RESUMO
The genetically obese (ob/ob) mouse is a useful model for the study of the diabetogenic action of growth hormone (GH), because treatment of these animals with GH results in decreased responsiveness of their adipose tissue to insulin in vitro. Studies of the mechanisms involved in GH-induced insulin resistance using isolated adipocytes of ob/ob mice have not been possible, however, because of their extreme fragility and the lack of an adequate system for the maintenance of these cells. This study describes a new method for the isolation of ob/ob mouse adipocytes. The isolated cells are stable, viable and metabolically responsive to insulin. In addition, these adipocytes have been maintained in primary culture, in serum-free medium, for up to 3 days. During culture, the cells exhibit large increases in 125I-hGH binding (10-20-fold) and porcine 125I-insulin binding (5-10-fold). The induction of insulin resistance by GH has also been demonstrated in these freshly isolated ob/ob mouse adipocytes. The studies to date indicate that the ob/ob mouse adipocyte system should provide a useful model for detailed studies of the cellular and molecular mechanisms of GH induced insulin resistance.
Assuntos
Tecido Adiposo/efeitos dos fármacos , Gonadotropinas/farmacologia , Insulina/farmacologia , Tecido Adiposo/citologia , Tecido Adiposo/metabolismo , Animais , Dióxido de Carbono/metabolismo , Células Cultivadas , Resistência a Medicamentos , Glucose/metabolismo , Camundongos , Camundongos ObesosRESUMO
Luteinizing hormone receptor (LHR) is a G-protein-coupled receptor that exerts its effects mainly through increased cAMP synthesis. Our previous studies have shown that a ovarian cytosolic protein, designated as LHR mRNA binding protein (LRBP) is an important regulator of the steady state levels of LHR expression. To test whether LHR mRNA expression is modulated by cAMP through LRBP activity, we used rolipram, a type IV phosphodiesterase inhibitor that is known to promote intracellular cAMP accumulation. On day 4 of pseudopregnancy, rats were treated with rolipram (1.25 mg/injection) to raise intracellular levels of cAMP. In order to maintain higher cAMP levels, up to four injections of rolipram were given, with the last injection 4 h before collecting the ovaries. Measurement of cAMP levels showed an increase (p< or =0.05) at 8, 12, and 24 h after rolipram injections at total dosages of 2.5, 3.75 and 5.0 mg/rat, respectively. Northern blot analysis of LHR mRNA showed that rolipram treatment also markedly reduced ovarian LHR mRNA levels by up to 75%. LHR mRNA binding activity of LRBP, assayed by RNA electrophoretic mobility shift analysis, using S-100 fractions from control or rolipram-treated ovaries showed increased LHR mRNA binding activity in the S-100 fractions from rolipram treated groups. These data indicate that chronic elevation of ovarian cAMP leads to a decreased expression of LHR mRNA with a concomitant increase in LHR mRNA binding activity of LRBP.
