RESUMO
Estrogen regulates fat mass and distribution and glucose metabolism. We have previously found that estrogen sulfotransferase (EST), which inactivates estrogen through sulfoconjugation, was highly expressed in adipose tissue of male mice and induced by testosterone in female mice. To determine whether inhibition of estrogen in female adipose tissue affects adipose mass and metabolism, we generated transgenic mice expressing EST via the aP2 promoter. As expected, EST expression was increased in adipose tissue as well as macrophages. Parametrial and subcutaneous inguinal adipose mass and adipocyte size were significantly reduced in EST transgenic mice, but there was no change in retroperitoneal or brown adipose tissue. EST overexpression decreased the differentiation of primary adipocytes, and this was associated with reductions in the expression of peroxisome proliferator-activated receptor-γ, fatty acid synthase, hormone-sensitive lipase, lipoprotein lipase, and leptin. Serum leptin levels were significantly lower in EST transgenic mice, whereas total and high-molecular-weight adiponectin levels were not different in transgenic and wild-type mice. Glucose uptake was blunted in parametrial adipose tissue during hyperinsulinemic-euglycemic clamp in EST transgenic mice. In contrast, hepatic insulin sensitivity was improved but muscle insulin sensitivity did not change in EST transgenic mice. These results reveal novel effects of EST on adipose tissue and glucose homeostasis in female mice.
Assuntos
Tecido Adiposo/metabolismo , Composição Corporal/fisiologia , Glucose/metabolismo , Resistência à Insulina/fisiologia , Sulfotransferases/metabolismo , Adipócitos Brancos/enzimologia , Adipócitos Brancos/metabolismo , Tecido Adiposo/citologia , Tecido Adiposo/enzimologia , Animais , Diferenciação Celular/fisiologia , Estrogênios/metabolismo , Ácido Graxo Sintases/sangue , Feminino , Técnica Clamp de Glucose , Leptina/sangue , Lipase Lipoproteica/sangue , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , PPAR gama/sangue , Distribuição Aleatória , Esterol Esterase/sangueRESUMO
CYP2C11, the most commonly expressed isoform of cytochrome P450 in male rat liver, was measured in spleen, thymus and bone marrow by quantitative real-time PCR and enhanced Western blotting. CYP2C11 concentrations in the lymphoid tissues were a fraction of that observed in liver, but like the liver, were sexually dimorphic (M>F) with mRNA and protein levels in agreement. Although the response to hypophysectomy varied according to tissue and sex, expression levels of CYP2C11 in all measured tissues remained greater in males. Further differences in CYP2C11 expression between liver and lymphoid tissue were observed following restoration of the circulating masculine growth hormone profile in hypophysectomized rats. In contrast to the liver where the renaturalized growth hormone profile elevated CYP2C11 expression in both sexes, the response was opposite in spleen and thymus with isoform concentrations declining in both sexes. Lastly, the divergent response of CYP2C11 between the liver and immune system was examined in cultured splenocytes exposed to different mitogens. In contrast to the dramatic depletion of CYP2C11 reported in proliferating hepatocytes, mitogen-stimulation resulted in a significant elevation in splenocyte CYP2C11 expression. In summary, we report for the first time that thymus, spleen and bone marrow express, albeit nominal, sex-dependent levels of CYP2C11 (M>F) whose regulation appears to be under some hormonal control, but very different from that of the hepatic isoform.
