RESUMEN
PURPOSE: Hashimoto's thyroiditis (HT) is a common autoimmune thyroid disorder that can disrupt thyroid function and homeostasis. As HT results from a dysregulated immune system, we hypothesized that these patients might be more susceptible to transplant failure; however, literature on this association is limited. The purpose of this study is to examine the association of HT with the risk of renal transplant failure. METHODS: We utilized the United States Renal Database System dataset collected from 2005 to 2014 and compared the time from first renal transplant to transplant failure in end-stage renal disease (ESRD) patients with a HT diagnosis to ESRD patients without a HT diagnosis that underwent renal transplant. RESULTS: A total of 144 ESRD patients had International Classification of Disease-9 claim codes for HT prior to renal transplant, amongst a total cohort of 90,301 renal transplant patients aged 18-100 and meeting criteria. Patients with HT were significantly more likely to be female, white, and to have a diagnosis of cytomegalovirus compared to patients without. ESRD patients with a HT diagnosis that underwent renal transplant had a significantly increased risk of renal transplant failure compared to those ESRD renal transplant patients without an HT diagnosis. There was a significantly increased adjusted hazard ratio for graft failure in patients with a HT diagnosis compared to those without. CONCLUSION: Thyroid health and HT may play a significant role in the development of the increased risk of renal transplant failure observed in this study. Additional studies are needed to investigate the underlying mechanisms for this association.
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Enfermedad de Hashimoto , Enfermedades Renales , Fallo Renal Crónico , Trasplante de Riñón , Humanos , Femenino , Masculino , Trasplante de Riñón/efectos adversos , Enfermedad de Hashimoto/complicaciones , Fallo Renal Crónico/etiología , Fallo Renal Crónico/cirugíaRESUMEN
The plasma n-3 fatty acid level was 26.2% lower in patients with osteoporotic hip fracture than in those with osteoarthritis. In all patients, n-3 fatty acid was positively associated with bone mineral density and inversely associated with tartrate-resistant acid phosphatase-5b level in bone marrow aspirates, reflecting the bone microenvironment. INTRODUCTION: Despite the potential beneficial role of n-3 fatty acid (FA) on bone metabolism, the specific mechanisms underlying these effects in humans remain unclear. Here, we assessed whether the plasma n-3 level, as an objective indicator of its status, is associated with osteoporosis-related phenotypes and bone-related markers in human bone marrow (BM) samples. METHODS: This was a case-control and cross-sectional study conducted in a clinical unit. n-3 FA in the blood and bone biochemical markers in the BM aspirates were measured by gas chromatography/mass spectrometry and immunoassay, respectively. BM fluids were collected from 72 patients who underwent hip surgery because of either osteoporotic hip fracture (HF; n = 28) or osteoarthritis (n = 44). RESULTS: After adjusting for confounders, patients with HF had 26.2% lower plasma n-3 levels than those with osteoarthritis (P = 0.006), and each standard deviation increment in plasma n-3 was associated with a multivariate-adjusted odds ratio of 0.40 for osteoporotic HF (P = 0.010). In multivariate analyses including all patients, a higher plasma n-3 level was associated with higher bone mass at the lumbar spine (ß = 0.615, P = 0.002) and total femur (ß = 0.244, P = 0.045). Interestingly, the plasma n-3 level was inversely associated with the tartrate-resistant acid phosphatase-5b level (ß = - 0.633, P = 0.023), but not with the bone-specific alkaline phosphatase level, in BM aspirates. CONCLUSIONS: These findings provide clinical evidence that n-3 FA is a potential inhibitor of osteoclastogenesis that favors human bone health.
