RESUMO
Nesfatin-1, an 82 amino acid peptide cleaved from the N-terminal of its precursor nucleobindin-2 (NUCB2), is emerging as a multifunctional peptide in fish. The present study aimed to determine whether nesfatin-1 plays a role in fish somatic growth by modulating the growth hormone (GH)/insulin-like growth factor (IGF) axis, using a representative teleost model, the goldfish (Carassius auratus). The results demonstrated that a single i.p. injection of synthetic goldfish nesfatin-1 significantly decreased the expression of hypothalamic pacap (approximately 90%) and pituitary Gh (approximately 90%) mRNAs at 15 minutes post-injection. Serum GH levels were also reduced as a result of nesfatin-1 administration, by approximately 45% and 55% at 15 and 30 minutes post-injection, respectively. Likewise, in vitro treatment of goldfish dispersed pituitary cells with nesfatin-1 reduced Gh secretion, suggesting that nesfatin-1 acts directly on pituitary somatotrophs to inhibit Gh release. Exposure of cultured liver fragments to nesfatin-1 (0.1, 1 and 10 nmol L-1 ) led to a significant reduction in igf-1 mRNA at 120 minutes and of igf-II mRNA at 30 and 60 minutes post-incubation. Collectively, these results indicate a suppressive role for nesfatin-1 on the goldfish GH/IGF axis. Immunohistochemical studies demonstrated that NUCB2/nesfatin-1-like immunoreactivity, although present in the goldfish pituitary, is not colocalised with GH in goldfish somatotrophs. Thus, nesfatin-1 does not appear to act in an autocrine manner to regulate GH secretion. Taken together, this research found that the pituitary gland is an important source of endogenous NUCB2/nesfatin-1 and also that nesfatin-1 directly suppresses the Gh/IGF axis in goldfish.
Assuntos
Hormônio do Crescimento/antagonistas & inibidores , Nucleobindinas/farmacologia , Somatomedinas/antagonistas & inibidores , Animais , Células Cultivadas , Feminino , Expressão Gênica/efeitos dos fármacos , Carpa Dourada , Hormônio do Crescimento/metabolismo , Fator de Crescimento Insulin-Like I/efeitos dos fármacos , Fator de Crescimento Insulin-Like I/genética , Fator de Crescimento Insulin-Like I/metabolismo , Fator de Crescimento Insulin-Like II/efeitos dos fármacos , Fator de Crescimento Insulin-Like II/genética , Fator de Crescimento Insulin-Like II/metabolismo , Masculino , Nucleobindinas/metabolismo , Hipófise/efeitos dos fármacos , Hipófise/metabolismo , Transdução de Sinais/efeitos dos fármacos , Transdução de Sinais/genética , Somatomedinas/metabolismoRESUMO
Although dioxins and related chemicals have been suspected to disrupt child development, their toxic mechanism remains poorly understood. Our previous studies in rat fetuses revealed that maternal exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), a highly-toxic dioxin, suppresses fetal synthesis of pituitary growth hormone (GH) that is essential for development. This study examined the hypothesis that attenuating GH expression in fetuses triggers developmental disorders. Treating pregnant rats with 1 µg/kg TCDD reduced the circulating level of GH and its downstream factor, insulin-like growth factor-1 (IGF-1), in the offspring only during the fetal and early neonatal stages. Although maternal TCDD exposure resulted in low body weight and length at babyhood and defects in the learning and memory ability at adulthood, GH supplementation in TCDD-exposed fetuses restored or tended to restore the defects including IGF-1 downregulation. Moreover, maternal TCDD exposure decreased the number of GH-positive cells during the fetal/neonatal stage. A microarray analysis showed that TCDD reduced the expression of death-associated protein-like 1 (DAPL1), a cell cycle-dependent proliferation regulator, in the fetal pituitary gland. In addition, TCDD treatment attenuated proliferating cells and cyclin mRNA expression in the fetal pituitary gland. Aryl hydrocarbon receptor (AHR)-knockout fetuses were insensitive to TCDD treatment, indicating that the TCDD-induced reduction in DAPL1 and GH mRNAs expression was due to AHR activation. Finally, DAPL1 knockdown suppressed GH and cyclin D2 expression in fetal pituitary cells. These results provide a novel evidence that dioxin suppresses GH-producing cell proliferation and GH synthesis due to partly targeting DAPL1, thereby impairing offspring development.
Assuntos
Deficiências do Desenvolvimento/metabolismo , Dioxinas/toxicidade , Feto/metabolismo , Hormônio do Crescimento/deficiência , Efeitos Tardios da Exposição Pré-Natal/metabolismo , Animais , Deficiências do Desenvolvimento/induzido quimicamente , Feminino , Feto/efeitos dos fármacos , Hormônio do Crescimento/antagonistas & inibidores , Masculino , Gravidez , Efeitos Tardios da Exposição Pré-Natal/induzido quimicamente , Ratos , Ratos Transgênicos , Ratos WistarRESUMO
OBJECTIVE: Insulin-like growth factor 1 (IGFI) is one of several growth factors which is induced by growth hormone (GH), which activates the Janus kinase 2 (JAK2)-signal transducer and activator of transcription 5 (STAT5) pathway, and plays crucial roles in normal human growth, metabolism, and systemic energy homeostasis. However, little is known about the negative regulation of IGF-I production under different physiological or pathological conditions. Herein, we explore whether activation of endoplasmic reticulum (ER) stress regulates IGF-I production and normal body growth. MATERIALS AND METHODS: C57BL/6 J mice were challenged with tunicamycin (Tm) to induce ER stress activation. 24 h after stimulation, hepatic mRNA expression was analyzed by RNA-Seq and validated by qPCR. Enzyme-linked immunosorbent assay (ELISA) was performed 24 h after Tm stimulation. Body growth was determined 16 days after Tm stimulation. Animals were then sacrificed and liver tissues were collected for further analysis. RESULTS: Mice challenged with Tm displayed a retardation of growth. Molecularly, we found that ER stress inhibited phosphorylation of STAT5. IGF-I transcription and circulating IGF-I were also dramatically decreased under ER stress activation. Moreover, our results demonstrate that IGF-I administration ameliorates Tm-induced growth retardation. CONCLUSIONS: ER stress induces growth retardation. ER stress inhibits hepatic GH-JAK2 signaling activation and its downstream target gene expression. These results warrant further research to explore the crosstalk between ER stress and growth hormone signaling in improving body growth.