Assuntos
Hidrocarboneto de Aril Hidroxilases/metabolismo , Medula Óssea/enzimologia , Hormônio do Crescimento/farmacologia , Baço/enzimologia , Esteroide 16-alfa-Hidroxilase/metabolismo , Timo/enzimologia , Animais , Hidrocarboneto de Aril Hidroxilases/genética , Família 2 do Citocromo P450 , Feminino , Hipofisectomia , Fígado/efeitos dos fármacos , Fígado/enzimologia , Masculino , Mitógenos/farmacologia , RNA Mensageiro/metabolismo , Ratos , Ratos Sprague-Dawley , Caracteres Sexuais , Baço/efeitos dos fármacos , Esteroide 16-alfa-Hidroxilase/genética , Timo/efeitos dos fármacosRESUMO
Inherent sex differences in various parameters of growth, musculoskeletal function, metabolism, and cytochrome P450 (P450)-dependent drug metabolism have been reported in rats and humans administered typical intermittent/episodic growth hormone (GH) replacement therapy. Having infused and monitored the identical physiologic masculine (episodic) growth hormone profile to both hypophysectomized male and female rats, we observed that induction levels of hepatic CYP2C11 were 35 to 40% lower in females. Associated with the reduced expression of the P450 isoform in the episodic GH-treated females were dramatically lower activation levels of Janus kinase (Jak2), signal transducers and activators of transcription (Stat5A and 5B) as well as 50% less binding of Stat5B to the CYP2C11 promoter. Because the Jak2/Stat5B signaling pathway mediates the effects of the masculine GH profile on its target cells, we conclude that the lower induction level of CYP2C11 in females exposed to the masculine GH profile is probably due, at least in part, to the suboptimum activation of the Jak2/Stat5B pathway. In addition to the reduced activation of the Jak2/Stat5B pathway, we observed lower activational levels of mitogen-activated protein kinase (p44/p42) and, indirectly, nuclear factor-kappaB in the episodic GH-treated females that may be involved in attenuating the activity of the Jak2/Stat5B pathway diminishing CYP2C11 expression levels.
Assuntos
Hidrocarboneto de Aril Hidroxilases/metabolismo , Hormônio do Crescimento/farmacologia , Janus Quinase 2/metabolismo , Fator de Transcrição STAT5/metabolismo , Transdução de Sinais/efeitos dos fármacos , Esteroide 16-alfa-Hidroxilase/metabolismo , Animais , Hidrocarboneto de Aril Hidroxilases/genética , Western Blotting , Citocromo P-450 CYP3A , Família 2 do Citocromo P450 , Feminino , Expressão Gênica/efeitos dos fármacos , Hormônio do Crescimento/administração & dosagem , Hormônio do Crescimento/sangue , Hipofisectomia , Proteínas I-kappa B/metabolismo , Fígado/citologia , Fígado/metabolismo , Masculino , Proteínas de Membrana/genética , Proteínas de Membrana/metabolismo , Proteínas Quinases Ativadas por Mitógeno/metabolismo , Inibidor de NF-kappaB alfa , Fosforilação/efeitos dos fármacos , Regiões Promotoras Genéticas/genética , Ligação Proteica , Ratos , Ratos Sprague-Dawley , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Fatores Sexuais , Esteroide 16-alfa-Hidroxilase/genética , Esteroide Hidroxilases/genética , Esteroide Hidroxilases/metabolismoRESUMO
CYP2C11, the most commonly expressed hepatic cytochrome P450 isoform in male rats, is induced by the masculine "episodic" secretory growth hormone profile. A considerable number of reports have indicated that episodic growth hormone effects are mediated by the activation of the Janus kinase 2 (Jak2)/signal transducer and activator of transcription (Stat)5B signal transduction pathway. We observed that restoration of the normal masculine plasma growth hormone pulse in hypophysectomized male rats did indeed rapidly activate (phosphorylate) Jak2, shortly followed by activation and nuclear translocation of Stat5B. Infusion of a growth hormone pulse with an amplitude that was 10% of the normal height induced a dramatic overexpression of CYP2C11, had little effect activating Jak2, but induced a more rapid and greater accumulation of activated nuclear Stat5B. Restoration of a growth hormone pulse with an amplitude of only 1% of normal had little effect phosphorylating Jak2, activated and translocated to the hepatic nucleus approximately 70% of the normally induced levels of Stat5B, but had no inductive effect on CYP2C11. Last, the hypophysectomized male rat receiving no growth hormone replacement expressed 25 to 35% of normal concentrations of CYP2C11 despite no measurable activation of either Jak2 or Stat5B. These results raise concerns regarding the requisite role of the Jak2/Stat5B pathway in mediating episodic growth hormone regulation of CYP2C11. However, accumulation of activated extracellular signal-regulated kinase (ERK)1 and ERK2 were the only transducers measured in the study not affected by the 1% replacement pulse of growth hormone and were elevated 2- to 3-fold above normal when the pulse was renaturalized to 10% of physiological amplitude, suggesting the possible involvement of mitogen-activated protein kinase in episodic growth hormone regulation of CYP2C11.