Assuntos
Regulação para Baixo , Ovário/metabolismo , Proteínas de Ligação a RNA/metabolismo , Receptores do LH/genética , Animais , AMP Cíclico/metabolismo , Feminino , Ovário/efeitos dos fármacos , Inibidores de Fosfodiesterase/farmacologia , Pseudogravidez/metabolismo , RNA Mensageiro/análise , RNA Mensageiro/metabolismo , Ratos , Rolipram/farmacologiaRESUMO
Analogs of thiazolidinedione improve the responsiveness of insulin-resistant animals to insulin. One such analog, pioglitazone (5-(4-[2-(5-ethyl-2-pyridinyl)ethoxy]benzyl)thiazolidine-2,4-dione hydrochloride), when fed to insulin-resistant animals such as the obese (ob/ob) mouse, reduces blood glucose and lipids and also lowers the plasma insulin level. Because GH can produce insulin resistance in humans and animals such as the ob/ob mouse, the present study was conducted to determine whether feeding pioglitazone can 1) inhibit the ability of GH to induce enhanced insulin resistance in obese mice, 2) ameliorate or reverse GH-induced insulin resistance once it has been induced in ob/ob mice, and 3) alter the ability of GH to promote growth in hypophysectomized rats. Female ob/ob mice were fed a control diet or a diet containing pioglitazone (20 mg/kg animal.day) for 4 days. During the last 3 days of the feeding period, the mice also received a daily sc injection of either saline or 200 micrograms S-carboxymethylated human GH (RCM-hGH), which is a GH derivative having mainly diabetogenic activity. In control-fed mice, RCM-hGH increased blood glucose and plasma insulin levels, which is an expected response to GH-induced insulin resistance. By contrast, the ability of RCM-hGH to increase blood glucose and plasma insulin levels was totally blocked in pioglitazone-fed mice. To determine whether pioglitazone can ameliorate GH-induced insulin resistance once it has been established, ob/ob mice were treated sc with either saline or 200 micrograms RCM-hGH for 3 days. Half of the saline-treated and half of the hormone-treated mice were then fed pioglitazone, whereas the remaining animals were continued on the control diet. After 48 h on the diets, the blood glucose and plasma insulin levels of the RCM-hGH treated mice fed the control diet remained elevated with respect to those in the saline-treated controls. On the other hand, the blood glucose and plasma insulin levels of the RCM-hGH treated mice fed pioglitazone were markedly reduced compared to those of the RCM-hGH-treated control-fed animals. Thus, these results suggest that pioglitazone can ameliorate GH-induced insulin resistance.(ABSTRACT TRUNCATED AT 400 WORDS)
Assuntos
Glicemia/efeitos dos fármacos , Comportamento Alimentar/fisiologia , Hormônio do Crescimento/análogos & derivados , Hipoglicemiantes/farmacologia , Tiazóis/farmacologia , Tiazolidinedionas , Animais , Glicemia/metabolismo , Diabetes Mellitus Experimental/prevenção & controle , Comportamento Alimentar/efeitos dos fármacos , Feminino , Hormônio do Crescimento/farmacologia , Humanos , Hipofisectomia , Resistência à Insulina , Camundongos , Camundongos Obesos , PioglitazonaRESUMO
Recent work with various point and deletion mutants of human GH (hGH) has suggested that the proximal N-terminal end of the hormone molecule is important for its growth promoting action. This study was conducted to examine the growth promoting, diabetogenic, and insulin-like activities of two N-terminal mutants of hGH, the deletion mutant Des-7 hGH (met8, ala11), and a chimeric mutant of bovine GH (bGH) and hGH containing the N-terminal 13 amino acids of bGH (met, ala 1-13/14-191, asp11). The CD spectra of these mutants are similar to that of wild-type hGH and they retain lactogenic activity on Nb2 lymphoma cells, whereas their ability to bind to somatogenic receptors on IM-9 lymphocytes and bovine liver membranes is markedly reduced. In this study, growth promoting activity of the mutants was assessed using the 9-day weight gain test in hypophysectomized rats. Des-7 hGH had a potency of 0.03 IU/mg protein in this assay, whereas the potency of the bGH/hGH chimera was 0.71 IU/mg. Diabetogenic activity was tested in the ob/ob mouse, using the elevation of fasting blood glucose and the worsening of glucose tolerance after a 3-day course of treatment as end-points. Both Des-7 hGH and the bGH/hGH chimera had reduced diabetogenic activity compared to that of biosynthetic wild-type hGH, consistent with their reduced growth activity. Insulin-like activity was assessed by testing the in vitro ability of the mutants to stimulate [14C] glucose oxidation by epididymal adipose tissue of hypophysectomized rats. Des-7 hGH had about 1% the activity of wild-type hGH, whereas the chimera was about 20% as active. When Des-7 hGH was added to the incubation medium along with wild-type hGH in ratios of 5, 12.5, or 25:1 (Des-7 hGH:hGH), the insulin-like action of hGH was significantly inhibited, indicating that the mutant is a modest antagonist of the insulin-like action of hGH. When the ability of Des-7 hGH to compete with [125I] hGH for binding to isolated rat adipocytes was tested, the mutant was about 10% as effective as wild-type hGH. Thus, Des-7 hGH appears to be more effective in binding to adipocyte GH receptors than in triggering an insulin-like response, perhaps accounting for its modest antagonistic activity. The results of this study suggest that the proximal N-terminal end of the hGH molecule is involved in the expression of the growth promoting, diabetogenic and insulin-like activities of GH.