Assuntos
Estresse do Retículo Endoplasmático , Transtornos do Crescimento/patologia , Hormônio do Crescimento/antagonistas & inibidores , Fator de Crescimento Insulin-Like I/antagonistas & inibidores , Animais , Transtornos do Crescimento/etiologia , Transtornos do Crescimento/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BLRESUMO
Porcine circovirus type 2 (PCV2) is an important pathogen of the current pig industry. The Wnt signalling pathway plays an important role in the growth of young animals. In this study, we mainly elucidated the relationship between PCV2 and the Wnt signalling pathway. In an in vivo experiment in mice, we demonstrated the downregulatory effects of PCV2 infection on expression levels of downstream components of the Wnt signalling pathway. Weight loss in mice was reversed by activating the Wnt signalling pathway, and the body weight was still significantly higher than that in mice infected with PCV2. We detected levels of growth hormone (GH) in the liver and sera, which showed that GH was also downregulated in mice challenged with PCV2. Lithium chloride, the activator of Wnt signalling, upregulated GH, albeit to a significantly lesser degree than that in corresponding non-stimulated mock mice. In vitro studies showed that PCV2 infection downregulated protein expression of ß-catenin and mRNA expression of matrix metallopeptidase-2 (Mmp2), downregulated protein expression of ß-catenin in the cytoplasm and nucleus, and reduced the activity of the TCF/LEF promoter, demonstrating that PCV2 inhibited activation of the Wnt signalling pathway in vitro. Finally, we found that Rep protein of PCV2 might be responsible for the inhibitory effect.
Assuntos
Circovirus/patogenicidade , Via de Sinalização Wnt , Animais , Linhagem Celular , Núcleo Celular/metabolismo , Núcleo Celular/virologia , Circovirus/classificação , Citoplasma/metabolismo , Citoplasma/virologia , Feminino , Hormônio do Crescimento/antagonistas & inibidores , Hormônio do Crescimento/genética , Metaloproteinase 2 da Matriz/genética , Camundongos , Camundongos Endogâmicos ICR , Suínos , Ativação Transcricional , Redução de Peso , beta Catenina/genéticaRESUMO
The identification of conserved genes and pathways that regulate lifespan but also healthspan has resulted in an improved understanding of the link between nutrients, signal transduction proteins, and aging but has also provided evidence for the existence of multiple "longevity programs," which are selected based on the availability of nutrients. Periodic fasting and other dietary restrictions can promote entry into a long-lasting longevity program characterized by cellular protection and optimal function but can also activate regenerative processes that lead to rejuvenation, which are independent of the aging rate preceding the restricted period. Thus, a "juventology"-based strategy can complement the traditional gerontology approach by focusing not on aging but on the longevity program affecting the life history period in which mortality is very low and organisms remain youthful, healthy, and fully functional.
Assuntos
Restrição Calórica , Dieta com Restrição de Proteínas , Jejum/fisiologia , Expectativa de Vida , Longevidade/genética , Rejuvenescimento/fisiologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Animais , Proteínas Quinases Dependentes de AMP Cíclico/antagonistas & inibidores , Radicais Livres/efeitos adversos , Hormônio do Crescimento/antagonistas & inibidores , Humanos , Fator de Crescimento Insulin-Like I/antagonistas & inibidores , Camundongos , Pessoa de Meia-Idade , Nutrientes/fisiologia , Proteínas Quinases S6 Ribossômicas/antagonistas & inibidores , Saccharomyces cerevisiae/metabolismo , Sirolimo/farmacologia , Serina-Treonina Quinases TOR/antagonistas & inibidores , Adulto JovemRESUMO
OBJECTIVES: Extremely low gestational age neonates with chronic lung disease requiring oxygen therapy frequently experience fluctuations in arterial oxygen saturation or intermittent hypoxia (IH). These infants are at risk for multi-organ developmental delay, reduced growth, and short stature. The growth hormone (GH)/insulin-like growth factor-I (IGF-1) system, an important hormonal regulator of lipid and carbohydrate metabolism, promotes neonatal growth and development. We tested the hypothesis that increasing episodes of IH delay neonatal growth by influencing the GH/IGF-I axis. DESIGN: Newborn rats were exposed to 2, 4, 6, 8, 10, or 12 hypoxic episodes (12% O2) during hyperoxia (50% O2) from P0-P7, P0-P14 (IH), or allowed to recover from P7-P21 or P14-P21 (IHR) in room air (RA). RA littermates at P7, P14, and P21 served as RA controls; and groups exposed to hyperoxia only (50% O2) served as zero IH controls. Histopathology of the liver; hepatic levels of GH, GHBP, IGF-I, IGFBP-3, and leptin; and immunoreactivities of GH, GHR, IGF-I and IGF-IR were determined. RESULTS: Pathological findings of the liver, including cellular swelling, steatosis, necrosis and focal sinusoid congestion were seen in IH, and were particularly severe in the P7 animals. Hepatic GH levels were significantly suppressed in the IH groups exposed to 6-12 hypoxic episodes per day and were not normalized during IHR. Deficits in the GH levels were associated with reduced body length and increase body weight during IHR suggesting increased adiposity and catchup fat. Catchup fat was also associated with elevations in GHBP, IGF-I, leptin. CONCLUSIONS: IH significantly impairs hepatic GH/IGF-1 signaling during the first few weeks of life, which is likely responsible for hepatic GH resistance, increased body fat, and hepatic steatosis. These hormonal perturbations may contribute to long-term organ and body growth impairment, and metabolic dysfunction in preterm infants experiencing frequent IH and/or apneic episodes.