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Densidad Ósea/fisiología , Ácidos Grasos Omega-3/sangre , Fracturas de Cadera/fisiopatología , Fracturas Osteoporóticas/fisiopatología , Fosfatasa Ácida Tartratorresistente/metabolismo , Anciano , Anciano de 80 o más Años , Médula Ósea/metabolismo , Resorción Ósea/fisiopatología , Estudios de Casos y Controles , Estudios Transversales , Ácidos Grasos Omega-3/fisiología , Ácidos Grasos Omega-6/sangre , Femenino , Fémur/fisiopatología , Fracturas de Cadera/sangre , Humanos , Vértebras Lumbares/fisiopatología , Masculino , Fracturas Osteoporóticas/sangreRESUMEN
Data gathered from a nationally representative cohort demonstrate that higher dietary protein intake was positively associated with the composite indices of femoral neck strength in both men and women, suggesting that higher protein intake may contribute to lower risk of hip fracture through the improvement of bone strength. INTRODUCTION: Despite the general belief that higher protein intake may be helpful for bone homeostasis, its impact on human bone health is still debated. Furthermore, the association of dietary protein intake with femoral neck (FN) strength, which can predict fracture risk independently of bone mineral density (BMD), has not been thoroughly studied. METHODS: This is a population-based, cross-sectional study from Korea National Health and Nutrition Examination Surveys, including 592 men aged 50 years or older and 590 postmenopausal women. The composite indices of FN strength, such as the compression strength index (CSI), bending strength index (BSI), and impact strength index (ISI), were generated by combining BMD, body weight, and height with the femoral axis length and width, which were measured by dual-energy X-ray absorptiometry. RESULTS: After adjustment for confounders, total protein intake (g/kg/day) positively correlated with all three FN composite indices in both genders (P = 0.006 to 0.035), except for BSI showing marginal significance in postmenopausal women (P = 0.093). Consistently, compared with subjects in lowest total protein intake quartile, those in the highest quartile showed markedly higher CSI, BSI, and ISI values (P = 0.043 to < 0.001), with a dose-response manner across increasing total protein intake quartile categories in both men and women (P for trend = 0.028 to < 0.001). CONCLUSIONS: These findings provide the clinical evidence that higher dietary protein intake can play a beneficial role on bone health through the increase of FN strength relative to load in adults.
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Proteínas en la Dieta/farmacología , Cuello Femoral/efectos de los fármacos , Absorciometría de Fotón/métodos , Anciano , Anciano de 80 o más Años , Antropometría/métodos , Estatura/fisiología , Peso Corporal/fisiología , Densidad Ósea/efectos de los fármacos , Estudios Transversales , Dieta/estadística & datos numéricos , Proteínas en la Dieta/administración & dosificación , Femenino , Cuello Femoral/fisiología , Humanos , Masculino , Persona de Mediana Edad , Encuestas Nutricionales , Caracteres SexualesRESUMEN
There was no association of plasma DPP-4 activity levels with bone mineral density (BMD), body composition, or incident hip fractures in a cohort of elderly community-dwelling adults. INTRODUCTION: Dipeptidyl peptidase IV (DPP-4) inactivates several key hormones including those that stimulate postprandial insulin secretion, and DPP-4 inhibitors (gliptins) are approved to treat diabetes. While DPP-4 is known to modulate osteogenesis, the relationship between DPP-4 activity and skeletal health is uncertain. The purpose of the present study was to examine possible associations between DPP-4 activity in elderly subjects enrolled in the Cardiovascular Health Study (CHS) and BMD, body composition measurements, and incident hip fractures. METHODS: All 1536 male and female CHS participants who had evaluable DXA scans and plasma for DPP-4 activity were included in the analyses. The association between (1) BMD of the total hip, femoral neck, lumbar spine, and total body; (2) body composition measurements (% lean, % fat, and total body mass); and (3) incident hip fractures and plasma levels of DPP-4 activity were determined. RESULTS: Mean plasma levels of DPP-4 activity were significantly higher in blacks (227 ± 78) compared with whites (216 ± 89) (p = 0.04). However, there was no significant association of DPP-4 activity with age or gender (p ≥ 0.14 for both). In multivariable adjusted models, there was no association of plasma DPP-4 activity with BMD overall (p ≥ 0.55 for all) or in gender stratified analyses (p ≥ 0.23). There was also no association of DPP-4 levels and incident hip fractures overall (p ≥ 0.24) or in gender stratified analyses (p ≥ 0.39). CONCLUSION: Plasma DPP-4 activity, within the endogenous physiological range, was significantly associated with race, but not with BMD, body composition, or incident hip fractures in elderly community-dwelling subjects.
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Composición Corporal/fisiología , Densidad Ósea/fisiología , Dipeptidil Peptidasa 4/sangre , Fracturas de Cadera/sangre , Anciano , Anciano de 80 o más Años , Población Negra/estadística & datos numéricos , Diabetes Mellitus/sangre , Diabetes Mellitus/etnología , Diabetes Mellitus/fisiopatología , Dipeptidil Peptidasa 4/fisiología , Femenino , Fracturas de Cadera/etnología , Fracturas de Cadera/fisiopatología , Humanos , Incidencia , Estudios Longitudinales , Masculino , Factores Sexuales , Estados Unidos/epidemiología , Población Blanca/estadística & datos numéricosRESUMEN
CONTEXT: IGF-1 promotes bone growth directly and indirectly through its effects on skeletal muscle. Insulin and IGF-1 share a common cellular signaling process; thus, insulin resistance may influence the IGF-1-muscle-bone relationship. OBJECTIVE: We sought to determine the effect of insulin resistance on the muscle-dependent relationship between IGF-1 and bone mass in premenarcheal girls. DESIGN, SETTING, AND PARTICIPANTS: This was a cross-sectional study conducted at a university research center involving 147 girls ages 9 to 11 years. MAIN OUTCOME MEASURES: Glucose, insulin, and IGF-1 were measured from fasting blood samples. Homeostasis model assessment of insulin resistance (HOMA-IR) was calculated from glucose and insulin. Fat-free soft tissue (FFST) mass and bone mineral content (BMC) were measured by dual-energy x-ray absorptiometry. Our primary outcome was BMC/height. RESULTS: In our path model, IGF-1 predicted FFST mass (b = 0.018; P = .001), which in turn predicted BMC/height (b = 0.960; P < .001). IGF-1 predicted BMC/height (b = 0.001; P = .002), but not after accounting for the mediator of this relationship, FFST mass. The HOMA-IR by IGF-1 interaction negatively predicted FFST mass (b = -0.044; P = .034). HOMA-IR had a significant and negative effect on the muscle-dependent relationship between IGF-1 and BMC/height (b = -0.151; P = .047). CONCLUSIONS: Lean body mass is an important intermediary factor in the IGF-1-bone relationship. For this reason, bone development may be compromised indirectly via suboptimal IGF-1-dependent muscle development in insulin-resistant children.