Assuntos
Glicemia/metabolismo , Hormônio do Crescimento/química , Insulina/fisiologia , Aumento de Peso/efeitos dos fármacos , Tecido Adiposo/efeitos dos fármacos , Tecido Adiposo/metabolismo , Tecido Adiposo/ultraestrutura , Animais , Feminino , Hormônio do Crescimento/genética , Hormônio do Crescimento/fisiologia , Masculino , Camundongos , Camundongos Mutantes , Camundongos Obesos , Mutação/genética , Conformação Proteica , Ratos , Receptores da Somatotropina/efeitos dos fármacos , Receptores da Somatotropina/fisiologiaRESUMO
Physiologically, the action of insulin on carbohydrate and lipid metabolism is opposed by several hormones, including glucocorticoids, glucagon, catecholamines, and pituitary GH. Perhaps least is known about the mechanism(s) involved in the antiinsulin action of GH. Since the generation of diacylglycerol (DAG) appears to be an early event in the insulin-signaling cascade, it was of interest to determine whether GH would interfere with this effect of insulin. Experiments were conducted to determine whether insulin would stimulate the generation of DAG in adipocytes of the obese (ob/ob) mouse, and whether this response could be blocked by the diabetogenic GH derivative S-carboxymethylated human GH (RCM-hGH). Isolated adipocytes of the ob/ob mouse were used for these studies, because unlike normal rodents, the ob/ob mouse responds predictably to the antiinsulin action of GH. Insulin produced a rapid biphasic increase in the amount of DAG in a crude membrane fraction of the adipocytes. The first peak in DAG mass occurred within 5 min of exposure of the cells to insulin, and the second peak occurred after 30 min. The first peak in DAG mass did not occur in adipocytes that had been incubated with pertussis toxin before exposure to insulin. Also, adipocytes isolated from ob/ob mice that had been treated with RCM-hGH failed to respond to insulin with an increase in DAG mass. RCM-hGH blocked both the first and second insulin-induced peaks in DAG mass within 6 h of its administration. This is the time at which ob/ob mouse adipocytes exhibit increased insulin resistance in response to RCM-hGH. Neither exposure to insulin nor treatment with RCM-hGH had any appreciable effect on the fatty acid composition of the DAG present in the adipocyte membranes. These findings are compatible with the idea that GH produces some defect in the insulin-signaling cascade that is proximal to the events that result in the generation of DAG in the adipocyte.
Assuntos
Tecido Adiposo/metabolismo , Diglicerídeos/biossíntese , Hormônio do Crescimento/farmacologia , Antagonistas da Insulina/farmacologia , Insulina/farmacologia , Tecido Adiposo/ultraestrutura , Animais , Membrana Celular/metabolismo , Ácidos Graxos/metabolismo , Feminino , Hormônio do Crescimento/análogos & derivados , Camundongos , Camundongos Obesos , Toxina Pertussis , Fatores de Virulência de Bordetella/farmacologiaRESUMO
Puberty in the female lamb is accompanied by an increased frequency of LH pulses, and during normal development this is preceded by a decline in GH. Conversely, in the growth-retarded lamb, when LH levels are depressed by low nutrition, GH secretion is elevated. Based upon this inverse relationship, we tested the hypothesis that GH may act as a metabolic signal from the brain to inhibit the secretion of LH, and that the decline in GH times puberty. Our approach was to extend high circulating GH levels far beyond the early postnatal period, in a physiological pattern and level, in an attempt to block the pubertal LH rise. To evaluate the pattern of LH as a continuous variable under conditions of constant estradiol negative feedback, the gonadotropin was measured in blood samples collected by jugular venipuncture twice weekly; the lambs were ovariectomized and treated chronically with estradiol (Silastic capsule) beginning at 3 weeks of age. Nine lambs served as untreated controls, and 7 were infused iv with pituitary-derived bovine GH (bGH) between 5 and 28 weeks of age. A programmable backpack infusion pump delivered bGH as hourly pulses, with a total