RESUMO
Prolactin (PRL) is a potent liver mitogen and proangiogenic hormone. Here, we used hyperprolactinemic rats and PRL receptor-null mice (PRLR(-/-)) to study the effect of PRL on liver growth and angiogenesis before and after partial hepatectomy (PH). Liver-to-body weight ratio (LBW), hepatocyte and sinusoidal endothelial cell (SEC) proliferation, and hepatic expression of VEGF were measured before and after PH in hyperprolactinemic rats, generated by placing two anterior pituitary glands (AP) under the kidney capsule. Also, LBW and hepatic expression of IL-6, as well as suppressor of cytokine signaling-3 (SOCS-3), were evaluated in wild-type and PRLR(-/-) mice before and after PH. Hyperprolactinemia increased the LBW, the proliferation of hepatocytes and SECs, and VEGF hepatic expression. Also, liver regeneration was increased in AP-grafted rats and was accompanied by elevated hepatocyte and SEC proliferation, and VEGF expression compared with nongrafted controls. Lowering circulating PRL levels with CB-154, an inhibitor of AP PRL secretion, prevented AP-induced stimulation of liver growth. Relative to wild-type animals, PRLR(-/-) mice had smaller livers, and soon after PH, they displayed an approximately twofold increased mortality and elevated and reduced hepatic IL-6 and SOCS-3 expression, respectively. However, liver regeneration was improved in surviving PRLR(-/-) mice. PRL stimulates normal liver growth, promotes survival, and regulates liver regeneration by mechanisms that may include hepatic downregulation of IL-6 and upregulation of SOCS-3, increased hepatocyte proliferation, and angiogenesis. PRL contributes to physiological liver growth and has potential clinical utility for ensuring survival and regulating liver mass in diseases, injuries, or surgery of the liver.
Assuntos
Hiperprolactinemia/sangue , Interleucina-6/metabolismo , Regeneração Hepática , Fígado/irrigação sanguínea , Fígado/metabolismo , Neovascularização Fisiológica , Prolactina/sangue , Proteínas Supressoras da Sinalização de Citocina/metabolismo , Animais , Proliferação de Células , Modelos Animais de Doenças , Células Endoteliais/imunologia , Células Endoteliais/metabolismo , Células Endoteliais/patologia , Feminino , Hepatectomia , Hiperprolactinemia/imunologia , Hiperprolactinemia/patologia , Hiperprolactinemia/fisiopatologia , Fígado/imunologia , Fígado/patologia , Fígado/cirurgia , Masculino , Camundongos , Camundongos da Linhagem 129 , Camundongos Knockout , Adeno-Hipófise/metabolismo , Adeno-Hipófise/transplante , Ratos , Ratos Wistar , Receptores da Prolactina/genética , Receptores da Prolactina/metabolismo , Transdução de Sinais , Proteína 3 Supressora da Sinalização de Citocinas , Fatores de Tempo , Fator A de Crescimento do Endotélio Vascular/metabolismoRESUMO
Acting at the level of the brain, interleukin- (IL-)1 ß is considered to be one of the most potent downregulators of reproduction processes during immune/inflammatory challenge. IL-1 ß suppresses gonadotropin-releasing hormone (GnRH) secretion from the hypothalamus resulting in the inhibition of the luteinizing hormone (LH) release from the anterior pituitary (AP). However, the presence of IL-1 ß receptors in the AP suggests the possible direct action of this cytokine on LH secretion. The study was designed to determine the effect of IL-1 ß on the LH secretion from the AP explants collected from saline and LPS-treated ewes in the follicular phase. It was found that IL-1 ß suppressed (P ≤ 0.01) GnRH-stimulated LH release and LH ß gene expression in AP explants in both groups. However, IL-1 ß action was more potent in the explants collected from LPS-treated animals. Pituitaries from LPS-treated animals were characterized by increased (P ≤ 0.01) IL-1 type I receptor and decreased (P ≤ 0.01) GnRH receptor gene expression level compared to the saline-treated group. IL-1 ß also affected the GnRH-R gene expression in explants collected from LPS-treated animals. Our results show that direct action of IL-1 ß on the pituitary gonadotropes could be one of the reasons of the reproductive processes disorders accompanying an inflammatory state.
Assuntos
Inflamação/imunologia , Inflamação/metabolismo , Interleucina-1beta/metabolismo , Lipopolissacarídeos/imunologia , Hormônio Luteinizante/metabolismo , Adeno-Hipófise/imunologia , Adeno-Hipófise/metabolismo , Animais , Regulação da Expressão Gênica/efeitos dos fármacos , Inflamação/induzido quimicamente , Inflamação/genética , Interleucina-1beta/farmacologia , Hormônio Luteinizante/genética , Adeno-Hipófise/transplante , Receptores Tipo I de Interleucina-1/genética , Receptores Tipo I de Interleucina-1/metabolismo , Receptores LHRH/genética , Receptores LHRH/metabolismoRESUMO
The histological, immunohistochemical and ultrastructural alterations in 81 pituitary allografts from Lewis rats transplanted beneath the renal capsule of Wistar rats were investigated. Intrasellar pituitaries of rats bearing allografts were also examined. Recipient rats were sacrificed at various time points after transplantation. Two days after transplantation, the central portion of the allografts demonstrated ischaemic necrosis. A week later, massive mononuclear cell infiltrates consisting primarily of lymphocytes and to a lesser extent, macrophages, plasma cells and granulocytes became prominent. At about three to four weeks after transplantation, the mononuclear cell infiltrate diminished; the surviving adenohypophysial cells, mainly prolactin (PRL) cells, increased in number and necrosis was replaced by connective tissue. No histological changes were noted in the intrasellar pituitaries of rats bearing allografts. Immunohistochemistry demonstrated that the surviving adenohypophysial cells were mainly PRL-producing cells. Electron microscopy revealed adenohypophysial cell destruction, a spectrum of inflammatory cells and, in late phase, accumulation of fibroblasts and collagen fibres. PRL cells were the prominent cell types; they increased in number. It appears that pituitary allografts are 'foreign' and evoke an immune response, suggesting that they may be used as an experimental animal model for morphological investigation of the development and progression of adenohypophysitis, a rare disease occurring mainly in young women often associated with pregnancy.
