RESUMEN
Osteoporosis, an age associated skeletal disease, exhibits increased adipogenesis at the expense of osteogenesis from common osteoporotic bone marrow cells. In this study, black rice (Oryza sativa L.) extracts (BRE) were identified as osteogenic inducers. BRE stimulated the alkaline phosphatase (ALP) activity in both C3H10T1/2 and primary bone marrow cells. Similarly, BRE increased mRNA expression of ALP and osterix. Oral administration of BRE in OVX rats prevented decreases in bone density and strength. By contrast, BRE inhibited adipocyte differentiation of mesenchymal C3H10T1/2 cells and prevented increases in body weight and fat mass in high fat diet fed obese mice, further suggesting the dual effects of BRE on anti-adipogenesis and pro-osteogenesis. UPLC analysis identified cyanidin-3-O-glucoside and peonidin-3-O-glucoside as main anti-adipogenic effectors but not for pro-osteogenic induction. In mechanism studies, BRE selectively stimulated Wnt-driven luciferase activities. BRE treatment also induced Wnt-specific target genes such as Axin2, WISP2, and Cyclin D1. Taken together, these data suggest that BRE is a potentially useful ingredient to protect against age related osteoporosis and diet induced obesity.
Asunto(s)
Conservadores de la Densidad Ósea/uso terapéutico , Oryza/química , Osteoblastos/metabolismo , Osteogénesis , Osteoporosis Posmenopáusica/prevención & control , Extractos Vegetales/uso terapéutico , Semillas/química , Adipogénesis , Animales , Fármacos Antiobesidad/análisis , Fármacos Antiobesidad/metabolismo , Fármacos Antiobesidad/uso terapéutico , Conservadores de la Densidad Ósea/química , Conservadores de la Densidad Ósea/metabolismo , Línea Celular , Células Cultivadas , Dieta Alta en Grasa/efectos adversos , Suplementos Dietéticos , Femenino , Humanos , Masculino , Ratones Endogámicos C57BL , Obesidad/etiología , Obesidad/prevención & control , Oryza/metabolismo , Osteoblastos/citología , Osteoporosis Posmenopáusica/metabolismo , Ovariectomía , Pigmentos Biológicos/biosíntesis , Extractos Vegetales/química , Extractos Vegetales/metabolismo , Ratas Sprague-Dawley , República de Corea , Semillas/metabolismoRESUMEN
Pathological increases in adipogenic potential with decreases in osteogenic differentiation occur in osteoporotic bone marrow cells. Previous studies have shown that bioactive materials isolated from natural products can reciprocally regulate adipogenic and osteogenic fates of bone marrow cells. In this study, we showed that Eupatorium japonicum stem extracts (EJE) suppressed lipid accumulation and inhibited the expression of adipocyte markers in multipotent C3H10T1/2 and primary bone marrow cells. Conversely, EJE stimulated alkaline phosphatase activity and induced the expression of osteoblast markers in C3H10T1/2 and primary bone marrow cells. Daily oral administration of 50 mg/kg of EJE for 6 weeks to ovariectomized rats prevented body weight increase and bone mineral density decrease. Finally, activity-guided fractionation led to the identification of coumaric acid and coumaric acid methyl ester as bioactive anti-adipogenic and pro-osteogenic components in EJE. Taken together, our data indicate a promising possibility of E. japonicum as a functional food and as a therapeutic intervention for preventing osteoporosis and bone fractures.