dose of 18 micrograms/kg.24 h, to maintain a physiological pattern and level of GH. At various ages, blood samples were collected at 12-min intervals for 6 h to monitor patterns and levels of peripheral LH and GH. Circulating GH in untreated and treated lambs averaged 7.7 +/- 1.5 ng/ml over a 6-h period at 4 weeks of age and declined to 1.1 +/- 0.2 ng/ml by 19 weeks in the untreated lambs; in contrast, bGH-infused lambs averaged 10.4 +/- 0.9 ng/ml at 19 weeks. Although body weights did not differ, back fat depth and quantity of perirenal fat were reduced in bGH-treated females compared to that in controls. Moreover, insulin-like growth factor-I levels were higher in bGH-treated compared with control lambs, and the bGH-treated lambs exhibited glucose intolerance, thus confirming that infused bGH was biologically active. Neuroendocrine sexual maturity, however, was not different in bGH-treated and control lambs, and it occurred at 21-22 weeks of age. The results do not support our hypothesis that decreasing GH secretion is a requirement for puberty in the sheep. Moreover, unlike in children with delayed puberty, exogenous bGH did not advance normal puberty in the lamb.(ABSTRACT TRUNCATED AT 400 WORDS)
Assuntos
Hormônio do Crescimento/administração & dosagem , Reprodução/fisiologia , Animais , Constituição Corporal , Bovinos , Ingestão de Alimentos/efeitos dos fármacos , Feminino , Glucose/farmacocinética , Hormônio do Crescimento/sangue , Hormônio do Crescimento/farmacologia , Fator de Crescimento Insulin-Like I/análise , Hormônio Luteinizante/metabolismo , Sistemas Neurossecretores/crescimento & desenvolvimento , Ovinos/crescimento & desenvolvimento , Ovinos/metabolismo , Ovinos/fisiologia , Fatores de TempoRESUMO
The rabbit corpus luteum possesses LH receptors that are coupled to adenylyl cyclase, but paradoxically it does not require LH as a luteotrophic factor for the maintenance of progesterone secretion. This suggests that rabbit luteal cells may not respond physiologically to LH. Therefore, the present study was undertaken to investigate the responsiveness of the rabbit corpus luteum of pseudopregnancy to human chorionic gonadotrophin (hCG) which acts on the same receptor as LH. Pseudopregnancy was induced by injection of 40 IU pregnant mare serum gonadotrophin followed 50 h later by an injection of 40 IU hCG (day 0). On days 7 and 11 of pseudopregnancy, corpora lutea were obtained and incubated for 2 or 5 h in the presence of either 0.1 or 1 microgram/ml hCG or 1 mM monobutyryl cyclic AMP (bcAMP). Neither hCG nor bcAMP stimulated progesterone production by the isolated corpus luteum, despite a sustained high rate of progesterone production by the tissue throughout the incubation period. By contrast, Graafian follicles removed from the same ovaries and incubated under the same conditions responded both to hCG and bcAMP with large increases in progesterone production. To determine whether the cyclic AMP content of the corpus luteum was altered by in vitro exposure to hCG, day 7 and day 11 corpora lutea were incubated for 5 or 15 min with various concentrations of hCG, and cyclic AMP in the tissue was then measured. Even at the highest concentration of hCG tested (10 micrograms/ml), the cyclic AMP content of the corpus luteum was unaltered.(ABSTRACT TRUNCATED AT 250 WORDS)
Assuntos
Manutenção do Corpo Lúteo/fisiologia , Corpo Lúteo/fisiologia , Hormônio Luteinizante/fisiologia , Adenilil Ciclases/metabolismo , Animais , Gonadotropina Coriônica/farmacologia , Corpo Lúteo/efeitos dos fármacos , Corpo Lúteo/metabolismo , Técnicas de Cultura , AMP Cíclico/metabolismo , Feminino , Folículo Ovariano/efeitos dos fármacos , Gravidez , Progesterona/biossíntese , Coelhos , Receptores do LH/metabolismoRESUMO