Assuntos
Rejeição de Enxerto/imunologia , Rejeição de Enxerto/patologia , Doenças da Hipófise/imunologia , Doenças da Hipófise/patologia , Adeno-Hipófise , Animais , Progressão da Doença , Feminino , Rejeição de Enxerto/metabolismo , Hiperplasia , Rim/cirurgia , Masculino , Necrose , Doenças da Hipófise/metabolismo , Adeno-Hipófise/imunologia , Adeno-Hipófise/patologia , Adeno-Hipófise/transplante , Prolactina/metabolismo , Ratos , Ratos Endogâmicos Lew , Ratos Wistar , Transplante HomólogoRESUMO
Estrogens are known to cause pituitary enlargement and lactotroph proliferation. They also modulate pituitary angiogenesis and induce tumor formation. Pituitary grafts, due to the loss of hypothalamic dopamine, also show lactotroph hyperplasia. We investigated the role of estrogen on rat pituitary autograft vascularization by light and transmission electron microscopy, and assessed prolactin (PRL) blood levels, microvessel density (MVD) and cell proliferation using the BrdU labeling index. All adenohypophysial cell types were identified by immunohistochemistry (streptavidin-biotin-peroxidase complex method). The proangiogenic factors, vascular endothelial growth factor (VEGF), its receptor Flk-1, and hypoxia inducible factor-1alpha (HIF-1alpha) were similarly demonstrated. The prevalence of lactotrophs, as well as more intense staining for VEGF, Flk-1 and HIF-1alpha, was noted in those grafts exposed to estrogen, mainly in the area surrounding the central necrotic core. Immunostaining showed Flk-1 expression increased in endothelial cells of the estrogen-exposed grafts as compared with those unexposed. In contrast to the grafts not exposed to estrogen, in the estrogen-exposed grafts, only fenestrated endothelium could be demonstrated, suggesting that estrogen induces fenestration of newly formed capillaries. There was an increase in blood PRL levels in the estrogen-treated groups as compared with controls. Both MVD and BrdU labeling indices were higher in grafts exposed to estrogen, especially after 4 weeks. Our results suggest that estrogen administration not only enhances the expression of proangiogenic factors in the pituitary grafts but also induces their expression at earlier stages, leading to rapid neoformation of purely fenestrated capillaries.
Assuntos
Estradiol/farmacologia , Processamento de Imagem Assistida por Computador , Adeno-Hipófise/irrigação sanguínea , Adeno-Hipófise/transplante , Animais , Biomarcadores/análise , Proliferação de Células/efeitos dos fármacos , Subunidade alfa do Fator 1 Induzível por Hipóxia/análise , Imuno-Histoquímica , Masculino , Microcirculação , Microscopia Eletrônica de Transmissão , Neovascularização Fisiológica/efeitos dos fármacos , Adeno-Hipófise/ultraestrutura , Prolactina/sangue , Ratos , Ratos Wistar , Transplante Autólogo , Fator A de Crescimento do Endotélio Vascular/análise , Receptor 2 de Fatores de Crescimento do Endotélio Vascular/análiseRESUMO
Chronic prolactin (PRL) exposure can affect several functions of duodenal epithelia, especially those associated with fluid and electrolyte transport. However, little is known regarding its molecular mechanism. To identify PRL-regulated genes, microarray analysis was performed on RNA samples from duodenal epithelial cells of anterior pituitary (AP)-grafted hyperprolactinemic rats. Herein, we identified 321 transcripts upregulated and 241 transcripts downregulated after 4 weeks of AP transplantation. Results from real-time PCR analyses of 15 selected genes were consistent with the microarray results. Gene ontology analysis demonstrated pleiotropic effects of PRL on several cellular processes, including cellular metabolic process, cell communication and cell adhesion. Interestingly, 17 upregulated transcripts and 12 downregulated transcripts are involved in the transport of ions and nutrients, e.g., Ca(2+), Na(+), K(+), Cl(-) and glucose, thus agreeing with the established action of PRL on electrolyte homeostasis. The present results provided fundamental information for further investigations on mechanism of PRL actions in the intestine.
