RESUMEN
BACKGROUND: Osteoarthritis (OA) is the most frequent articular disease and a leading cause of disability. There is a need for effective treatments able to slow the progression of disease. Some of the available treatments are dietary supplements providing natural components. Recent studies have shown that estrogen deficiency contributes to the pathophysiological events of OA progression. METHODS: We have used the anterior cruciate ligament transection model of OA in ovariectomised rats to study the effects of BIS076, a new formulation of a natural porcine cartilage extract associated with hydroxyapatite (as a source of calcium) and vitamin D3. Cartilage degradation, proteoglycan depletion and synovitis were followed by histochemistry. Effects on bone microstructure were determined by µCT. The levels of biomarkers in serum and inflammatory mediators in knee homogenates were measured by luminex or ELISA. RESULTS: Oral administration of BIS076 reduced articular cartilage damage and serum levels of cartilage degradation markers C-telopeptide of type II collagen and cartilage oligomeric matrix protein, as well as matrix metalloproteinase-3. The local inflammatory response was down-regulated by BIS076 with lower production of pro-inflammatory cytokines and prostaglandin E2 in joint tissues. In addition, BIS076 was effective on metaphyseal bone alterations as this formulation increased volumetric bone mineral density and improved bone micro-architecture. These effects were related to the modification of bone metabolism reflected by changes in bone biomarkers with reductions in the ratio receptor activator of nuclear factor κB ligand/osteoprotegerin and the levels of tartrate-resistant acid phosphatase-5b, suggesting an inhibitory activity of BIS076 on trabecular bone resorption. CONCLUSIONS: We have demonstrated the protective properties of a new formulation (BIS076) on joint lesion and bone alterations in an experimental model of OA in ovariectomised rats. This study supports the interest of BIS076 in OA treatments.
Asunto(s)
Lesiones del Ligamento Cruzado Anterior , Colágeno Tipo II/uso terapéutico , Glicosaminoglicanos/uso terapéutico , Osteoartritis de la Rodilla/tratamiento farmacológico , Osteoartritis de la Rodilla/etiología , Ovariectomía/efectos adversos , Extractos de Tejidos/uso terapéutico , Animales , Biomarcadores/sangre , Proteína de la Matriz Oligomérica del Cartílago/sangre , Colágeno Tipo II/sangre , Citocinas/sangre , Dinoprostona/sangre , Modelos Animales de Enfermedad , Durapatita/uso terapéutico , Femenino , Metaloproteinasa 3 de la Matriz/sangre , Osteoartritis de la Rodilla/sangre , Fragmentos de Péptidos/sangre , Ratas , Ratas Wistar , Porcinos , Resultado del Tratamiento , Vitamina D/uso terapéuticoRESUMEN
Age-related changes in joint tissues lead to osteoarthritis (OA). Detection of early changes in OA patients may help to initiate treatments before the establishment of irreversible joint destruction. STR/ort mice develop with age a severe degenerative joint disease that resembles human OA thus allowing the investigation of biochemical markers as well as new treatments in an accelerated time frame. We have analyzed the changes in serum levels of different mediators during the early phases of idiopathic OA in STR/ort mice. Serum levels of matrix metalloproteinase-3 (MMP-3) but not those of tumor necrosis factor-α, interleukin(IL)-1ß, IL-17 or prostaglandin E(2) correlated with histopathological changes in knees of STR/ort mice at 9 weeks. Treatment of animals with tin protoporphyrin IX (SnPP, 12 mg/kg/dayi.p.) for 4 weeks significantly reduced the progression of OA. Our data suggest that MMP-3 is a sensitive biomarker to detect early OA alterations and that SnPP could be a protective agent in OA.