Assuntos
Transtornos do Crescimento/etiologia , Hormônio do Crescimento/antagonistas & inibidores , Hipóxia/complicações , Fator de Crescimento Insulin-Like I/antagonistas & inibidores , Fígado/metabolismo , Animais , Animais Recém-Nascidos , Feminino , Regulação da Expressão Gênica , Transtornos do Crescimento/metabolismo , Transtornos do Crescimento/patologia , Fígado/efeitos dos fármacos , Fígado/patologia , Ratos , Ratos Sprague-DawleyRESUMO
AIM: We studied externally controlled anticancer effects of binding tumor growth inhibiting synthetic peptides to magnetoelectric nanoparticles (MENs) on treatment of glioblastomas. METHODS: Hydrothermally synthesized 30-nm MENs had the core-shell composition of CoFe2O4@BaTiO3. Molecules of growth hormone-releasing hormone antagonist of the MIA class (MIA690) were chemically bound to MENs. In vitro experiments utilized human glioblastoma cells (U-87MG) and human brain microvascular endothelial cells. RESULTS: The studies demonstrated externally controlled high-efficacy binding of MIA690 to MENs, targeted specificity to glioblastoma cells and on-demand release of the peptide by application of d.c. and a.c. magnetic fields, respectively. CONCLUSION: The results support the use of MENs as an effective drug delivery carrier for growth hormone-releasing hormone antagonists in the treatment of human glioblastomas.
Assuntos
Antineoplásicos/química , Neoplasias Encefálicas/tratamento farmacológico , Portadores de Fármacos/química , Glioblastoma/tratamento farmacológico , Hormônio do Crescimento/antagonistas & inibidores , Nanopartículas de Magnetita/química , Peptídeos/química , Antineoplásicos/administração & dosagem , Compostos de Bário/química , Encéfalo/irrigação sanguínea , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Cobalto/química , Liberação Controlada de Fármacos , Células Endoteliais/citologia , Células Endoteliais/efeitos dos fármacos , Compostos Férricos/química , Hormônio do Crescimento/metabolismo , Antagonistas de Hormônios/uso terapêutico , Humanos , Campos Magnéticos , Microvasos/citologia , Nanosferas/química , Tamanho da Partícula , Peptídeos/administração & dosagem , Titânio/químicaRESUMO
The endocrine system is crucial for maintaining whole-body homeostasis. Little is known regarding endocrine hormones secreted by the heart other than atrial/brain natriuretic peptides discovered over 30 years ago. Here, we identify growth differentiation factor 15 (GDF15) as a heart-derived hormone that regulates body growth. We show that pediatric heart disease induces GDF15 synthesis and secretion by cardiomyocytes. Circulating GDF15 in turn acts on the liver to inhibit growth hormone (GH) signaling and body growth. We demonstrate that blocking cardiomyocyte production of GDF15 normalizes circulating GDF15 level and restores liver GH signaling, establishing GDF15 as a bona fide heart-derived hormone that regulates pediatric body growth. Importantly, plasma GDF15 is further increased in children with concomitant heart disease and failure to thrive (FTT). Together these studies reveal a new endocrine mechanism by which the heart coordinates cardiac function and body growth. Our results also provide a potential mechanism for the well-established clinical observation that children with heart diseases often develop FTT.
Assuntos
Desenvolvimento Infantil , Fator 15 de Diferenciação de Crescimento/metabolismo , Hormônio do Crescimento/antagonistas & inibidores , Miócitos Cardíacos/metabolismo , Animais , Pré-Escolar , Humanos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Modelos Animais , Transdução de SinaisRESUMO
In our preliminary studies, we observed zolmitriptan (ZOL) treatment led to induction of CYP3A2 in male not female rats. To figure out the reason is of great significance for drug-drug interactions and personalized administration. Since growth hormone (GH) is known as the major mechanistic determinant of sexually-dimorphic gene expression like CYP3A2 in rat liver, the impacts of ZOL on both plasma GH levels in non monosodium glutamate (MSG)-treated rats and CYP3A2 expression in GH depleted MSG-treated rats were studied. ZOL was shown to partially suppress GH levels in both genders. Furthermore, CYP3A2 protein and mRNA level declined in male not female MSG-treated rats. In order to study the possible molecular events involved in the depression of GH and gender-selective induction on rat CYP3A2 by ZOL, the mRNA and protein level(whole protein and nuclear protein) of hepatocyte nuclear factor 4α (HNF4α) was investigated. Nuclear accumulation of HNF4α was observed in the normal male not female rat liver tissue following ZOL treatment. However, this kind of nuclear translocation did not occur in rat hepatocytes and MSG-treated rats. These findings demonstrated CYP3A2 inducibility by ZOL was gender-selective. GH and HNF4α may play an important role in CYP3A2 induction.