Assuntos
Duodeno/efeitos dos fármacos , Perfilação da Expressão Gênica , Mucosa Intestinal/efeitos dos fármacos , Adeno-Hipófise/transplante , Prolactina/farmacologia , Animais , Análise por Conglomerados , Duodeno/metabolismo , Células Epiteliais/efeitos dos fármacos , Células Epiteliais/metabolismo , Feminino , Regulação da Expressão Gênica/efeitos dos fármacos , Mucosa Intestinal/metabolismo , Análise de Sequência com Séries de Oligonucleotídeos , Adeno-Hipófise/metabolismo , Prolactina/metabolismo , Ratos , Ratos Sprague-Dawley , Receptores da Prolactina/genética , Receptores da Prolactina/metabolismo , Transplante Homólogo/fisiologia , Transplante Homólogo/veterináriaRESUMO
Hyperprolactinemia leads to high bone turnover as a result of enhanced bone formation and resorption. Although its osteopenic effect has long been explained as hyperprolactinemia-induced hypogonadism, identified prolactin (PRL) receptors in osteoblasts suggested a possible direct action of PRL on bone. In the present study, we found that hyperprolactinemia induced by anterior pituitary transplantation (AP), with or without ovariectomy (Ovx), had no detectable effect on bone mineral density and content measured by dual-energy X-ray absorptiometry (DXA). However, histomorphometric studies revealed increases in the osteoblast and osteoclast surfaces in the AP rats, but a decrease in the osteoblast surface in the AP+Ovx rats. The resorptive activity was predominant since bone volume and trabecular number were decreased, and the trabecular separation was increased in both groups. Estrogen supplement (E2) fully reversed the effect of estrogen depletion in the Ovx but not in the AP+Ovx rats. In contrast to the typical Ovx rats, bone formation and resorption became uncoupled in the AP+Ovx rats. Therefore, hyperprolactinemia was likely to have some estrogen-independent and/or direct actions on bone turnover. Osteoblast-expressed PRL receptor transcripts and proteins shown in the present study confirmed our hypothesis. Furthermore, we demonstrated that the osteoblast-like cells, MG-63, directly exposed to PRL exhibited lower expression of alkaline phosphatase and osteocalcin mRNA, and a decrease in alkaline phosphatase activity. The ratios of receptor activator of nuclear factor kappaB ligand (RANKL) and osteoprotegerin (OPG) proteins were increased, indicating an increase in the osteoclastic bone resorption. The present data thus demonstrated that hyperprolactinemia could act directly on bone to stimulate bone turnover, with more influence on bone resorption than formation. PRL enhanced bone resorption in part by increasing RANKL and decreasing OPG expressions by osteoblasts.
Assuntos
Remodelação Óssea/fisiologia , Osteoblastos/fisiologia , Osteoprotegerina/metabolismo , Prolactina/metabolismo , Ligante RANK/metabolismo , Absorciometria de Fóton , Animais , Biomarcadores/metabolismo , Densidade Óssea , Linhagem Celular , Dexametasona/metabolismo , Estrogênios/administração & dosagem , Estrogênios/metabolismo , Feminino , Glucocorticoides/metabolismo , Humanos , Hiperprolactinemia/metabolismo , Tamanho do Órgão , Osteoblastos/citologia , Osteoclastos/citologia , Osteoclastos/fisiologia , Ovariectomia , Adeno-Hipófise/transplante , Ratos , Ratos Sprague-Dawley , Receptores da Prolactina/metabolismo , Útero/citologia , Útero/metabolismo , Vitamina D/metabolismoRESUMO
BACKGROUND: The prostate is a key gland in the sexual physiology of male mammals. Its sensitivity to steroid hormones is widely known, but its response to prolactin is still poorly known. Previous studies have shown a correlation between sexual behaviour, prolactin release and prostate physiology. Thus, here we used the sexual behaviour of male rats as a model for studying this correlation. Hence, we developed experimental paradigms to determine the influence of prolactin on sexual behaviour and prostate organization of male rats. METHODS: In addition to sexual behaviour recordings, we developed the ELISA procedure to quantify the serum level of prolactin, and the hematoxilin-eosin technique for analysis of the histological organization of the prostate. Also, different experimental manipulations were carried out; they included pituitary grafts, and haloperidol and ovine prolactin treatments. Data were analyzed with a One way ANOVA followed by post hoc Dunnet test if required. RESULTS: Data showed that male prolactin has a basal level with two peaks at the light-dark-light transitions. Consecutive ejaculations increased serum prolactin after the first ejaculation, which reached the highest level after the second, and started to decrease after the third ejaculation. These normal levels of prolactin did not induce any change at the prostate tissue. However, treatments for constant elevations of serum prolactin decreased sexual potency and increased the weight of the gland, the alveoli area and the epithelial cell height. Treatments for transient elevation of serum prolactin did not affect the sexual behaviour of males, but triggered these significant effects mainly at the ventral prostate. CONCLUSION: The prostate is a sexual gland that responds to prolactin. Mating-induced prolactin release is required during sexual encounters to activate the epithelial cells in the gland. Here we saw a precise mechanism controlling the release of prolactin during ejaculations that avoid the detrimental effects produced by constant levels. However, we showed that minor elevations of prolactin which do not affect the sexual behaviour of males, produced significant changes at the prostate epithelium that could account for triggering the development of hyperplasia or cancer. Thus, it is suggested that minute elevations of serum prolactin in healthy subjects are at the etiology of prostate abnormal growth.