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Densidad Ósea/fisiología , Resistencia a la Insulina/fisiología , Factor I del Crecimiento Similar a la Insulina/metabolismo , Músculo Esquelético/fisiología , Absorciometría de Fotón , Glucemia/metabolismo , Composición Corporal/fisiología , Estatura , Niño , Estudios Transversales , Método Doble Ciego , Femenino , Humanos , Insulina/sangre , Factor I del Crecimiento Similar a la Insulina/genética , Menarquia , Músculo Esquelético/anatomía & histologíaRESUMEN
Muscle- and liver-derived IGF-1 play important roles in muscle anabolism throughout growth and aging. Yet, prolonged food restriction is thought to increase longevity in part by lowering levels of IGF-1, which in turn reduces the risk for developing various cancers. The dietary factors that modulate IGF-1 levels are, however, poorly understood. We tested the hypothesis that the adipokine leptin, which is elevated with food intake and suppressed during fasting, is a key mediator of IGF-1 levels with aging and food restriction. First, leptin levels in peripheral tissues were measured in young mice fed ad libitum, aged mice fed ad libitum, and aged calorie-restricted (CR) mice. A group of aged CR mice were also treated with recombinant leptin for 10 days. Later, aged mice fed ad libitum were treated with saline (VEH) or with a novel leptin receptor antagonist peptide (Allo-aca) and tissue-specific levels of IGF-1 were determined. On one hand, recombinant leptin induced a three-fold increase in liver-derived IGF-1 and a two-fold increase in muscle-derived IGF-1 in aged, CR mice. Leptin also significantly increased serum growth hormone levels in the aged, CR mice. On the other, the leptin receptor antagonist Allo-aca did not alter body weight or muscle mass in treated mice compared to VEH mice. Allo-aca did, however, produce a significant (20%) decline in liver-derived IGF-1 as well as an even more pronounced (>50%) decrease in muscle-derived IGF-1 compared to VEH-treated mice. The reduced IGF-1 levels in Allo-aca treated mice were not accompanied by any significant change in growth hormone levels compared to VEH mice. These findings suggest that leptin receptor antagonists may represent novel therapeutic agents for attenuating IGF-1 signaling associated with aging, and could potentially mimic some of the positive effects of calorie restriction on longevity.
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Envejecimiento/metabolismo , Factor I del Crecimiento Similar a la Insulina/metabolismo , Leptina/fisiología , Hígado/metabolismo , Músculo Esquelético/metabolismo , Animales , Peso Corporal/efectos de los fármacos , Peso Corporal/fisiología , Restricción Calórica , Ingestión de Alimentos , Hormona del Crecimiento/sangre , Factor I del Crecimiento Similar a la Insulina/efectos de los fármacos , Leptina/farmacología , Longevidad/fisiología , Ratones , Péptidos/farmacología , Receptores de Leptina/antagonistas & inhibidores , Proteínas Recombinantes/farmacologíaRESUMEN
A commercial resin-based pine oil (PO) derived from Pinus palustris and Pinus elliottii was the major focus of this investigation. Extracts of pine resins, needles, and bark are folk medicines commonly used to treat skin ailments, including burns. The American Burn Association estimates that 500,000 people with burn injuries receive medical treatment each year; one-half of US burn victims are children, most with scald burns. This systematic study was initiated as follow-up to personal anecdotal evidence acquired over more than 10 years by MH Bhattacharyya regarding PO's efficacy for treating burns. The results demonstrate that PO counteracted dermal inflammation in both a mouse ear model of contact irritant-induced dermal inflammation and a second degree scald burn to the mouse paw. Furthermore, PO significantly counteracted the tactile allodynia and soft tissue injury caused by the scald burn. In mouse dorsal root ganglion neuronal cultures, PO added to the medium blocked adenosine triphosphate-activated, but not capsaicin-activated, pain pathways, demonstrating specificity. These results together support the hypothesis that a pine-oil-based treatment can be developed to provide effective in-home care for second degree burns.