Assuntos
Indutores do Citocromo P-450 CYP3A/farmacologia , Citocromo P-450 CYP3A/metabolismo , Hormônio do Crescimento/antagonistas & inibidores , Oxazolidinonas/farmacologia , Fatores Sexuais , Triptaminas/farmacologia , Animais , Fator 4 Nuclear de Hepatócito/metabolismo , Masculino , RNA Mensageiro , Ratos , Glutamato de SódioRESUMO
Endocrine disruptor chemicals (EDCs) potentially pose a hazard to endangered species. Evaluation of the sensitivity of these species to EDCs could be helpful for protecting their populations. So, the present study investigated the adverse effects of nonylphenol, an EDC, on the endocrine hormones and histopathology of male and female juvenile Caspian brown trout (Salmo trutta caspius) following 21 days of exposure to nominal concentrations of 1, 10 and 100 µg/l. The results showed that the HSI and plasma total calcium of male and female fishes exposed to 100 µg/l nonylphenol were significantly increased compared with the control groups (P<0.001). The male plasma T3 level was significantly decreased in 10 (P<0.01) and 100 (P<0.001) µg/l nonylphenol. The female T3 level increased in 1 µg/l nonylphenol concentration (P<0.05). The plasma T4 of males showed significant elevation in fishes exposed to 100 µg/l nonylphenol (P<0.05), but no change for females in any of treatment groups relative to controls (P>0.05). No significant effect of nonylphenol exposure was observed on male plasma TSH levels (P>0.05), whereas, in females, nonylphenol at all concentrations significantly reduced TSH levels. A bell-shaped response was observed in male and female plasma GH levels. Moreover, various histopathological lesions were observed in gill and intestine tissues of fishes exposed to different nonylphenol concentrations. These results demonstrate the high sensitivity of this endangered species to even environmentally relevant concentrations of nonylphenol. Furthermore, Caspian brown trout could be used as bioindicators reflecting the toxicity of nonylphenol.
Assuntos
Disruptores Endócrinos/toxicidade , Brânquias/efeitos dos fármacos , Intestinos/efeitos dos fármacos , Fenóis/toxicidade , Glândula Tireoide/efeitos dos fármacos , Truta/fisiologia , Poluentes Químicos da Água/toxicidade , Animais , Aquicultura , Biomarcadores/sangue , Cálcio/sangue , Cálcio/química , Relação Dose-Resposta a Droga , Disruptores Endócrinos/administração & dosagem , Disruptores Endócrinos/sangue , Feminino , Proteínas de Peixes/agonistas , Proteínas de Peixes/antagonistas & inibidores , Proteínas de Peixes/sangue , Proteínas de Peixes/metabolismo , Brânquias/crescimento & desenvolvimento , Brânquias/patologia , Hormônio do Crescimento/agonistas , Hormônio do Crescimento/antagonistas & inibidores , Hormônio do Crescimento/sangue , Hormônio do Crescimento/metabolismo , Intestinos/crescimento & desenvolvimento , Intestinos/patologia , Masculino , Fenóis/administração & dosagem , Fenóis/sangue , Distribuição Aleatória , Caracteres Sexuais , Glândula Tireoide/crescimento & desenvolvimento , Glândula Tireoide/metabolismo , Hormônios Tireóideos/agonistas , Hormônios Tireóideos/sangue , Hormônios Tireóideos/química , Hormônios Tireóideos/metabolismo , Tireotropina/antagonistas & inibidores , Tireotropina/sangue , Tireotropina/metabolismo , Testes de Toxicidade Subaguda/métodos , Toxicocinética , Truta/sangue , Truta/crescimento & desenvolvimento , Truta/metabolismo , Poluentes Químicos da Água/administração & dosagem , Poluentes Químicos da Água/sangueRESUMO
After significant injury, the liver must maintain homeostasis during the regenerative process. We hypothesized the existence of mechanisms to limit hepatocyte proliferation after injury to maintain metabolic and synthetic function. A screen for candidates revealed suppressor of cytokine signaling 2 (SOCS2), an inhibitor of growth hormone (GH) signaling, was strongly induced after partial hepatectomy. Using genetic deletion and administration of various factors we investigated the role of SOCS2 during liver regeneration. SOCS2 preserves liver function by restraining the first round of hepatocyte proliferation after partial hepatectomy by preventing increases in growth hormone receptor (GHR) via ubiquitination, suppressing GH pathway activity. At later times, SOCS2 enhances hepatocyte proliferation by modulating a decrease in serum insulin-like growth factor 1 (IGF-1) that allows GH release from the pituitary. SOCS2, therefore, plays a dual role in modulating the rate of hepatocyte proliferation. In particular, this is the first demonstration of an endogenous mechanism to limit hepatocyte proliferation after injury.