Assuntos
Prolactina/fisiologia , Próstata/fisiologia , Comportamento Sexual Animal/fisiologia , Animais , Ritmo Circadiano , Ejaculação/fisiologia , Células Epiteliais/fisiologia , Haloperidol/farmacologia , Injeções Subcutâneas , Masculino , Concentração Osmolar , Adeno-Hipófise/transplante , Prolactina/sangue , Prolactina/metabolismo , Prolactina/farmacologia , Próstata/efeitos dos fármacos , Próstata/metabolismo , Próstata/patologia , Ratos , Ratos WistarRESUMO
Pituitary autotransplantation eliminates direct vascular contact between the hypothalamus and the adenohypophysis, and enables us to study the role of the hypothalamus in regulating adenohypophysial endocrine activity. The aim of this study was to investigate vascularization of the pituitary autografts. Three-month-old male Wistar rats were hypophysectomized, and their adenohypophyses were autotransplanted under the renal capsule. The animals were killed 3 weeks after autotransplantation. The grafts were removed and studied by using histology, immunohistochemistry and transmission electron microscopy. In the central portion of the grafts, organizing necrosis was apparent. The peripheral portion of the graft contained all adenohypophysial cell types, with a predominance of lactotrophs. Vascular endothelial growth factor and hypoxia-inducible factor were expressed in the graft mainly in the perinecrotic areas. Several capillaries inside the grafts were lined by continuous unfenestrated epithelium, while others were lined by fenestrated endothelium, suggesting that neovascularization is the result of two processes: ingrowths of capillaries from the renal capsule to the graft, and neoformation of capillaries from pre-existing adenohypophysial vessels. In conclusion, hypoxia seems to be an important factor in the vascularization of pituitary autografts. Mediated via hypoxia-inducible factor, hypoxia stimulates vascular endothelial growth factor secretion, which plays a crucial role in angiogenesis.
Assuntos
Neovascularização Fisiológica , Adeno-Hipófise/irrigação sanguínea , Adeno-Hipófise/transplante , Animais , Biomarcadores/análise , Hipofisectomia , Fator 1 Induzível por Hipóxia/análise , Imuno-Histoquímica/métodos , Masculino , Microscopia Eletrônica de Transmissão , Microtomia , Necrose , Adeno-Hipófise/ultraestrutura , Ratos , Ratos Wistar , Transplante Autólogo , Fator A de Crescimento do Endotélio Vascular/análiseRESUMO
Prolactin, having been shown to stimulate transcellular active and solvent drag-induced calcium transport in the duodenum of female rats, was postulated to improve duodenal calcium transport in estrogen-deficient rats. The aim of the present study was, therefore, to demonstrate the effects of long-term prolactin exposure produced by anterior pituitary (AP) transplantation on the duodenal calcium transport in young (9-week-old) and adult (22-week-old) ovariectomized rats. We found that ovariectomy did not alter the transcellular active duodenal calcium transport in young and adult rats fed normal calcium diet (1.0% w/w Ca) but decreased the solvent drag-induced duodenal calcium transport from 75.50 +/- 10.12 to 55.75 +/- 4.77 nmol.hr(-1).cm(-2) (P < 0.05) only in adult rats. Long-term prolactin exposure stimulated the transcellular active calcium transport in young and adult AP-grafted ovariectomized rats fed with normal calcium diet by more than 2-fold from 7.56 +/- 0.79 to 16.54 +/- 2.05 (P < 0.001) and 9.78 +/- 0.72 to 15.99 +/- 1.75 (P < 0.001) nmol.hr(-1).cm(-2), respectively. However, only the solvent drag-induced duodenal calcium transport in young rats was enhanced by prolactin from 95.51 +/- 10.64 to 163.20 +/- 18.03 nmol.hr(-1).cm(-2) (P < 0.001) whereas that in adult rats still showed a decreased flux from 75.50 +/- 10.12 to 47.77 +/- 5.42 nmol.hr(-1).cm(-2) (P < 0.05). Because oral calcium supplement has been widely used to improve calcium balance in estrogen-deficient animals, the effect of a high-calcium diet (2.0% w/w Ca) was also investigated. The results showed that stimulatory action of long-term prolactin on the transcellular active duodenal calcium transport in both young and adult rats was diminished after being fed a high-calcium diet. The same diet also abolished prolactin-enhanced solvent drag-induced duodenal calcium transport in young and further decreased that in adult AP-grafted ovariectomized rats. We concluded that the solvent drag-induced duodenal calcium transport in adult rats was decreased after ovariectomy. Long-term prolactin exposure stimulated the transcellular active duodenal calcium transport in both young and adult rats whereas enhancing the solvent drag-induced duodenal calcium transport only in young rats. Effects of prolactin were abolished by a high-calcium diet.
Assuntos
Cálcio da Dieta/administração & dosagem , Cálcio/metabolismo , Duodeno/metabolismo , Ovariectomia , Adeno-Hipófise/metabolismo , Prolactina/metabolismo , Animais , Cálcio/farmacologia , Feminino , Transporte de Íons/efeitos dos fármacos , Técnicas de Cultura de Órgãos , Adeno-Hipófise/transplante , Ratos , Ratos Sprague-Dawley , Solventes/farmacologia , Fatores de TempoRESUMO
Estradiol is known to increase lactotropic cell proliferation, but estradiol susceptibility varies among human populations and among various strains of rats. We had reported that folliculostellate (FS) cells regulate estradiol's mitogenic action on lactotropes; therefore, we studied their role in determining the susceptibility to estradiol in a high estradiol-responsive rat strain, Fischer 344 (F344), and in a low-responsive strain, Sprague Dawley (SD). Determination of total S-100-positive FS cells in the pituitary revealed that F344 rats have significantly more FS cells than do SD rats. Estradiol treatment did not change the number of FS cells in both F344 and SD rats. When cotransplanted with F344 pituitaries under the kidney capsule or cocultured with F344-derived lactotropes in vitro, FS cells derived from F344 rats increased estradiol's mitogenic action. They also increased estradiol's mitogenic action on SD-derived lactotropes in primary cultures. However, SD-derived FS cells failed to increase estrogen's action on F344- or SD-derived lactotropes. The levels of basic fibroblast growth factor production and secretion by TGF-beta 3 and estradiol were much higher in F344-derived FS cells than in SD-derived FS cells. However, the lactotropes' growth response to basic fibroblast growth factor was similar in both strains. These data suggest that cell-cell interaction between FS cells and lactotropes regulates estradiol's mitogenic action on lactotropes and also determines lactotrope susceptibility to the steroid.