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Quemaduras/tratamiento farmacológico , Ganglios Espinales/efectos de los fármacos , Pinus/química , Aceites de Plantas/farmacología , Adenosina Trifosfato , Animales , Capsaicina , Células Cultivadas , Dermatitis/tratamiento farmacológico , Modelos Animales de Enfermedad , Hiperalgesia/tratamiento farmacológico , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Endogámicos ICR , Neuronas/efectos de los fármacos , Neuronas/metabolismo , Dolor/tratamiento farmacológico , Resinas de Plantas/farmacología , Piel/patologíaRESUMEN
CONTEXT: The extent to which 25-hydroxyvitamin D [25(OH)D] and IGF-I influence bone mineral content (BMC) accrual from early to mid-puberty is unclear. OBJECTIVE, SETTING, AND PARTICIPANTS: This study sought to determine relationships among 25(OH)D, IGF-I, and BMC in community-dwelling prepubertal females (n = 76; aged 4-8 yr at baseline) over a period of up to 9 yr. DESIGN: The hypothesis that changes in IGF-I vs. 25(OH)D are more strongly associated with BMC accrual was formulated after data collection. 25(OH)D and IGF-I were log-transformed and further adjusted using two-way ANOVA for differences in season and race. Linear mixed modeling (including a random subject-specific intercept and a random subject-specific slope on age) was employed to analyze the proportion of variance the transformed 25(OH)D and IGF-I variables explained for the bone outcomes. RESULTS: IGF-I was more strongly associated with BMC accrual than 25(OH)D at the total body (R(2) = 0.874 vs. 0.809), proximal femur (R(2) = 0.847 vs. 0.771), radius (R(2) = 0.812 vs. 0.759), and lumbar spine (R(2) = 0.759 vs. 0.698). The rate of BMC accrual was positively associated with changes in IGF-I but negatively associated with 25(OH)D. When IGF-I and 25(OH)D were included in the same regression equation, 25(OH)D did not have a significant predictive effect on BMC accrual above and beyond that of IGF-I. CONCLUSIONS: These prospective data in early adolescent females indicate that both 25(OH)D and IGF-I have a significant impact on bone mineral accrual; however, the positive association of IGF-I and BMC accrual is greater than the negative association of 25(OH)D and BMC accrual.
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Densidad Ósea/fisiología , Calcificación Fisiológica/fisiología , Factor I del Crecimiento Similar a la Insulina/metabolismo , Vitamina D/análogos & derivados , Análisis de Varianza , Composición Corporal , Niño , Preescolar , Dieta , Femenino , Humanos , Modelos Lineales , Estudios Prospectivos , Vitamina D/sangreRESUMEN
The regulation of bone metabolism mediated by leptin is a complex process that is not clearly understood. Recent studies suggest that CART (cocaine-amphetamine related transcript) is a significant neuronal co-factor when combined with leptin. CART deficiency is thought to result in low trabecular bone mass, but since leptin exerts contrasting effects on trabecular and cortical bone it is possible that cortical bone may not respond to the absence of CART signaling in the same manner as trabecular bone. We tested the hypothesis that CART deficiency decreases cortical bone mass, density, and strength by examining femora of adult wild-type mice (CART(+/+)) and CART-deficient mice (CART(-/-)). DEXA densitometry (PIXImus system) was used to measure whole-bone mineral content (BMC) and mineral density (BMD) from right femora, and pQCT used to calculate densitometric and geometric parameters from the femur midshaft. Femora were also tested in three-point bending, and sections of the tibia analyzed histologically to determine bone marrow adipocyte density (N.At./M.Ar) and endocortical osteoclast number (N.Oc/B.Pm). The control mice weighed less than the mice lacking CART (P<0.001), but mechanical testing data showed no differences (p>0.05) in ultimate force, energy to fracture, stiffness, or intrinsic properties such as ultimate stress, ultimate strain, or modulus. CART-deficient mice did not differ from normal controls in whole-femur BMC (p=0.09), BMD (p=0.19), midshaft cortical bone thickness (p=0.67), midshaft cortical bone area (p=0.59) or N.Oc/B.Pm (p=0.94), although CART deficiency was associated with a three-fold increase in bone marrow adipocyte density (p<0.001). Our data suggest that while the central, neuroendocrine regulation of bone mass via CART signaling may have effects on trabecular mass, absence of CART expression does not significantly alter cortical bone geometry, density, or strength.