Assuntos
Fator de Crescimento Insulin-Like I/antagonistas & inibidores , Regeneração Hepática , Fígado/fisiologia , Receptores da Somatotropina/antagonistas & inibidores , Proteínas Supressoras da Sinalização de Citocina/metabolismo , Ubiquitinação , Animais , Proliferação de Células , Células Cultivadas , Regulação da Expressão Gênica , Hormônio do Crescimento/antagonistas & inibidores , Hormônio do Crescimento/metabolismo , Hepatectomia/efeitos adversos , Imuno-Histoquímica , Fator de Crescimento Insulin-Like I/análise , Fígado/citologia , Fígado/cirurgia , Masculino , Camundongos Endogâmicos C57BL , Camundongos Knockout , Hipófise/citologia , Hipófise/metabolismo , Transporte Proteico , Proteólise , Receptores da Somatotropina/agonistas , Receptores da Somatotropina/genética , Receptores da Somatotropina/metabolismo , Proteínas Supressoras da Sinalização de Citocina/genéticaRESUMO
Protein malnutrition promotes hepatic steatosis, decreases insulin-like growth factor (IGF)-I production and retards growth. To identify new molecules involved in such changes, we conducted DNA microarray analysis on liver samples from rats fed an isoenergetic low-protein diet for 8 h. We identified the fibroblast growth factor 21 gene (Fgf21) as one of the most strongly up-regulated genes under conditions of acute protein malnutrition (P<0·05, false-discovery rate<0·001). In addition, amino acid deprivation increased Fgf21 mRNA levels in rat liver-derived RL-34 cells (P<0·01). These results suggested that amino acid limitation directly increases Fgf21 expression. FGF21 is a polypeptide hormone that regulates glucose and lipid metabolism. FGF21 also promotes a growth hormone-resistance state and suppresses IGF-I in transgenic mice. Therefore, to determine further whether Fgf21 up-regulation causes hepatic steatosis and growth retardation after IGF-I decrease in protein malnutrition, we fed an isoenergetic low-protein diet to Fgf21-knockout (KO) mice. Fgf21-KO did not rescue growth retardation and reduced plasma IGF-I concentration in these mice. Fgf21-KO mice showed greater epididymal white adipose tissue weight and increased hepatic TAG and cholesterol levels under protein malnutrition conditions (P<0·05). Overall, the results showed that protein deprivation directly increased Fgf21 expression. However, growth retardation and decreased IGF-I were not mediated by increased FGF21 expression in protein malnutrition. Furthermore, FGF21 up-regulation rather appears to have a protective effect against obesity and hepatic steatosis in protein-malnourished animals.
Assuntos
Dieta com Restrição de Proteínas , Fatores de Crescimento de Fibroblastos/metabolismo , Metabolismo dos Lipídeos , Desnutrição Proteico-Calórica/metabolismo , Tecido Adiposo Branco/metabolismo , Animais , Colesterol/metabolismo , Fígado Gorduroso/genética , Fatores de Crescimento de Fibroblastos/genética , Hormônio do Crescimento/antagonistas & inibidores , Hormônio do Crescimento/metabolismo , Fator de Crescimento Insulin-Like I/antagonistas & inibidores , Fator de Crescimento Insulin-Like I/genética , Fator de Crescimento Insulin-Like I/metabolismo , Fígado/metabolismo , Masculino , Camundongos , Camundongos Knockout , Obesidade , Análise de Sequência com Séries de Oligonucleotídeos , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Ratos , Ratos Wistar , Triglicerídeos/metabolismo , Regulação para CimaRESUMO
Chronic exposure to trichothecenes is known to disturb insulin-like growth factor 1 and signaling of insulin and leptin hormones and causes considerable growth retardation in animals. However, limited information was available on mechanisms underlying trichothecene-induced growth retardation. In this study, we employed an integrated transcriptomics, proteomics, and RNA interference (RNAi) approach to study the molecular mechanisms underlying trichothecene cytotoxicity in rat pituitary adenoma GH3 cells. Our results showed that trichothecenes suppressed the synthesis of growth hormone 1 (Gh1) and inhibited the eukaryotic transcription and translation initiation by suppressing aminoacyl-tRNA synthetases transcription, inducing eukaryotic translation initiation factor 2-alpha kinase 2 (EIF2AK2) and reducing eukaryotic translation initiation factor 5 a. The sulfhydryl oxidases , protein disulfide isomerase,and heat shock protein 90 (were greatly reduced, which resulted in adverse regulation of protein processing and folding. Differential genes and proteins associated with a decline in energy metabolism and cell cycle arrest were also found in our study. However, use of RNAi to interfere with hemopoietic cell kinase (Hck) and EIF2AK2 transcriptions or use of chemical inhibitors of MAPK, p38, Ras, and JNK partially reversed the reduction of Gh1 levels induced by trichothecenes. It indicated that the activation of MAPKs, Hck, and EIF2AK2 were important for trichothecene-induced growth hormone suppression. Considering the potential hazards of exposure to trichothecenes, our findings could help to improve our understanding regarding human and animal health implications.