Assuntos
Estradiol/farmacologia , Mitógenos/farmacologia , Adeno-Hipófise/citologia , Adeno-Hipófise/efeitos dos fármacos , Animais , Contagem de Células , Divisão Celular/efeitos dos fármacos , Células Cultivadas , Técnicas de Cocultura , Feminino , Fator 2 de Crescimento de Fibroblastos/metabolismo , Fator 2 de Crescimento de Fibroblastos/farmacologia , Rim , Adeno-Hipófise/transplante , Ratos , Ratos Endogâmicos F344 , Ratos Sprague-Dawley , Fator de Crescimento Transformador beta/farmacologia , Fator de Crescimento Transformador beta3RESUMO
Uterine adenomyosis is a disease in which hyperplastic endometrial stroma and glands invade the myometrium. We have previously demonstrated that hyperprolactinemia leads to the development of adenomyosis in mice. In the present study, a subtracted cDNA library was made by suppression subtractive hybridization to find specific genes that are abundantly expressed in the adenomyotic but not normal tissue in mice. A cDNA fragment of integrinbeta1 (ibeta1) was found in the library, and the expression of the gene product was increased in the adenomyotic uteri at mRNA and protein levels. Intense ibeta1-immunoreactivity was localized on a group of cells dispersing throughout the endometrial stroma. The number of ibeta1-immunoreactive (ibeta1-ir) cells was significantly greater in the uteri of mice with adenomyosis than normal mice. The majority of the ibeta1-ir cells expressed CD14-ir signal, a marker for monocyte-lineage cells, whereas an increase in the number of CD14-ir cells was also evident in the adenomyotic uteri, especially in the ectopic endometrial tissue. Thus, the adenomyotic stromal tissue contained numerous monocyte-lineage cells with higher expression levels of ibeta1, one of their products. The relationship between the increased number of monocyte-lineage cells and the hyperplastic proliferation of endometrial tissues was discussed with a view to understanding the progressive mechanism of adenomyosis.
Assuntos
Endometriose/patologia , Integrina beta1/metabolismo , Monócitos/patologia , Doenças Uterinas/patologia , Animais , Contagem de Células , Linhagem da Célula , Modelos Animais de Doenças , Endometriose/etiologia , Endometriose/metabolismo , Feminino , Biblioteca Gênica , Imuno-Histoquímica , Integrina beta1/genética , Receptores de Lipopolissacarídeos/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos , Monócitos/metabolismo , Adeno-Hipófise/transplante , RNA Mensageiro/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Transplantes , Doenças Uterinas/etiologia , Doenças Uterinas/metabolismoRESUMO
The possibility of therapeutic application of novel nonsteroidal progesterone receptor modulators CP8816 and CP8863 for preventing the development of uterine adenomyosis was investigated in mice. First priming effects of CP8816 on 17beta-estradiol (E2)-induced cell division in uterine tissues were examined. As a result, pretreatment with CP8816 or progesterone significantly suppressed the elevation of the mitotic activity in the luminal epithelial cells of mice treated with E2 later. Priming with CP8816 had little effect on the stromal cells, but progesterone priming caused an increase of stromal mitotic activity in mice treated with E2 later. To evaluate the inhibitory effect of these compounds on the development of adenomyosis induced experimentally by pituitary grafting, 7-week-old female mice were isografted with a single anterior pituitary in the uterus and divided into four groups. Two groups of mice were given daily subcutaneous injections of 1 mg of CP8816 or the vehicle alone for 6 weeks from the day after the grafting. Remaining two groups of mice were given oral administration of 1 mg of CP8863 or the vehicle only for 5 weeks starting one week after the grafting. The incidence of adenomyosis was significantly lower in the groups of mice treated with CP8816 and CP8863 than in the respective control groups. The mechanism by which CP compounds inhibited the development of adenomyosis might be related to their priming effects, i.e., their inhibitory effect on epithelial cell division and lack of effect on stromal cell division after subsequent exposure to E2.
Assuntos
Endometriose/prevenção & controle , Indóis/farmacologia , Receptores de Progesterona/metabolismo , Sesquiterpenos , Útero/efeitos dos fármacos , Animais , Endometriose/patologia , Estradiol/farmacologia , Feminino , Furanos/química , Humanos , Ligantes , Masculino , Camundongos , Naftóis/química , Ovariectomia , Adeno-Hipófise/transplante , Progesterona/sangue , Útero/patologiaRESUMO
Folliculo-stellate cells (FS-cells) in the anterior pituitary gland are star-shaped cells and form tiny follicles. FS-cells are positive for S-100 protein and produce many cytokines or growth factors, such as interleukin-6 (IL-6), leukemia inhibitory factor (LIF), basic fibroblastic growth factor (bFGF) and vascular endothelial cell growth factor (VEGF). Therefore, it is generally accepted that FS-cells regulate endocrine cells through these growth factors. FS-cells also exhibit a phagocytotic activity and are known to work as scavenger cells. In addition to these functions, FS-cells are considered to have some unknown functions. In order to reveal the biological significance of FS-cells in the anterior pituitary gland, we performed a morphological study and obtained some new findings. First, we were interested in the colloid formation in the senescent porcine pituitary gland. We analyzed the colloids and found that clusterin is a major protein in them. We also found that the accumulation of clusterin in the colloids is related to the phagocytotic activity of FS-cells. In our next study, we found that FS-cells have the potential to differentiate into striated muscle cells. From FS-cells show multi-potent cell character and other cytological evidence, we propose that FS-cells are candidate of organ-specific stem cells in the anterior pituitary gland.