Asunto(s)
Adipocitos/efectos de los fármacos , Enfermedades Óseas Metabólicas/prevención & control , Eupatorium/química , Células Madre Mesenquimatosas/citología , Osteoblastos/efectos de los fármacos , Osteoporosis/tratamiento farmacológico , Células 3T3 , Adipocitos/citología , Adipogénesis/efectos de los fármacos , Adiposidad/efectos de los fármacos , Fosfatasa Alcalina/metabolismo , Animales , Densidad Ósea/efectos de los fármacos , Enfermedades Óseas Metabólicas/etiología , Células de la Médula Ósea , Diferenciación Celular/efectos de los fármacos , Ácidos Cumáricos/análisis , Ácidos Cumáricos/farmacología , Femenino , Metabolismo de los Lípidos/efectos de los fármacos , Células Madre Mesenquimatosas/efectos de los fármacos , Ratones , Osteoblastos/citología , Osteoporosis/complicaciones , Extractos Vegetales/farmacología , Tallos de la Planta/química , Ratas , Ratas Sprague-DawleyRESUMEN
Sophora japonica L. fruit prevents bone loss by inhibiting osteoclast activity. We hypothesized that S japonica L. extracts could promote osteoblast differentiation. To test this hypothesis, we investigated the effect of S japonica L. on osteoblast differentiation and identified the bioactive compound(s) from S japonica L. The mature fruit of S japonica L. was partitioned with ethanol, hexane, dichloromethane (DCM), ethyl acetate, and butanol, and their effects were tested on osteoblast differentiation of C3H10T1/2 cells. DCM fractionated extracts were identified as the most osteogenic fractions. DCM fractionated extracts dose-dependently stimulated alkaline phosphatase activity and matrix mineralization. The DCM fractions also induced expression of osteoblast markers such as alkaline phosphatase, osterix, and osteocalcin in C3H10T1/2 and primary bone marrow cells. Genistein was found abundantly in the DCM fractions. Furthermore, the genistein and DCM fractions similarly modulated the expression of estrogen target genes and were both active in transfection assays that measured estrogen agonistic activity. Finally, pharmacological inhibition by treatment with an estrogen receptor antagonist or specific inhibition of gene expression by small interference RNAs targeted to estrogen receptor-ß abolished the effects of the DCM extracts, further supporting the idea that the genistein in the DCM extracts mediated the pro-osteogenic effects. Taken together, we identified genistein as the key phytoestrogen responsible for the effects of S japonica L. on osteoblast differentiation.
Asunto(s)
Expresión Génica/efectos de los fármacos , Genisteína/farmacología , Células Madre Mesenquimatosas/efectos de los fármacos , Osteoblastos/efectos de los fármacos , Osteogénesis/efectos de los fármacos , Extractos Vegetales/farmacología , Sophora/química , Fosfatasa Alcalina/metabolismo , Biomarcadores/metabolismo , Enfermedades Óseas/metabolismo , Enfermedades Óseas/prevención & control , Células de la Médula Ósea/efectos de los fármacos , Células de la Médula Ósea/metabolismo , Frutas , Humanos , Células MCF-7 , Células Madre Mesenquimatosas/metabolismo , Osteoblastos/metabolismo , Osteocalcina/metabolismo , Osteogénesis/genética , Fitoestrógenos/farmacología , Receptores de Estrógenos/metabolismoRESUMEN
Rhus verniciflua Stokes (RVS) has been used as a traditional herbal medicine for its various biological activities including anti-adipogenic effects. Activity-guided separation led to the identification of the anti-adipogenic functions of butein. Butein, a novel anti-adipogenic compound, robustly suppressed lipid accumulation and inhibited expression of adipogenic markers. Molecular studies showed that activated transforming growth factor-ß (TGF-ß) and suppressed signal transducer and activator of transcription 3 (STAT3) signaling pathways were mediated by butein. Analysis of the temporal expression profiles suggests that TGF-ß signaling precedes the STAT3 in the butein-mediated anti-adipogenic cascade. Small interfering RNA-mediated silencing of STAT3 or SMAD2/3 blunted the inhibitory effects of butein on adipogenesis indicating that an interaction between two signaling pathways is required for the action of butein. Upon butein treatments, stimulation of TGF-ß signaling was still preserved in STAT3 silenced cells, whereas regulation of STAT3 signaling by butein was significantly impaired in SMAD2/3 silenced cells, further showing that TGF-ß acts upstream of STAT3 in the butein-mediated anti-adipogenesis. Taken together, the present study shows that butein, a novel anti-adipogenic compound from RVS, inhibits adipocyte differentiation through the TGF-ß pathway followed by STAT3 and peroxisome proliferator-activated receptor γ signaling, further implicating potential roles of butein in TGF-ß- and STAT3-dysregulated diseases.