNFα in the progression of osteoarthritis. The results revealed that the serum levels of TNFα, interleukin (IL)1ß, IL4 and IL6 were significantly upregulated in a mouse model of iodoacetateinduced osteoarthritis compared with healthy mice (P<0.01). TNFα, IL1ß, IL4 and IL6 mRNA and protein levels were also significantly upregulated in synovial fibroblasts in the experimental mice (P<0.01). It was demonstrated that TNFα increased proinflammation factors matrix metalloproteinase (MMP)3, MMP9, nuclear factor (NF)κB and receptor activator of NFκB ligand (RANKL) in synovial fibroblasts. It was also observed that the tolllike receptor (TLR)3 was significantly upregulated and extracellular signalregulated kinase (ERK) and protein kinase B (AKT) were significantly downregulated in synovial fibroblasts in osteoarthritis mice (P<0.01). An in vitro assay demonstrated that TNFα inhibitor decreased mRNA and protein levels of IL1ß, IL4 and IL6 in synovial fibroblasts. The knockdown of TLR3 abolished the TNFα upregulated mRNA and protein levels of IL1ß, IL4 and IL6 in synovial fibroblasts. In addition, the knockdown of TLR3 also reversed TNFαupregulated ERK and AKT expression in synovial fibroblasts. In vivo assays demonstrated that TNFα inhibitor significantly decreased the deposition of IL1ß, IL4 and IL6 as well as bone destruction and significantly increased the body weight and osteoarthritis score for osteoarthritic mice (P<0.01). TNFα inhibitor decreased TLR3 and significantly increased the expression and phosphorylation of ERK and AKT in articular cartilage (P<0.01). In conclusion the results of the present study indicate that TNFα serves an essential role in synovial fibroblasts in osteoarthritis, suggesting that inhibition of TNFα may decrease inflammation via the TLR3mediated ERK/AKT signaling pathway in a mouse model of monosodium iodoacetateinduced osteoarthritis.
Asunto(s)
Artritis Reumatoide/genética , Artritis Reumatoide/metabolismo , Quinasas MAP Reguladas por Señal Extracelular/metabolismo , Fibroblastos/metabolismo , Proteínas Proto-Oncogénicas c-akt/metabolismo , Transducción de Señal , Receptor Toll-Like 3/metabolismo , Factor de Necrosis Tumoral alfa/metabolismo , Animales , Artritis Reumatoide/patología , Huesos/metabolismo , Citocinas/metabolismo , Modelos Animales de Enfermedad , Mediadores de Inflamación/metabolismo , Masculino , Ratones , ARN Interferente Pequeño/genética , Receptor Toll-Like 3/genéticaRESUMEN
BACKGROUND: MicroRNAs (miRs) play an important role in osteoclastogenesis. However, no study has investigated the underlying molecular mechanisms of miR-145 in this process. The purpose of the present study was to investigate the role of miR-145 and its post-transcriptional mechanism in the progression of osteoclast differentiation. METHODS: Macrophage colony stimulating factor (M-CSF) and receptor activator of nuclear factor-kB ligand (RANKL) were used to induce osteoclastogenesis originated from bone marrow-derived macrophages (BMMs). Female C57BL/6J mice were divided into sham, OVX, OVXâ¯+â¯NC-agomir and OVXâ¯+â¯miR-145-agomir groups. Tartrate-resistant acid phosphatase (TRAP) staining was performed to identify osteoclasts in-vitro and in-vivo. The mRNA and protein levels in osteoclast and tibia were assayed by qRT-PCR and western blotting, respectively. RESULTS: miR-145 expression was inhibited in RANKL-induced osteoclastogenesis, whereas overexpression of miR-145 attenuated it. We further found that Smad3 is a direct target gene of miR-145 by binding with its 3'-UTR. Overexpression of miR-145 significantly suppressed Smad3 mRNA and protein expression. In-vivo, miR-145 agomir treatment inhibited osteoclast activity in OVX mice by inhibiting Smad3 expression. CONCLUSION: We provide the evidence that over-expression of miR-145 could inhibit osteoclast differentiation, at least partially, by decreasing Smad3 expression.