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Peso Corporal , Huesos/fisiopatología , Proteínas del Tejido Nervioso/deficiencia , Resistencia a la Tracción , Absorciometría de Fotón , Adipocitos/patología , Animales , Densidad Ósea , Médula Ósea/patología , Recuento de Células , Fémur/patología , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones NoqueadosAsunto(s)
Remodelación Ósea/fisiología , Hormonas Gastrointestinales/fisiología , Estado Nutricional/fisiología , Animales , Hormonas Gastrointestinales/química , Humanos , Osteoblastos/química , Osteoblastos/fisiología , Osteoclastos/química , Osteoclastos/fisiología , Transducción de Señal/fisiologíaRESUMEN
Myostatin (GDF8) is a negative regulator of skeletal muscle growth and mice lacking myostatin show a significant increase in muscle mass and bone density compared to normal mice. In order to further define the role of myostatin in regulating bone mass we sought to determine if loss of myostatin function significantly altered the potential for osteogenic differentiation in bone marrow-derived mesenchymal stem cells in vitro and in vivo. We first examined expression of the myostatin receptor, the type IIB activin receptor (AcvrIIB), in bone marrow-derived mesenchymal stem cells (BMSCs) isolated from mouse long bones. This receptor was found to be expressed at high levels in BMSCs, and we were also able to detect AcvrIIB protein in BMSCs in situ using immunofluorescence. BMSCs isolated from myostatin-deficient mice showed increased osteogenic differentiation compared to wild-type mice; however, treatment of BMSCs from myostatin-deficient mice with recombinant myostatin did not attenuate the osteogenic differentiation of these cells. Loading of BMSCs in vitro increased the expression of osteogenic factors such as BMP-2 and IGF-1, but treatment of BMSCs with recombinant myostatin was found to decrease the expression of these factors. We investigated the effects of myostatin loss-of-function on the differentiation of BMSCs in vivo using hindlimb unloading (7-day tail suspension). Unloading caused a greater increase in marrow adipocyte number, and a greater decrease in osteoblast number, in myostatin-deficient mice than in normal mice. These data suggest that the increased osteogenic differentiation of BMSCs from mice lacking myostatin is load-dependent, and that myostatin may alter the mechanosensitivity of BMSCs by suppressing the expression of osteogenic factors during mechanical stimulation. Furthermore, although myostatin deficiency increases muscle mass and bone strength, it does not prevent muscle and bone catabolism with unloading.
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Diferenciación Celular/fisiología , Suspensión Trasera/fisiología , Células Madre Mesenquimatosas/metabolismo , Células Madre Mesenquimatosas/fisiología , Osteogénesis/fisiología , Factor de Crecimiento Transformador beta/deficiencia , Animales , Células de la Médula Ósea/citología , Proteína Morfogenética Ósea 2 , Proteínas Morfogenéticas Óseas/metabolismo , Células Cultivadas , Factor I del Crecimiento Similar a la Insulina/metabolismo , Ratones , Ratones Endogámicos , Miostatina , Células del Estroma/citología , Células del Estroma/metabolismo , Factor de Crecimiento Transformador beta/genética , Factor de Crecimiento Transformador beta/metabolismoRESUMEN
OBJECTIVE: To determine if myostatin deficiency attenuates body fat gain with increased dietary fat intake. METHODS: Normal and myostatin-deficient mice were fed control (8-10 kcal %fat) and high-fat (HF) (45 kcal %fat) diets for a period of 8 weeks, starting at 2 months of age. Body composition, including percent body fat, lean mass, and fat mass, were measured using DXA. Serum adipokines were measured using a Beadlyte assay. RESULTS: Two-factor ANOVA revealed significant treatment x genotype interactions for body fat (g), percent body fat, and serum leptin. The HF diet significantly increased body fat, percent body fat, and serum leptin in normal mice but not in myostatin-deficient mice. CONCLUSION: Loss of myostatin function not only increases muscle mass in animal models but also attenuates the body fat accumulation that usually accompanies an HF diet.