Asunto(s)
Artritis Experimental/diagnóstico , Inhibidores Enzimáticos/uso terapéutico , Metaloporfirinas/uso terapéutico , Osteoartritis/diagnóstico , Protoporfirinas/uso terapéutico , Animales , Artritis Experimental/sangre , Artritis Experimental/patología , Artritis Experimental/prevención & control , Biomarcadores/sangre , Pruebas Enzimáticas Clínicas/métodos , Progresión de la Enfermedad , Evaluación Preclínica de Medicamentos/métodos , Diagnóstico Precoz , Hemo Oxigenasa (Desciclizante)/antagonistas & inhibidores , Mediadores de Inflamación/metabolismo , Masculino , Metaloproteinasa 3 de la Matriz/sangre , Ratones , Ratones Endogámicos , Osteoartritis/sangre , Osteoartritis/patología , Osteoartritis/prevención & controlRESUMEN
In a previous study, we reported a new gamma-hydroxybutenolide derivative, 4-benzo[b]thiophen-2-yl-3-bromo-5-hydroxy-5H-furan-2-one (BTH), as inhibitor of microsomal prostaglandin E synthase-1 (mPGES-1) expression in lypopolysaccharide (LPS) stimulated RAW 264.7 and TPH-1 cells, without affecting cyclooxygenase-2 (COX-2). In this study, we evaluated the in vivo effect of BTH on some acute and chronic inflammatory animal models in relation to its inhibitory profile on mPGES-1 expression. In the zymosan-induced mouse air pouch model, BTH produced a dose-dependent inhibition of prostaglandin E(2) (PGE(2)) production and mPGES-1 protein expression in pouch exudates without any effect on COX-2 protein expression. This behavior was confirmed in the chronic model of collagen-induced arthritis, where administration of BTH (5 mg/kg) clearly reduced PGE(2) and mPGES-1 expression in joint tissues, whereas COX-2 was unaffected. These effects were accompanied by the suppression of clinical and histopathological manifestations of disease such as the loss of proteoglycan, and the destruction of surface cartilage. Other enzymes participating in the metabolism of arachidonic acid, such as prostaglandin I(2) synthase, tromboxane A(2) synthase or 5-lipoxygenase were unaffected by this compound. The acetic acid-induced hyperalgesia model in LPS-sensitized mice showed a dose-dependent analgesic effect of BTH, exerting an ED(50) value of 6.2 mg/kg. Our data suggest that inhibition of mPGES-1 protein expression in acute and chronic inflammatory models by BTH, could provide a potential therapeutic target and a pharmacological tool to discern the role of the inducible enzymes COX-2 and mPGES-1 in inflammatory pathologies.
Asunto(s)
4-Butirolactona/análogos & derivados , Analgésicos/farmacología , Antiinflamatorios/farmacología , Regulación Enzimológica de la Expresión Génica/efectos de los fármacos , Oxidorreductasas Intramoleculares/metabolismo , Tiofenos/farmacología , 4-Butirolactona/farmacología , 4-Butirolactona/uso terapéutico , Acetatos/toxicidad , Analgésicos/uso terapéutico , Animales , Antiinflamatorios/uso terapéutico , Artritis Experimental/tratamiento farmacológico , Artritis Experimental/inmunología , Artritis Experimental/metabolismo , Conducta Animal/efectos de los fármacos , Plaquetas/efectos de los fármacos , Plaquetas/metabolismo , Bovinos , Enfermedad Crónica , Humanos , Hiperalgesia/inducido químicamente , Hiperalgesia/tratamiento farmacológico , Hiperalgesia/inmunología , Hiperalgesia/metabolismo , Inflamación/inducido químicamente , Inflamación/tratamiento farmacológico , Inflamación/inmunología , Leucotrieno B4/biosíntesis , Leucotrieno B4/metabolismo , Masculino , Ratones , Neutrófilos/metabolismo , Prostaglandina-E Sintasas , Tiofenos/uso terapéutico , Tromboxano B2/biosíntesis , Tromboxano B2/metabolismoRESUMEN
Avarol, a marine sesquiterpenoid hydroquinone, and 14 avarol derivatives have shown interesting anti-inflammatory properties in previous studies. In this study, avarol and derivatives were evaluated in high-throughput keratinocyte culture models using cytokeratin 10 and SKALP/Elafin expression as markers for respectively normal and psoriatic differentiation. Avarol and five of its derivatives (5, 10, 13, 14 and 15) were selected for further study. Only 10, 13, 14 and 15 were able to inhibit keratinocyte cell growth. Changes in expression levels of 22 genes were assessed by quantitative real time PCR (qPCR). From these genes, TNFalpha mRNA levels showed the strongest changes. For compound 13, 15 and dithranol (used as a model antipsoriatic drug), a dose-dependent downregulation of TNFalpha mRNA was found. The changes in TNFalpha mRNA were confirmed at the protein level for compound 13. Additionally, this compound was able to reduce also IL-8 and COX-2 mRNA levels and this effect was correlated with a reduction in COX-2 protein expression. The mechanism of action of this compound involves at least the inhibition of NF-kappaB-DNA binding activity. In conclusion, our high-throughput screening models in combination with quantitative assessment of changes in gene expression profiles identified the avarol derivative 13, a benzylamine derivative of avarol at the 4' position of benzoquinone ring, as an interesting anti-psoriatic drug candidate that inhibits keratinocyte cell growth and TNFalpha and COX-2 expression.