Assuntos
Adeno-Hipófise/citologia , Células-Tronco/fisiologia , Animais , Clusterina , Glicoproteínas/fisiologia , Chaperonas Moleculares/fisiologia , Músculo Esquelético/citologia , Músculo Esquelético/ultraestrutura , Especificidade de Órgãos , Adeno-Hipófise/fisiologia , Adeno-Hipófise/transplante , Adeno-Hipófise/ultraestruturaRESUMO
BACKGROUND: To evaluate the effects of hyperprolactinemia on aldosterone secretion and its mechanisms of action in ovariectomized (OVX) rats. METHODS: Hyperprolactinemia was induced by the transplantation of rat anterior pituitary (AP) glands under the kidney capsule for 6 weeks in female rats. Control rats underwent cerebral cortex (CX) transplantation. Four weeks after transplantation, the rats were OVX 2 weeks before decapitation. After decapitation, the trunk blood was collected, and the adrenal glands of CX- and AP-grafted rats were prepared as zona glomerulosa (ZG) cells for in vitro study. RESULTS: Plasma prolactin and aldosterone in the rats were increased by AP gland transplantation. In the in vitro study, the basal aldosterone secretion by the adrenal ZG cells was higher in AP-grafted rats than in CX-grafted rats. The AP-grafted group showed increased responsiveness to angiotensin II (10(-8) M), KCl (8 x 10(-3) M), or 8-bromo-adenosine 3',5'-cyclic monophosphate (8-br-cAMP; 10(-4) M, a membrane-permeable analogue of cAMP) with regard to aldosterone secretion as compared with the CX-grafted group. N-(2-[p-Bromocinnamylamine]ethyl)-5-isoquinolinesulfonamide (H89; 10(-6), 10(-5) M, a protein kinase A inhibitor) or tetrandrine (10(-5) M, a blocker for both L-type and T-type Ca2+ channels) induced a greater suppression of aldosterone secretion in the AP-grafted group than in the CX-grafted group. No significant differences between the CX- and AP-grafted groups were observed, however, with regard to the adrenocorticotropichormone (10(-9) M)-, forskolin (10(-5) M, an adenylyl cyclase activator)-, or nifedipine (10(-5) M, an L-type Ca2+ channel blocker)-induced responsiveness of aldosterone secretion. In addition, there was no difference in the expression of desmolase (i.e., cytochrome P450 side-chain cleavage enzyme) in ZG cells between AP- and CX-grafted rats. The conversions of 25-OH-cholesterol into pregnenolone in the presence of trilostane (an inhibitor of 3beta-hydroxysteroid dehydrogenase) and corticosterone into aldosterone, as well as the expression of the steroidogenic acute regulatory protein in ZG cells, were greater in AP-grafted rats than in CX-grafted rats. CONCLUSIONS: These results suggest that hyperprolactinemia increases basal, angiotensin II- and KCl-stimulated aldosterone secretion by ZG cells in OVX rats through activation of T-type Ca2+ channels, the post-cAMP and protein kinase A pathway, cytochrome P450 side-chain cleavage enzyme, and aldosterone synthase, as well as by causing increased expression of steroidogenic acute regulatory protein in ZG cells.
Assuntos
Aldosterona/sangue , Benzilisoquinolinas , Hiperprolactinemia/metabolismo , Sulfonamidas , 8-Bromo Monofosfato de Adenosina Cíclica/farmacologia , Adenilil Ciclases/metabolismo , Alcaloides/farmacologia , Angiotensina II/farmacologia , Animais , Canais de Cálcio Tipo T/metabolismo , Células Cultivadas , Enzima de Clivagem da Cadeia Lateral do Colesterol/metabolismo , Proteínas Quinases Dependentes de AMP Cíclico/metabolismo , Citocromo P-450 CYP11B2/metabolismo , Feminino , Hiperprolactinemia/etiologia , Isoquinolinas/farmacologia , Ovariectomia , Fosfoproteínas/metabolismo , Adeno-Hipófise/transplante , Cloreto de Potássio/farmacologia , Prolactina/sangue , Ratos , Ratos Sprague-Dawley , Zona Glomerulosa/citologia , Zona Glomerulosa/efeitos dos fármacos , Zona Glomerulosa/metabolismoRESUMO
The histology and porphyrin concentrations of Harderian glands and plasma prolactin levels were examined in B6C3F1 mice castrated or isografted with pituitaries in combination with the administration of bromocriptine (2-bromo-á-ergocryptine), a potent suppressor of prolactin. Four pituitaries were transplanted underneath the bilateral kidney capsules of each mouse. Furthermore, we investigated Harderian porphyrins and plasma testosterone levels in mice that were castrated or treated with neuroleptic butyrophenone (timiperone), a potent accelerator of prolactin, and treated concurrently with bromocriptine. Light microscopically, pituitary grafts increased the concretion of porphyrin pigments within the Harderian gland lumina. Pituitary grafts or castration increased both Harderian porphyrin concentrations and plasma prolactin levels compared with the intact control mice. In pituitary-grafted or castrated mice, bromocriptine distinctly prevented the rise in both porphyrins and prolactin levels. Administration of butyrophenone did not result in any marked change in testosterone levels, although Harderian gland porphyrins were significantly increased. The present results indicate that in mice prolactin stimulates the porphyrin production of the Harderian gland and has an important role in the regulation of Harderian gland porphyrins.