Assuntos
Perfilação da Expressão Gênica , Hormônio do Crescimento/antagonistas & inibidores , Proteômica , Toxina T-2/análogos & derivados , Aminoacil-tRNA Sintetases/antagonistas & inibidores , Animais , Apoptose/efeitos dos fármacos , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Proteínas de Choque Térmico HSP90/antagonistas & inibidores , Oxirredutases/antagonistas & inibidores , Fatores de Iniciação de Peptídeos/antagonistas & inibidores , Biossíntese de Proteínas/efeitos dos fármacos , Isomerases de Dissulfetos de Proteínas/antagonistas & inibidores , Interferência de RNA/efeitos dos fármacos , Proteínas de Ligação a RNA/antagonistas & inibidores , Ratos , Toxina T-2/farmacologia , Transcrição Gênica/efeitos dos fármacos , eIF-2 Quinase/antagonistas & inibidores , Fator de Iniciação de Tradução Eucariótico 5ARESUMO
The workshop entitled 'Interventions to Slow Aging in Humans: Are We Ready?' was held in Erice, Italy, on October 8-13, 2013, to bring together leading experts in the biology and genetics of aging and obtain a consensus related to the discovery and development of safe interventions to slow aging and increase healthy lifespan in humans. There was consensus that there is sufficient evidence that aging interventions will delay and prevent disease onset for many chronic conditions of adult and old age. Essential pathways have been identified, and behavioral, dietary, and pharmacologic approaches have emerged. Although many gene targets and drugs were discussed and there was not complete consensus about all interventions, the participants selected a subset of the most promising strategies that could be tested in humans for their effects on healthspan. These were: (i) dietary interventions mimicking chronic dietary restriction (periodic fasting mimicking diets, protein restriction, etc.); (ii) drugs that inhibit the growth hormone/IGF-I axis; (iii) drugs that inhibit the mTOR-S6K pathway; or (iv) drugs that activate AMPK or specific sirtuins. These choices were based in part on consistent evidence for the pro-longevity effects and ability of these interventions to prevent or delay multiple age-related diseases and improve healthspan in simple model organisms and rodents and their potential to be safe and effective in extending human healthspan. The authors of this manuscript were speakers and discussants invited to the workshop. The following summary highlights the major points addressed and the conclusions of the meeting.
Assuntos
Envelhecimento/efeitos dos fármacos , Fatores Biológicos/uso terapêutico , Longevidade/efeitos dos fármacos , Medicamentos sob Prescrição/uso terapêutico , Proteínas Quinases Ativadas por AMP/genética , Proteínas Quinases Ativadas por AMP/metabolismo , Envelhecimento/genética , Animais , Restrição Calórica/métodos , Dieta , Ativação Enzimática , Regulação da Expressão Gênica , Hormônio do Crescimento/antagonistas & inibidores , Hormônio do Crescimento/genética , Hormônio do Crescimento/metabolismo , Humanos , Fator de Crescimento Insulin-Like I/antagonistas & inibidores , Fator de Crescimento Insulin-Like I/genética , Fator de Crescimento Insulin-Like I/metabolismo , Longevidade/genética , Camundongos , Proteínas Quinases S6 Ribossômicas/antagonistas & inibidores , Proteínas Quinases S6 Ribossômicas/genética , Proteínas Quinases S6 Ribossômicas/metabolismo , Transdução de Sinais , Sirtuínas/genética , Sirtuínas/metabolismo , Serina-Treonina Quinases TOR/antagonistas & inibidores , Serina-Treonina Quinases TOR/genética , Serina-Treonina Quinases TOR/metabolismoRESUMO
ß-hydroxybutyric acid (BHBA) regulates the synthesis and secretion of growth hormone (GH) and prolactin (PRL), but its mechanism is unknown. In this study, we detected the effects of BHBA on the activities of G protein signaling pathways, AMPK-α activity, GH, and PRL gene transcription, and GH and PRL secretion in dairy cow anterior pituitary cells (DCAPCs). The results showed that BHBA decreased intracellular cAMP levels and a subsequent reduction in protein kinase A (PKA) activity. Inhibition of PKA activity reduced cAMP response element-binding protein (CREB) phosphorylation, thereby inhibiting GH and PRL transcription and secretion. The effects of BHBA were attenuated by a specific Gαi inhibitor, pertussis toxin (PTX). In addition, intracellular BHBA uptake mediated by monocarboxylate transporter 1 (MCT1) could trigger AMPK signaling and result in the decrease in GH and PRL mRNA translation in DCAPCs cultured under low-glucose and non-glucose condition when compared with the high-glucose group. This study identifies a biochemical mechanism for the regulatory action of BHBA on GH and PRL gene transcription, translation, and secretion in DCAPCs, which may be one of the factors that regulate pituitary function during the transition period in dairy cows.