Asunto(s)
Antineoplásicos/farmacología , Diferenciación Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Queratinocitos/efectos de los fármacos , Psoriasis/tratamiento farmacológico , Sesquiterpenos/farmacología , Animales , Antineoplásicos/química , Células Cultivadas , Ciclooxigenasa 2/genética , Ciclooxigenasa 2/metabolismo , Evaluación Preclínica de Medicamentos , Dysidea/química , Elafina/metabolismo , Ensayo de Inmunoadsorción Enzimática , Humanos , Técnicas In Vitro , Interleucina-8/genética , Interleucina-8/metabolismo , Queratinocitos/metabolismo , Queratinocitos/patología , Queratinas/metabolismo , Proteínas de la Membrana/antagonistas & inhibidores , Proteínas de la Membrana/genética , Proteínas de la Membrana/metabolismo , Psoriasis/metabolismo , ARN Mensajero/genética , ARN Mensajero/metabolismo , Sesquiterpenos/química , Factor de Necrosis Tumoral alfa/farmacologíaRESUMEN
OBJECTIVE: Heme oxygenase 1 (HO-1) can be induced by inflammatory mediators as an adaptive response. The objective of the present study was to determine the consequences of HO-1 modulation in the murine collagen-induced arthritis (CIA) model. METHODS: DBA/1J mice were treated with an inhibitor of HO-1, tin protoporphyrin IX (SnPP), or with an inducer of HO-1, cobalt protoporphyrin IX (CoPP), from day 22 to day 29 after CIA induction. The clinical evolution of disease was monitored visually. At the end of the experiment, joints were examined for histopathologic changes. Cytokine levels in paws were measured by enzyme-linked immunosorbent assay. Levels of HO-1, cyclooxygenase 2 (COX-2), and prostaglandin E2 (PGE2) were determined. Effects of treatments on the early phase of disease and after prophylactic administration were also assessed. RESULTS: CoPP strongly induced HO-1, resulting in the inhibition of cartilage erosion accompanied by extensive fibrosis in the joint. Levels of tumor necrosis factor alpha (TNFalpha), interleukin-2 (IL-2), and IL-10 were inhibited by CoPP, whereas levels of vascular endothelial growth factor were increased. Treatment with SnPP significantly reduced the severity of CIA, with inhibition of joint inflammation and cartilage destruction. The levels of PGE2, IL-1beta, and TNFalpha were also significantly reduced by SnPP treatment, which did not modify COX-2 protein expression. SnPP was more effective than CoPP in preventing the development of CIA (prophylactic administration). CONCLUSION: HO-1 is induced during CIA. Although overexpression of this protein causes some beneficial effects, strategies aimed at HO-1 overexpression cannot slow the progression of the chronic inflammatory disease, whereas treatment with SnPP, which inhibits HO-1, exerts prophylactic and therapeutic effects.