Assuntos
Glândula de Harder/metabolismo , Hiperprolactinemia/metabolismo , Adeno-Hipófise/metabolismo , Animais , Peso Corporal , Bromocriptina/farmacologia , Butirofenonas/farmacologia , Modelos Animais de Doenças , Feminino , Glândula de Harder/efeitos dos fármacos , Antagonistas de Hormônios/farmacologia , Hiperprolactinemia/induzido quimicamente , Hiperprolactinemia/patologia , Masculino , Camundongos , Camundongos Endogâmicos , Orquiectomia , Tamanho do Órgão/efeitos dos fármacos , Transplante de Órgãos , Adeno-Hipófise/transplante , Porfirinas/metabolismo , Prolactina/sangue , Testículo/efeitos dos fármacos , Testículo/patologia , Testosterona/sangueRESUMO
The existence of adenosine A1 receptors and adenosine transporters in the central nervous system has been well demonstrated, although their possible modulation by hormones and/or exogenous drugs is poorly understood. To further analyze these modulatory mechanisms, the effects of prolactin and cyclosporine (CyA) on adenosine A1 receptors and transporters were analyzed in the central nervous system. For this purpose the number and affinity of adenosine A1 receptors were measured using the specific antagonist 1,3-dipropyl-8-cyclopentylxanthine (DPCPX) and the transporters with the high affinity ligand nitrobenzylthioinosine (NBTI). This procedure was carried out in hyperprolactinemic and control male rats treated with CyA or its vehicle for 8 days. As expected, pituitary grafting increased plasma prolactin levels (p<0.01). CyA treatment reduced but did not normalize (p<0.05) this parameter in hyperprolactinemic rats and did not modify circulating prolactin in control animals. Both hyperprolactinemia and CyA treatment reduced the number of adenosine transporters by 70% and by 40% the number of A1 receptors. The Kd for transporters was also reduced in all experimental groups. Hyperprolactinemia increased the affinity of A1 receptors (p<0.01) and CyA treatment did not further modify this parameter. These data demonstrated that prolactin and CyA influence adenosine transporters and A1 receptors at the central nervous system and suggest the existence of an interaction between prolactin and CyA may be operating to modulate these processes.
Assuntos
Adenosina/metabolismo , Encéfalo/metabolismo , Proteínas de Transporte/metabolismo , Ciclosporina/farmacologia , Proteínas de Membrana/metabolismo , Prolactina/fisiologia , Receptores Purinérgicos P1/metabolismo , Tioinosina/análogos & derivados , Animais , Encéfalo/efeitos dos fármacos , Feminino , Hiperprolactinemia/induzido quimicamente , Masculino , Proteínas de Transporte de Nucleosídeos , Adeno-Hipófise/transplante , Prolactina/sangue , Antagonistas de Receptores Purinérgicos P1 , Ensaio Radioligante , Ratos , Ratos Wistar , Tioinosina/metabolismo , Tioinosina/farmacologia , Xantinas/metabolismo , Xantinas/farmacologiaRESUMO
This study was designed to investigate the effect of hyperprolactinemia, with high or low estrogen levels, on the response to imipramine in the forced swimming test. Three groups of female rats were studied: (1) ovariectomized controls, with low serum prolactin (PRL) and estrogen levels, (2) ovariectomized, estrogen-treated rats, with high PRL and high estrogen levels, and (3) pituitary-grafted rats, with high PRL and low estrogen levels. The hyperprolactinemic groups did not show significant behavioral changes in the forced swimming test preceded by saline injection. Imipramine decreased the immobility time by 37.5% in ovariectomized controls but not in the pituitary-grafted group, and there was an increment of 48.4% in immobility time following imipramine administration in the estrogen-treated group (p<0.05). This paradoxical response to imipramine was significantly correlated with serum PRL (r = 0.59, p<0.01) but not with estradiol levels. These findings suggest that, at least in female rats submitted to the forced swimming model, PRL may induce reversed behavioral effects in response to imipramine, independently of circulating estrogen levels.