Asunto(s)
Células de la Médula Ósea/metabolismo , Macrófagos/metabolismo , MicroARNs/biosíntesis , MicroARNs/genética , Osteoclastos/fisiología , Osteogénesis/genética , Ovariectomía , Ligando RANK/genética , Proteína smad3/biosíntesis , Proteína smad3/genética , Regiones no Traducidas 3'/genética , Animales , Diferenciación Celular/genética , Femenino , Factor Estimulante de Colonias de Macrófagos/biosíntesis , Ratones , Ratones Endogámicos C57BL , Ligando RANK/biosíntesis , Fosfatasa Ácida Tartratorresistente/metabolismo , Tibia/citología , Tibia/metabolismoRESUMEN
OBJECTIVE: To study the clinical efficacy of double bundle posterior cruciate ligament (PCL) reconstruction with remnant preservation. METHODS: From January 2007 to November 2011, 50 patients with PCL rupture met the inclusion criteria were divided into two groups: remnant preservation group (RP group) and remnant resection group (RR group). There were 19 males and 7 females in the RP group, ranging in age from 18 to 55 years, with a mean of (32.250 +/- 11.085) years old. The duration from injury to operation ranged from 2 to 66 months, with an average of (17.481 +/- 3.568) months. Among the RR group, 17 patients were male and 7 patients were female, ranging in age from 20 to 54 years old, with an average of (31.458 +/- 9.569) years. The duration from injury to operation ranged from 3 to 72 months, with a mean of (19.354 +/- 3.950) months. The patients in both groups suffered from instability of knee joint, got a positive result of posterior drawer test. In the RP group, the intercondylar notch remnant fiber, scar tissue and synovial were preserved in operation, only the free ligament in the intercondylar notch was resected. In the RR group, the remnant fiber, scar tissue and synovial tissue of adhesive parts were resected. In both groups, autologous semitendinosus and gracilis tendon double-bundle PCL reconstruction were carried out, the tibia was fixed with an absorbable interference screw with post-tie fixation, and the femur side was compositely fixed with absorbable interference screws and suspending fixation. Each patient received both subjective assessment (IKDC subjective evaluation, Lysholm scoring and Cincinnati rating) and objective clinical assessment (IKDC objective evaluation and Kneelax 3 tibia backward measurement) before operation and two years after operation. RESULTS: IKDC subjective evaluation: 92.167 +/- 4.177 in the RP group,which was higher than 87.542 +/- 5.687 in the RR group (P = 0.010). Lysholm scores: 90.917 +/- 4.413 in the RP group, which was higher than 87.083 +/- 5.149 in the RR group (P = 0.027). Cincinnati knee scores: 92.125 +/- 4.003 in the RP group, which was higher than 87.791 +/- 6.665 in the RR group (P = 0.027). IKDC objective evaluation:no significant statistical differences between RP group and RR group. Kneelax 3 assessment : tibia backward test with Kneelax 3 under 132 N showed no significant statistical difference between RP group and RR group, which were (3.958 +/- 0.693) mm and (4.029 +/- 0.846) mm respectively (P = 0.795). CONCLUSION: The study shows a significant advantage of remnant fiber preservation than remnant fiber resection in double-bundle PCL construction in terms of subjective knee function recovery after operation. There is no significant difference in postoperative knee stability.
Asunto(s)
Articulación de la Rodilla/cirugía , Procedimientos de Cirugía Plástica/métodos , Ligamento Cruzado Posterior/cirugía , Adolescente , Adulto , Artroscopía , Estudios de Casos y Controles , Femenino , Humanos , Masculino , Persona de Mediana Edad , Adulto JovenRESUMEN
OBJECTIVE: To evaluate the effectiveness of open reduction and internal fixation and repair of palmar ligment in treating trans-scaphoid perilunate dislocation. METHODS: From June 1995 to June 2001,14 patients with trans-scaphoid perilunate dislocation were treated with open reduction and internal fixation and repair of palmar ligment. Among them,there were 13 males and 1 female,the ranging in age from 21 to 38 years,averaged 25.4 years. All patients were posterior dislocation and all operations were performed within 2 weeks after injury. RESULTS: All patients were followed up from 24 to 60 months with an average of 28.3 months. Thirteen scaphoid fractures were primary healed and functions of wrist joint were good. Bone disunion was found in 1 case and part functions of wrist joint were limited. No found necrosis of lunate and scaphoid. According to clinical scoring system of Cooney, 9 case got excellent results, 3 good, 1 fair and 1 poor. CONCLUSION: Open reduction and internal fixation and repair of palmar ligament is effective in treating trans-scaphoid perilunate dislocation,which can early provide steady fixation for scaphoid,and profit to recover blood supply of lunatum and subterminal scaphoid.