Assuntos
Hormônio do Crescimento/metabolismo , Hidroxibutiratos/toxicidade , Adeno-Hipófise/efeitos dos fármacos , Prolactina/metabolismo , Transdução de Sinais/efeitos dos fármacos , Proteínas Quinases Ativadas por AMP/metabolismo , Animais , Bovinos , Células Cultivadas , AMP Cíclico/metabolismo , Proteína de Ligação ao Elemento de Resposta ao AMP Cíclico/metabolismo , Proteínas Quinases Dependentes de AMP Cíclico/metabolismo , Feminino , Glucose/farmacologia , Hormônio do Crescimento/antagonistas & inibidores , Hormônio do Crescimento/genética , Transportadores de Ácidos Monocarboxílicos/genética , Transportadores de Ácidos Monocarboxílicos/metabolismo , Toxina Pertussis/farmacologia , Fosforilação/efeitos dos fármacos , Adeno-Hipófise/citologia , Adeno-Hipófise/metabolismo , Prolactina/antagonistas & inibidores , RNA Mensageiro/metabolismo , Simportadores/genética , Simportadores/metabolismo , Fator de Transcrição Pit-1/genética , Fator de Transcrição Pit-1/metabolismo , Transcrição Gênica/efeitos dos fármacosRESUMO
The development of new growth hormone (GH) agonists and growth hormone antagonists (GHAs) requires animal models for pre-clinical testing. Ideally, the effects of treatment are monitored using the same pharmacodynamic marker that is later used in clinical practice. However, intact rodents are of limited value for this purpose because serum IGF-I, the most sensitive pharmacodynamic marker for the action of GH in humans, shows no response to treatment with recombinant human GH and there is little evidence for the effects of GHAs, except when administered at very high doses or when overexpressed. As an alternative, more suitable model, we explored pharmacodynamic markers of GH action in intact rabbits. We performed the first validation of an IGF-I assay for the analysis of rabbit serum and tested precision, sensitivity, linearity and recovery using an automated human IGF-I assay (IDS-iSYS). Furthermore, IGF-I was measured in rabbits of different strains, age groups and sexes, and we monitored IGF-I response to treatment with recombinant human GH or the GHA Pegvisomant. For a subset of samples, we used LC-MS/MS to measure IGF-I, and quantitative western ligand blot to analyze IGF-binding proteins (IGFBPs). Although recovery of recombinant rabbit IGF-I was only 50% in the human IGF-I assay, our results show that the sensitivity, precision (1.7-3.3% coefficient of variation) and linearity (90.4-105.6%) were excellent in rabbit samples. As expected, sex, age and genetic background were major determinants of IGF-I concentration in rabbits. IGF-I and IGFBP-2 levels increased after single and multiple injections of recombinant human GH (IGF-I: 286±22 versus 434±26 ng/ml; P<0.01) and were highly correlated (P<0.0001). Treatment with the GHA lowered IGF-I levels from the fourth injection onwards (P<0.01). In summary, we demonstrated that the IDS-iSYS IGF-I immunoassay can be used in rabbits. Similar to rodents, rabbits display variations in IGF-I depending on sex, age and genetic background. Unlike in rodents, the IGF-I response to treatment with recombinant human GH or a GHA closely mimics the pharmacodynamics seen in humans, suggesting that rabbits are a suitable new model to test human GH agonists and antagonists.
Assuntos
Biomarcadores/sangue , Hormônio do Crescimento/agonistas , Hormônio do Crescimento/antagonistas & inibidores , Fator de Crescimento Insulin-Like I/metabolismo , Animais , Cromatografia Líquida , Limite de Detecção , Coelhos , Reprodutibilidade dos Testes , Espectrometria de Massas em TandemRESUMO
BACKGROUND: Accumulating evidence suggests that growth hormone (GH) may play a major role in the regulation of postnatal neurogenesis, thus supporting the possibility that it may be also involved in promoting brain repair after brain injury. In order to gain further insight on this possibility, in this study we have investigated the pathways signaling the effect of GH treatment on the proliferation and survival of hippocampal subgranular zone (SGZ)-derived neurospheres. RESULTS: Our results demonstrate that GH treatment promotes both proliferation and survival of SGZ neurospheres. By using specific chemical inhibitors we have been also able to demonstrate that GH treatment promotes the activation of both Akt-mTOR and JNK signaling pathways, while blockade of these pathways either reduces or abolishes the GH effects. In contrast, no effect of GH on the activation of the Ras-ERK pathway was observed after GH treatment, despite blockade of this signaling path also resulted in a significant reduction of GH effects. Interestingly, SGZ cells were also capable of producing GH, and blockade of endogenous GH also resulted in a decrease in the proliferation and survival of SGZ neurospheres. CONCLUSIONS: Altogether, our findings suggest that GH treatment may promote the proliferation and survival of neural progenitors. This effect may be elicited by cooperating with locally-produced GH in order to increase the response of neural progenitors to adequate stimuli. On this view, the possibility of using GH treatment to promote neurogenesis and cell survival in some acquired neural injuries may be envisaged.
Assuntos
Proliferação de Células/fisiologia , Sobrevivência Celular/fisiologia , Hormônio do Crescimento/metabolismo , Hipocampo/fisiologia , Células-Tronco Neurais/fisiologia , Neurogênese/fisiologia , Animais , Apoptose/efeitos dos fármacos , Apoptose/fisiologia , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , MAP Quinases Reguladas por Sinal Extracelular/antagonistas & inibidores , MAP Quinases Reguladas por Sinal Extracelular/metabolismo , Hormônio do Crescimento/antagonistas & inibidores , Hipocampo/efeitos dos fármacos , Humanos , Proteínas Quinases JNK Ativadas por Mitógeno/antagonistas & inibidores , Proteínas Quinases JNK Ativadas por Mitógeno/metabolismo , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Sistema de Sinalização das MAP Quinases/fisiologia , Camundongos Endogâmicos C57BL , Células-Tronco Neurais/efeitos dos fármacos , Neurogênese/efeitos dos fármacos , Fosfatidilinositol 3-Quinases/metabolismo , Inibidores de Fosfoinositídeo-3 Quinase , Proteínas Proto-Oncogênicas c-akt/antagonistas & inibidores , Proteínas Proto-Oncogênicas c-akt/metabolismo , Proteínas Recombinantes/metabolismo , Serina-Treonina Quinases TOR/antagonistas & inibidores , Serina-Treonina Quinases TOR/metabolismo , Proteínas ras/antagonistas & inibidores , Proteínas ras/metabolismoRESUMO
This study aimed to evaluate the efficacy and safety of an oral DNA vaccine against somatostatin (SS) (pGS/2SS-asd, encoding two copies of somatostatin genes) mediated by attenuated Salmonella choleraesuis C500 without antibiotic resistance gene on piglets growth. A total of 50 piglets were uniformly divided into five groups. The animals in the first three groups were orally given vaccine in dose of either 5 9 1010, 5 9 109 or 5 9 108 colony-forming units (CFU).The remaining two groups were orally administered with either bacteria C500(containing pVAX-asd plasmid without somatostatin gene) or phosphate buffered saline (PBS) as controls. The results indicated that the vaccine induced SS-specific antibodies in a dose-dependent pattern. Compared with the PBS control, animals in the high-dose group showed lower SS levels and higher growth hormone (GH) levels in sera. Average daily gain of animals in the high dose group was increased by 32.88% and 26.46% during 4 and 8 weeks,respectively. Anti-SS antibodies were positively correlated with either GH levels or average daily gain at week 8 after primary immunization (P < 0.05). Faecal,soil and water samples originating from immunized piglets and surrounding environment were collected. The target gene (the fusion gene GS/2SS) of C500(pGS/2SS-asd) was not detected by PCR amplification in these samples,indicating that the surrounding environment was not contaminated by residual recombinant bacteria. In conclusion, the vaccine without antibiotic resistance gene is attributable to improve growth performance of piglets through an influence on GH secretion. Moreover, the immunization did not contaminate the surrounding environment of animals.