Assuntos
Hiperprolactinemia/fisiopatologia , Imipramina/farmacologia , Natação/fisiologia , Animais , Comportamento Animal/efeitos dos fármacos , Comportamento Animal/fisiologia , Estradiol , Feminino , Hiperprolactinemia/induzido quimicamente , Locomoção/efeitos dos fármacos , Locomoção/fisiologia , Ovariectomia , Adeno-Hipófise/metabolismo , Adeno-Hipófise/transplante , Prolactina/sangue , Ratos , Ratos Wistar , Tempo de Reação/efeitos dos fármacos , Tempo de Reação/fisiologiaRESUMO
The role of prolactin (PRL) in calcitonin (CT) release by the thyroid C cell in male rats was studied. Anterior pituitary (AP)-grafted male rats were characterized by hyperprolactinemia. Brain cortex (CX)-grafted male rats were used as control animals. AP- and CX-grafted rats were infused intravenously with CaCl2 and bled from the jugular catheter at 0, 30, 60, and 120 minutes following the CaCl2 challenge. Rat thyroid gland was incubated with or without 3-isobutyl-1-methylxanthine (IBMX) at 37 degrees C for 30 minutes. Thyroid C cells were incubated in culture medium at 37 degrees C for 60 minutes. Cyclic adenosine 3',5'-monophosphate (cAMP) in rat thyroid tissues following incubation with IBMX was extracted by 65% ethanol. AP-grafted rats had higher plasma levels of PRL and CT compared with CX-grafted rats. Both the release of CT and accumulation of cAMP in thyroid glands were higher in AP-grafted versus CX-grafted rats. Direct administration of ovine PRL (oPRL) on the thyroid glands did not increase CT secretion in vitro. Thyroid C cells of AP-grafted rats secreted more CT compared with CX-grafted rat cells. These results suggest that hyperprolactinemia increases the release of CT by thyroid C cells in rats through a cAMP-dependent pathway caused by an indirect effect of PRL.
Assuntos
Calcitonina/sangue , Hiperprolactinemia/sangue , Animais , Células Cultivadas , AMP Cíclico/metabolismo , Masculino , Adeno-Hipófise/transplante , Ratos , Ratos Sprague-Dawley , Glândula Tireoide/citologia , Glândula Tireoide/metabolismoRESUMO
The role of prolactin (PRL) in the male is not fully defined. The aim of this study was to investigate the function and mechanism of PRL on the production of corticosterone by zona fasciculata-reticularis (ZFR) cells in vitro. The ZFR cells were obtained from male rats under normal, hyperprolactinemic, or hypoprolactinemic situation. PRL stimulated the corticosterone release in a dose-dependent pattern in the ZFR cells from normal male rats. The cellular adenosine 3'-5'-cyclic monophosphate (cAMP) concentration positively correlated with PRL concentration in the presence of forskolin or 3-isobutyl-1-methylxanthine (IBMX). PRL enhanced the stimulatory effects of cAMP mimetic reagents, i.e., forskolin, 8-bromo-adenosine 3',5'-cyclic monophosphate (8-Br-cAMP), and IBMX on the release of corticosterone. The adenylate cyclase inhibitor (SQ22536) inhibited the corticosterone release in spite of presence of PRL. Nifedipine (L-type calcium channel blocker) did not inhibit corticosterone release. The hyperprolactinemic condition was actualized by transplantation of donor rat anterior pituitary glands (APs) under kidney capsule. By comparison with the cerebral cortex (CX)-grafted group, AP-graft resulted in an increased release of corticosterone, 3beta-hydroxysteriod dehydrogenase (HSD) activity and cAMP production by ZFR cells. Acute hypoprolactinemic status was induced by bromocriptine for 2 days. The results showed the productions of corticosterone were lower in hypoprolactinemic group than in control group, which were persistent along with different ACTH concentrations. These results suggest that PRL increase the release of corticosterone by ZFR cells via cAMP cascades and 3beta-HSD activity.
Assuntos
Corticosterona/metabolismo , Prolactina/farmacologia , Zona Fasciculada/metabolismo , Zona Reticular/metabolismo , 1-Metil-3-Isobutilxantina/farmacologia , 8-Bromo Monofosfato de Adenosina Cíclica/farmacologia , Adenina/análogos & derivados , Adenina/farmacologia , Animais , Bromocriptina/administração & dosagem , Células Cultivadas , Córtex Cerebral/transplante , Colforsina/farmacologia , Corticosterona/sangue , AMP Cíclico/biossíntese , Desoxicorticosterona/farmacologia , Hiperprolactinemia/metabolismo , Injeções Subcutâneas , Masculino , Nifedipino/farmacologia , Adeno-Hipófise/transplante , Pregnenolona/farmacologia , Progesterona/farmacologia , Prolactina/sangue , Ratos , Ratos Sprague-Dawley , Ovinos , Zona Fasciculada/citologia , Zona Fasciculada/efeitos dos fármacos , Zona Reticular/citologia , Zona Reticular/efeitos dos fármacosRESUMO
The mechanism that causes hypercalcitonemia in female rats and is associated with aging was investigated. Young (3 mo), adult (8 mo), middle-aged (12 mo), and old (21 mo) rats were infused with CaCl2 and were bled from a jugular catheter after a CaCl2 challenge. To mimic some of the hormonal changes caused by aging, the anterior pituitary (AP)-grafted ovariectomized rats with hyperprolactinemic syndrome were used to mimic the physiological status of aging. The rat thyroid gland was incubated with or without ovine prolactin (oPRL; 40 or 80 ng/ml) at 37 degreesC for 30 min. Old rats possessed the lowest levels of plasma estradiol and progesterone yet had the highest levels of plasma prolactin and calcitonin (CT) compared with young, adult, and middle-aged rats. The basal release of thyroid CT in vitro in thyroid glands gradually increased with age. Compared with cortex (CX)-grafted rats, the AP-grafted rats possessed higher levels of plasma PRL, basal and CaCl2-induced levels of plasma CT, and the release of thyroid CT in thyroid glands. After stimulation with oPRL, the in vitro release of thyroid CT increased in both CX- and AP-grafted rats. These results suggest that the hypersecretion of CT in old rats is due at least in part to hyperprolactinemia.