Assuntos
Hidrocarboneto de Aril Hidroxilases/genética , Proteínas de Ligação a DNA/fisiologia , Regulação Enzimológica da Expressão Gênica/efeitos dos fármacos , Hormônio do Crescimento/farmacologia , Fígado/enzimologia , Proteínas Tirosina Quinases/metabolismo , Proteínas Proto-Oncogênicas/metabolismo , Transdução de Sinais/fisiologia , Esteroide 16-alfa-Hidroxilase/genética , Transativadores/fisiologia , Animais , Northern Blotting , Família 2 do Citocromo P450 , Perfilação da Expressão Gênica , Hipofisectomia , Janus Quinase 2 , Fígado/efeitos dos fármacos , Fígado/fisiologia , Masculino , Proteínas do Leite , Proteína Quinase 1 Ativada por Mitógeno/metabolismo , Proteína Quinase 3 Ativada por Mitógeno/metabolismo , Fosforilação , Ratos , Ratos Sprague-Dawley , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Fator de Transcrição STAT5 , Transdução de Sinais/efeitos dos fármacosRESUMO
An LH-receptor binding inhibitor (LHRBI), previously isolated from sheep corpora lutea in a partially purified form was subjected to wheat germ lectin chromatography. The unadsorbed fraction (UM-2R-III A) thus obtained had maximum LHRBI activity. This preparation was utilized to develop polyclonal antibodies. The purified fraction could be radioiodinated, suggesting its peptide nature. Intravenous injections of UM-2R-IIIA at 200 microg protein per dose in two doses per day at 9.30 h and 16.30 h on days 1 and 2 of the menstrual cycle to regularly cycling monkeys resulted in a shortening of the length of the cycle by 2 to 3 days. In addition, serum levels of progesterone increased prior to ovulation and remained high through the cycle of all three treated monkeys. It is possible that LHRBI induced enhanced synthesis and/or secretion of progesterone by the ovarian follicles thus suggesting a role for LHRBI in ovarian function.
Assuntos
Peptídeos/administração & dosagem , Animais , Corpo Lúteo , Feminino , Injeções Intravenosas , Peptídeos e Proteínas de Sinalização Intercelular , Macaca mulatta , Ciclo Menstrual/efeitos dos fármacos , Ovário/fisiologia , Peptídeos/isolamento & purificação , Progesterona/sangue , OvinosRESUMO
Neonatal administration of monosodium glutamate (MSG) can produce latently expressed defects in drug metabolism and growth hormone secretion as well as stunted growth and obesity. Instead of secreting growth hormone in the masculine episodic profile, plasma hormone levels are generally undetectable in affected adult male rats. Moreover, male-specific isoforms of cytochrome P450 (P450; e.g., CYP2C11 and CYP3A2), whose combined levels comprise the bulk of the total hepatic P450 in adult male rats, are similarly undetectable in these animals. Since "signaling elements" in the masculine episodic growth hormone profile are solely responsible for the elevated characteristic male-like expression levels of CYP2C11 and CYP3A2, suppression of the isoforms in the MSG-treated rats appeared to be caused by the simple absence of the hormone from the circulation. However, the reported failures of restored physiologic masculine growth hormone profiles to correct the P450 defects suggested the occurrence of direct MSG-induced liver damage independent of the well known hypothalamic lesions produced by the amino acid. Concurrent administration of dizocilpine maleate (MK-801), a selective and highly potent noncompetitive N-methyl-D-aspartate receptor antagonist of glutamate, completely prevented the adverse effects of neonatal MSG treatment on P450 expression, growth hormone secretion, and growth parameters, indicating that the amino acid-induced defects are solely a result of neuronal (i.e., hypothalamic) damage produced at the time of MSG exposure. The irreversibility of the P450 damage is described as resulting from secondary defects initially induced by the neuronal lesions.