Assuntos
Hormônio do Crescimento/metabolismo , Salmonella arizonae/genética , Somatostatina/antagonistas & inibidores , Somatostatina/imunologia , Suínos/crescimento & desenvolvimento , Vacinas de DNA/administração & dosagem , Administração Oral , Animais , Anticorpos Antinucleares/imunologia , Resistência Microbiana a Medicamentos/genética , Vetores Genéticos/genética , Hormônio do Crescimento/antagonistas & inibidores , Hormônio do Crescimento/sangue , Reação em Cadeia da Polimerase , Somatostatina/genética , Vacinação , Vacinas de DNA/efeitos adversos , Vacinas de DNA/genéticaRESUMO
Somatostatin receptors (SSTRs) are proposed to mediate the actions of somatostatin (SST) and its related peptide, cortistatin (CST), in vertebrates. However, the identity, functionality, and tissue expression of these receptors remain largely unknown in most non-mammalian vertebrates including birds. In this study, five SSTRs (named cSSTR1, cSSTR2, cSSTR3, cSSTR4, cSSTR5) were cloned from chicken brain by RT-PCR. Using a pGL3-CRE-luciferase reporter system, we demonstrated that activation of each cSSTR expressed in CHO cells by cSST28, cSST14 and cCST14 treatment could inhibit forskolin-induced luciferase activity of CHO cells, indicating the functional coupling of all cSSTRs to Gi protein(s). Interestingly, cSSTR1-4 expressed in CHO cells could be activated by cSST28, cSST14 and cCST14 with high potencies, suggesting that they may function as the receptors common for these peptides. In contrast, cSSTR5 could be potently activated by cSST28 only, indicating that it is a cSST28-specific receptor. Using RT-PCR, wide expression of cSSTRs was detected in chicken tissues including pituitary. In accordance with their expression in pituitary, cSST28, cSST14, and cCST14 were demonstrated to inhibit basal and novel cGHRH1-27NH2-induced GH secretion in cultured chicken pituitary cells dose-dependently (0-10nM) by Western blot analysis, suggesting the involvement of cSSTR(s) common for these peptides in mediating their inhibitory actions. Collectively, our study establishes a molecular basis to elucidate the roles of SST/CST in birds and provide insights into the roles of SST/CST in vertebrates, such as their conserved actions on pituitary.
Assuntos
Galinhas/genética , Hormônio do Crescimento/metabolismo , Neuropeptídeos/genética , Receptores de Somatostatina/genética , Somatostatina/genética , Somatotrofos/metabolismo , Animais , Células CHO , Células Cultivadas , Galinhas/metabolismo , Cricetulus , Regulação da Expressão Gênica , Genes Reporter , Hormônio do Crescimento/antagonistas & inibidores , Hormônio Liberador de Hormônio do Crescimento/farmacologia , Luciferases/genética , Luciferases/metabolismo , Neuropeptídeos/metabolismo , Neuropeptídeos/farmacologia , Isoformas de Proteínas/genética , Isoformas de Proteínas/metabolismo , Receptores de Somatostatina/metabolismo , Somatostatina/metabolismo , Somatostatina/farmacologia , Somatotrofos/citologia , Somatotrofos/efeitos dos fármacos , TransfecçãoRESUMO
Short-chain fatty acids (SCFAs) play a key role in altering carbohydrate and lipid metabolism, influence endocrine pancreas activity, and as a precursor of ruminant milk fat. However, the effect and detailed mechanisms by which SCFAs mediate bovine growth hormone (GH) and prolactin (PRL) gene transcription remain unclear. In this study, we detected the effects of SCFAs (acetate, propionate, and butyrate) on the activity of the cAMP/PKA/CREB signaling pathway, GH, PRL, and Pit-1 gene transcription in dairy cow anterior pituitary cells (DCAPCs). The results showed that SCFAs decreased intracellular cAMP levels and a subsequent reduction in PKA activity. Inhibition of PKA activity decreased CREB phosphorylation, thereby inhibiting GH and PRL gene transcription. Furthermore, PTX blocked SCFAs- inhibited cAMP/PKA/CREB signaling pathway. These data showed that the inhibition of GH and PRL gene transcription induced by SCFAs is mediated by Gi activation and that propionate is more potent than acetate and butyrate in inhibiting GH and PRL gene transcription. In conclusion, this study identifies a biochemical mechanism for the regulation of SCFAs on bovine GH and PRL gene transcription in DCAPCs, which may serve as one of the factors that regulate pituitary function in accordance with dietary intake.