OBJECTIVE: Prolactin is capable of both trophic and lytic actions in rat corpora lutea. In corpora lutea responding to a trophic prolactin signal, the long form of the prolactin receptor is the dominant form and is upregulated by prolactin. We investigated whether mRNA for the short form of the prolactin receptor was dominant in corpora lutea responding to a lytic prolactin signal, and whether the relative concentrations of the mRNAs for both forms of the prolactin receptor were changed during this response. DESIGN AND METHODS: Immature rats were ovulated by injection of 5 IU equine chorionic gonadotrophin and 5 IU human chorionic gonadotrophin, and were hypophysectomized shortly after ovulation. Nine days after hypophysectomy, rats were injected with prolactin (500 microg/day) or vehicle for 24 (n=6, n=6) or 72 h (n=13, n=5). Total RNA was isolated from corpora lutea and mRNA for both types of prolactin receptor were analyzed by semiquantitative RT-PCR using the ribosomal protein S16 as the internal control. RESULTS: The intensities of the long- and short-form prolactin receptor signals were normalized to the S16 internal control and expressed as relative densitometric units. The normalized values at 24h for prolactin-treated vs vehicle-treated rats were 0.23 +/- 0.05 vs 0.49 +/- 0.15 (P>0.05) for the short form and 4.04 +/- 0.8 vs 4.23 +/- 0. 6 (P>0.05) for the long form. The values for 72 h were 0.30 +/- 0.05 vs 0.24 +/- 0.05 (P>0.05) for the short form and 2.76 +/- 0.4 vs 5. 53 +/- 0.3 (P<0.01) for the long form respectively. CONCLUSION: The long form of the prolactin receptor is the dominant form at both time-points; however, the concentration of mRNA for this receptor isoform was specifically downregulated by prolactin treatment. Our results suggest that the short form of the prolactin receptor alone is unlikely to mediate the luteolytic action of prolactin, but that luteolytic events may be influenced via a change in the ratio of the two receptor isoforms.
Assuntos
Regulação da Expressão Gênica/efeitos dos fármacos , Luteólise/efeitos dos fármacos , Prolactina/farmacologia , RNA Mensageiro/metabolismo , Receptores da Prolactina/genética , 20-alfa-Di-Hidroprogesterona/sangue , Animais , Gonadotropina Coriônica/administração & dosagem , Gonadotropina Coriônica/farmacologia , Corpo Lúteo/anatomia & histologia , Feminino , Hipofisectomia , Tamanho do Órgão , Indução da Ovulação , Ratos , Ratos Sprague-Dawley , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
X-ray fluorescence and atomic absorption spectrometry were used to measure the concentrations of zinc, copper, and magnesium in the heart, liver, skeletal muscle, and kidney following isoproterenol-induced myocardial necrosis in male albino rats. Serum activities of lactic dehydrogenase (LDH), creatine phosphokinase (CPK), and glutamic oxaloacetic transaminase (SGOT) were also measured. There was depletion of myocardial zinc, copper, and magnesium on d 1, followed by an uptake of all these elements on d 2. The liver showed a significant uptake of magnesium, along with depletion of copper. There was no change in the kidney and skeletal muscle concentrations of these elements. Possible explanations for the observed changes and their therapeutic significance are presented.
RESUMO
X-ray fluorescence spectrometry and atomic absorption spectrometry were used in a quantitative study of zinc, copper, and magnesium in 71 postmortal human hearts. Samples were obtained from individuals who had demonstrated no previous clinical or subsequent pathological findings of myocardial infarction and from victims of a recent or an old infarction. A significant difference (p<0.001) in the elemental levels was observed between the noninfarct and the recent infarct groups. The noninfarct group had higher cardiac levels of all three elements. However, the difference in elemental concentrations between the noninfarct and the old-infarct groups was not significant. Cardiac levels of zinc (p<0.001) and copper (p<0.01) were significantly greater in the old-infarct group than in the recent-infarct group. Magnesium levels were higher in the recent-and-old-infarct group than in the recent infarct group (p<0.01). It is possible that the elements are redistributed during myocardial infarction, and that uptake of these elements (from the serum pool) by the heart may be important in maintaining myocardial integrity and function.