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Composición Corporal , Grasas de la Dieta/administración & dosificación , Factor de Crecimiento Transformador beta/deficiencia , Aumento de Peso , Adiposidad , Animales , Genotipo , Leptina/sangre , Ratones , Ratones Noqueados , Modelos Animales , Músculo Esquelético/patología , Miostatina , Factor de Crecimiento Transformador beta/genéticaRESUMEN
Tbx2 belongs to a family of developmental transcription regulatory factors. We evaluated whether the gap junction protein Connexin43 (Cx43), an important regulator of osteoblast function and bone development, may be a downstream target gene regulated by Tbx2. The Cx43 promoter contains direct repeats of the consensus T-box binding motif, TCACAC, and moreover, Tbx2 and Cx43 show overlapping expression domains in precursors to bone and in osteoblasts. In vitro analysis showed that the Cx43 promoter contains two Tbx2 binding sites, and this binding was dependent on the TCACAC consensus sequence. Transient transfection analysis with a Cx43 promoter-driven lacZ reporter construct revealed negative regulation mediated by these two Tbx2 binding sites in osteoblast-like cells. Thus, downregulation of Tbx2 led to de-repression of wild-type Cx43 promoter activity, whereas a promoter construct with mutated binding sites showed no de-repression. In stably transfected osteosarcoma cells in which expression of the endogenous Tbx2 gene was downregulated with a Tbx2 antisense construct, a marked de-repression of the endogenous Cx43 gene was observed. This was accompanied by a marked increase in the abundance of Cx43 gap junctions and increased functional gap junction-mediated cell-cell communication. Analysis of lacZ expression in transgenic mice containing the mutated Cx43 promoter-driven lacZ construct further suggested de-repression of the Cx43 promoter in limb buds, a region destined to give rise to long bones of the limbs. Taken together, these findings indicate that the promoter of Cx43 is repressible by Tbx2, both in cultured osteoblast-like cells in vitro and likely in the developing embryo.
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Proteínas de Ciclo Celular/genética , Conexina 43/biosíntesis , Regulación hacia Abajo , Osteoblastos/metabolismo , Proteínas de Dominio T Box/genética , Animales , Secuencia de Bases , Proteínas de Ciclo Celular/metabolismo , Línea Celular Tumoral , Conexina 43/genética , Humanos , Ratones , Ratones Transgénicos , Datos de Secuencia Molecular , Células 3T3 NIH , Osteosarcoma/metabolismo , Ratas , Alineación de Secuencia , Proteínas de Dominio T Box/metabolismo , Transcripción Genética , TransfecciónRESUMEN
Leptin is a hormone secreted by adipocytes that can regulate bone mass through a central, neuroendocrine signaling pathway. We tested the hypothesis that the response of bone tissue to altered leptin signaling is not uniform throughout the skeleton, but may vary between different skeletal regions and between cortical and trabecular moieties. We investigated the effects of leptin deficiency on muscle mass and bone architecture in obese, leptin-deficient (ob/ob) mice, and in lean controls. Results indicate that the obese mice weigh approximately twice as much as the lean mice, but the quadriceps muscles of the ob/ob mice are 40% smaller than those of controls. Leptin-deficient mice have significantly shorter femora, lower femoral bone mineral content (BMC), bone mineral density (BMD), cortical thickness, and trabecular bone volume compared to lean mice. Marrow tissue from the femora of ob/ob mice also shows a marked increase in adipocyte number compared to that of normal mice. In contrast to the pattern observed in the femur, ob/ob mice have significantly increased vertebral length, lumbar BMC, lumbar BMD, and trabecular bone volume compared to lean controls. Few adipocytes are observed in bone marrow from lumbar vertebrae of ob/ob mice, despite being numerous in marrow of the femur. However, like the femur, significant cortical thinning is also observed in the spine. These results indicate that the effects of altered leptin signaling on bone differ significantly between axial and appendicular regions, and may be mediated in part by muscle mass. The muscle hypoplasia, increased marrow adipogenesis, and decreased bone mass observed in the hindlimbs of ob/ob mice are also observed with aging in humans, suggesting that the ob/ob mouse may be a new and useful animal model for studying the relationship between bone marrow adipogenesis and osteopenia.
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Densidad Ósea/genética , Fémur/metabolismo , Leptina/deficiencia , Leptina/genética , Fenotipo , Columna Vertebral/metabolismo , Animales , Fémur/patología , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Obesos , Columna Vertebral/patologíaRESUMEN
Tbx2 is a member of the developmentally important transcriptional regulatory T-box gene family, whose target genes have not been well characterized. In an attempt to identify genes that may be regulated by Tbx2, mouse cDNA microarrays were used to analyze differential gene expression profiles, comparing stably transfected NIH3T3 cells overexpressing Tbx2 and vector-transfected controls. Among 8734 genes, 107 genes were up-regulated by 2-fold or greater, and 66 genes were down-regulated by 2-fold or greater. Caveolin, pleiotrophin (osf-1), osteoblast-specific factor-2 (osf-2) and collagen type I alpha were among the genes upregulated in the Tbx2-overexpressing cells, whereas cadherin 3, tenascin C, and insulin-like growth factor binding protein 10/CYR61 (IBP10) were among the genes downregulated. Northern blot analysis confirmed the correlation of expression of several genes, including IBP10 and osf-2, in fibroblast NIH3T3 and rat osteosarcoma ROS17/2.8 cells differentially expressing Tbx2. In ROS17/2.8 cells transfected with antisense Tbx2, osf-2 was downregulated, whereas transfection of sense Tbx2 upregulated this gene. Interestingly, the expression of pleiotrophin (osf-1) and collagen I alpha with Tbx2 transfection showed an inverse regulatory correlation between NIH3T3 and ROS17/2.8 cells. Thus, Tbx2 can act as both a repressor and activator, and the cellular context can influence the effect on gene expression. Although the data do not address whether Tbx2 directly mediates the transcriptional effect, a number of candidate genes possess putative T-box gene regulatory elements. The results support the hypothesis that Tbx2 may be an important modulator of bone development. Further functional cluster analysis indicates that Tbx2 might also be involved in the regulation of cell cycle and cell adhesion.
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Perfilación de la Expresión Génica , Análisis de Secuencia por Matrices de Oligonucleótidos , Proteínas de Dominio T Box/farmacología , Células 3T3 , Animales , Caveolinas/efectos de los fármacos , Caveolinas/metabolismo , Adhesión Celular/genética , Movimiento Celular/genética , Condrocitos/metabolismo , Bases de Datos Factuales , Regulación de la Expresión Génica/efectos de los fármacos , Humanos , Ratones , Morfogénesis/genética , Osteoblastos/metabolismo , Osteogénesis/genética , Ratas , Proteínas de Dominio T Box/genética , Transfección , Células Tumorales CultivadasRESUMEN
Glucose-dependent insulinotropic peptide (GIP) is a gut-derived hormone known to be important in modulating glucose-induced insulin secretion. In addition, GIP receptors are widely distributed and may have effects on multiple other tissues: fat cells, adrenal glands, endothelium and brain. We have demonstrated recently that GIP also has anabolic effects on bone-derived cells. We now demonstrate that GIP administration prevents the bone loss associated with ovariectomy. We propose that GIP plays a unique role in signaling the bone about nutrient availability, indicating the importance of the gut hormones in directing absorbed nutrients to the bone, and suggesting the concept of an 'entero-osseous axis'. Thus, GIP plays an integrative role helping coordinate efficient and targeted nutrient absorption and distribution.
Asunto(s)
Densidad Ósea/efectos de los fármacos , Polipéptido Inhibidor Gástrico/farmacología , Animales , Desarrollo Óseo/efectos de los fármacos , Modelos Animales de Enfermedad , Femenino , Polipéptido Inhibidor Gástrico/administración & dosificación , Humanos , Immunoblotting , Hibridación in Situ , Masculino , Osteoporosis/tratamiento farmacológico , Ovariectomía , ARN Mensajero/metabolismo , Ratas , Ratas Sprague-Dawley , Receptores de la Hormona Gastrointestinal/genética , Receptores de la Hormona Gastrointestinal/metabolismo , Células Tumorales CultivadasRESUMEN
We have previously shown that angiotensin II (AngII) is able to prime, or sensitize, the secretory response of cultured bovine adrenal glomerulosa cells to the Ca(2+) channel agonist, BAY K8644. We examined the ability of AngII to prime glomerulosa cells to an elevated extracellular K(+) level, a physiological agonist that also triggers Ca(2+) influx. K(+) (9 mM) elicited enhanced secretion in AngII-primed cells compared to those with no prior exposure to the hormone, suggesting that AngII can sensitize glomerulosa cells to respond to increases in extracellular K(+). The potential involvement of protein kinase C (PKC) in priming was investigated by determining whether enhanced Ca(2+) influx could maintain the AngII-induced phosphorylation of the endogenous PKC substrate, myristoylated, alanine-rich C kinase substrate (MARCKS). Incubation with the AngII antagonist, saralasin, for 30 min following an AngII exposure reduced the AngII-induced increase in MARCKS phosphorylation. 100 nM BAY K8644 together with saralasin was unable to maintain AngII-stimulated MARCKS phosphorylation. On the other hand, phosphorylation of the steroidogenic acute regulatory (StAR) protein was sustained with saralasin exposure, both in the presence and absence of BAY K8644. This observation suggests that persistent StAR phosphorylation/activation may play a role in priming.