RESUMO
The muscles of the body are generally regarded as major targets for the actions of growth hormone (GH), but these tissues are difficult to utilize for in vitro studies of rapid cellular events. Therefore, the present study was conducted to determine whether the BC3H-1 cultured myocyte cell line possesses GH receptors and whether it exhibits metabolic responsiveness to GH. It was found that the myocytes possess receptors, which appear to be specific for GH and GH derivatives but which do not bind prolactin. The degree of GH binding observed was small, suggesting that receptors for GH are present in rather low abundance on these cells. To determine whether BC3H-1 myocytes are metabolically responsive to GH, the insulin-like activity of GH (i.e. the ability of GH to stimulate glucose metabolism) was examined, since this effect of GH is readily demonstrable when isolated rat muscle is incubated directly with the hormone. GH was found to stimulate glucose oxidation by the myocytes indicating that the receptors for GH on these cells are functional.
Assuntos
Hormônio do Crescimento/metabolismo , Músculos/metabolismo , Receptores da Somatotropina/análise , Animais , Ligação Competitiva , Linhagem Celular , Relação Dose-Resposta a Droga , Glucose/metabolismo , Hormônio do Crescimento/farmacologia , Camundongos , Músculos/efeitos dos fármacosRESUMO
The effects of isoacids, urea N, and S on ruminal fermentation of sugarcane bagasse- or corn stover-based diets were studied in sheep. Acetate production was taken as a measure of the fermentation rate. For the sugarcane bagasse diet, neither urea nor S supplementation changed ruminal acetate production. When N and S were combined, acetate production was 44% higher (3.16 vs. 2.18 mol/d). Similar effects were noted for the corn stover diet. Increasing the level of isoacids from .1 to .2 g/kg BW per d in the diet did not change acetate production for either diet. However, N supplementation of the sugarcane bagasse diet containing the low level of isoacids resulted in a 49% greater acetate production (2.86 vs. 1.91 mol/d). Acetate production was 90% higher (3.74 vs. 1.97 mol/d) when the diet containing the high level of isoacids was supplemented with N. The corresponding increases for corn stover were 12% (2.64 to 2.95 mol/d) and 35% (2.88 to 3.87 mol/d). The results suggest that NH3 N provided by the basal diet was more limiting than isoacids. Once the N deficiency was corrected, isoacids became limiting. Ruminal digestion of high fiber diets low in N was improved by supplementation with urea, isoacids, and S.
Assuntos
Fibras na Dieta/metabolismo , Rúmen/metabolismo , Ovinos/metabolismo , Acetatos/metabolismo , Animais , Butiratos/farmacologia , Fermentação , Rúmen/microbiologia , Enxofre/farmacologia , Ureia/farmacologia , Valeratos/farmacologiaRESUMO
Pituitary growth hormone (GH) functions physiologically to oppose the actions of insulin on carbohydrate and lipid metabolism by interfering with metabolic events that occur after insulin binds to its receptor. Which postreceptor effects are involved is presently unknown. Recently, we found that insulin rapidly stimulates a phosphatidylinositol phospholipase C (PI-PLC) in adipose tissue of obese (ob/ob) mice and that this effect of insulin is blocked by treatment of the animals with S-carboxymethylated human GH (RCM-hGH), a derivative having mainly anti-insulin activity. The activation of this PI-PLC by insulin is also inhibited by pertussis toxin. Thus, this study was performed to examine whether the inhibitory effect of GH on the activation of this PI-PLC is exerted at the level of signal transmission by guanine nucleotide binding proteins (G proteins). We found that the nonhydrolyzable GTP analogue, guanosine 5'-[gamma-thio]triphosphate, stimulated basal PI-PLC activity in plasma membranes of adipose tissue of saline-treated ob/ob mice, but it did not stimulate the enzyme in adipose membranes from RCM-hGH-treated mice. Also, RCM-hGH treatment markedly inhibited pertussis toxin-catalyzed ADP ribosylation of G protein alpha subunits in the membranes, suggesting some modification of the G proteins by GH. Immunoblot analysis of adipose membranes from saline- and RCM-hGH-treated mice using antiserum AS/7 (anti-Gi1 alpha and anti-Gi2 alpha) or antiserum EC/2 (anti-Gi3 alpha) showed no difference in the amount of Gi alpha-like protein between the groups. These findings suggest that GH interferes with the ability of a putative Gi-like protein to mediate the activation of PI-PLC in adipose membranes without altering the expression of the G protein.