Asunto(s)
Aldosterona/metabolismo , Angiotensina II/farmacología , Calcio/metabolismo , Péptidos y Proteínas de Señalización Intracelular , Proteínas de la Membrana , Ácido 3-piridinacarboxílico, 1,4-dihidro-2,6-dimetil-5-nitro-4-(2-(trifluorometil)fenil)-, Éster Metílico/farmacología , Angiotensina II/fisiología , Animales , Transporte Biológico/efectos de los fármacos , Agonistas de los Canales de Calcio/farmacología , Bovinos , Sustrato de la Proteína Quinasa C Rico en Alanina Miristoilada , Fosforilación/efectos de los fármacos , Potasio/farmacología , Proteína Quinasa C/antagonistas & inhibidores , Proteína Quinasa C/metabolismo , Proteína Quinasa C/farmacología , Proteínas/metabolismo , Zona Glomerular/citologíaRESUMEN
To generate functional fluorescently tagged glucose-dependent insulinotropic polypeptide (GIP), a series of GIP expression constructs were devised. These included G1 (complete preprohormone), G2 (lacking the C-terminal extension), G3 (lacking both N- and C-terminal extensions), G4 (G2 fused to green fluorescent protein, GFP), and G5 (G3 fused to GFP). Expression of G5 in bacteria generated immunopositive GIP together with GFP fluorescence, while G4 generated only fluorescence without immunoreactivity. Transfection of NIH3T3 cells with cDNAs of G1, G3, G5, but not G2, G4, and EGFP, resulted in immunologically detectable GIP formation, although fluorescence could be detected in the latter two. GIP as well as GIP-GFP secreted by NIH3T3 cells significantly stimulated intracellular cAMP accumulation and Ca(2+) mobilization in SaOS2 cells. The GIP receptor antagonist GIP(7-30) abolished these responses. These results suggest that a GIP-GFP fusion protein seven times larger than the native peptide retains function and may be used as an in vivo probe to detect GIP receptor distribution and to explore GIP's biological roles.
Asunto(s)
Polipéptido Inhibidor Gástrico/metabolismo , Células 3T3 , Animales , Calcio/metabolismo , Cartilla de ADN , ADN Complementario , Proteínas Fluorescentes Verdes , Humanos , Proteínas Luminiscentes/metabolismo , Ratones , Células Tumorales CultivadasRESUMEN
The steroidogenic acute regulatory (StAR) protein mediates cholesterol transport within the mitochondria, and its phosphorylation is believed to be required for steroidogenesis. Increased extracellular potassium concentrations (K(+)), angiotensin II (AngII), and adrenocorticotropic hormone (ACTH) induce aldosterone secretion from bovine adrenal glomerulosa cells. We hypothesized that, although these agonists act via different signaling pathways, StAR phosphorylation should be common to their action. We studied the effects of K(+), AngII, and ACTH, at concentrations that yield comparable secretory responses, on StAR phosphorylation. All three agents induced significant increases in StAR phosphorylation although the response to ACTH was less than that of AngII and K(+). In cells stimulated with the protein kinase C (PKC) agonist 12-tetradecanoylphorbol 13-acetate (TPA), the Ca(2+) channel agonist BAY K8644, and the adenylate cyclase agonist forskolin, TPA caused a small but statistically significant increase in StAR phosphorylation while BAY K8644 and forskolin had no significant effect. Interestingly, the combination of TPA and BAY K8644 produced a larger increase in StAR phosphorylation than the agents alone. We conclude that in cultured bovine adrenal glomerulosa cells the PKC signaling pathway is most effective at inducing StAR phosphorylation but that there is no simple correlation between this event and aldosterone production.
Asunto(s)
Aldosterona/agonistas , Aldosterona/metabolismo , Fosfoproteínas/metabolismo , Ácido 3-piridinacarboxílico, 1,4-dihidro-2,6-dimetil-5-nitro-4-(2-(trifluorometil)fenil)-, Éster Metílico/farmacología , Hormona Adrenocorticotrópica/farmacología , Angiotensina II/farmacología , Animales , Western Blotting , Agonistas de los Canales de Calcio/farmacología , Bovinos , Células Cultivadas , Colforsina/farmacología , Electroforesis en Gel Bidimensional , Fosforilación/efectos de los fármacos , Cloruro de Potasio/farmacología , Proteína Quinasa C/metabolismo , Transducción de Señal/efectos de los fármacos , Acetato de Tetradecanoilforbol/farmacología , Vasoconstrictores/farmacología , Zona Glomerular/citología , Zona Glomerular/efectos de los fármacos , Zona Glomerular/metabolismoRESUMEN
Glucose-dependent insulinotropic peptide (GIP) potentiates glucose-induced insulin secretion. In addition, GIP has vasoconstrictive or vasodilatory properties depending on the vascular bed affected. In order to assess whether this effect could be related to differences in GIP receptor expression, several different endothelial cell types were examined for GIP receptor expression. GIP receptor splice variants were detected and varied depending on the endothelial cell type. Furthermore, stimulation of these cells with GIP led to cell type dependent differences in activation of the calcium and cAMP signaling pathways. To our knowledge this is the first physiological characterization of receptors for GIP in endothelial cells.