Assuntos
Tecido Adiposo/enzimologia , Proteínas de Ligação ao GTP/fisiologia , Hormônio do Crescimento/análogos & derivados , Diester Fosfórico Hidrolases/metabolismo , Animais , Membrana Celular/enzimologia , Ativação Enzimática , Feminino , Hormônio do Crescimento/farmacologia , Nucleotídeos de Guanina/farmacologia , Guanosina 5'-O-(3-Tiotrifosfato)/farmacologia , Hormônios/farmacologia , Hormônio do Crescimento Humano , Cinética , Camundongos , Camundongos Obesos , Fosfatidilinositol Diacilglicerol-Liase , Fosfoinositídeo Fosfolipase C , Proteínas Recombinantes/farmacologia , Valores de ReferênciaRESUMO
In hypophysectomized rats, prolactin induces regression of the corpora lutea. Luteal regression is accompanied by infiltration of monocytes/macrophages, declines in luteal mass and plasma progestins, and increased staining for monocyte chemoattractant protein-1 (MCP-1). We investigated whether similar events are induced during the estrous cycle, after the proestrous prolactin surge. Rats were killed on proestrus or on estrus, and one ovary was frozen for immunohistochemical detection of MCP-1, monocytes/macrophages (ED1-positive), and differentiated macrophages (ED2-positive) and for in situ detection of apoptotic nuclei. Corpora lutea of the current (proestrus) or preceding (estrus) cycle were dissected from the ovaries of additional rats and frozen for the same analyses and for determination of total protein content. In sections of whole ovaries, intensity and distribution of MCP-1 staining were increased in corpora lutea of multiple ages on estrus as compared to proestrus, as were numbers of differentiated macrophages and apoptotic nuclei per high-power field. Sections of isolated corpora lutea showed these increases on estrus, and the number of monocytes/macrophages per high-power field was also significantly increased. Accompanying these inflammatory/immune events, the corpora lutea on estrus showed decreased weight and total protein per corpus luteum, as compared to corpora lutea on proestrus. These changes are consistent with a proposed role for prolactin in the initiation of luteal apoptosis and of a sequence of inflammatory/immune events that accompany regression of the rat corpus luteum during the normal estrous cycle.
Assuntos
Apoptose/fisiologia , Quimiocina CCL2/fisiologia , Luteólise/fisiologia , Macrófagos/fisiologia , Monócitos/fisiologia , Animais , Contagem de Células , Quimiocina CCL2/análise , Corpo Lúteo/química , Estro , Feminino , Imuno-Histoquímica , Macrófagos/citologia , Monócitos/citologia , Proestro , Ratos , Ratos Sprague-DawleyRESUMO
Fe, Ni, Cu and Zn were found by energy-dispersive X-ray fluorescence in calf thymus deoxyribonucleoprotein. The X-ray analyses indicated the absence of Cr, Mn and Co.
Assuntos
Desoxirribonucleoproteínas/análise , Metais/análise , Nucleoproteínas/análise , Animais , Bovinos , Cobre/análise , Ferro/análise , Níquel/análise , Timo/análise , Zinco/análiseRESUMO
In diabetes, increased oxidative stress, disruption of signal transduction pathways, and endothelial dysfunction have been critically implicated in the pathogenesis of experimental diabetic neuropathy (EDN). The development of nerve conduction slowing in diabetes is accompanied by depletion of the beta-amino acid taurine. Since taurine functions as an antioxidant, calcium modulator, and vasodilator, taurine depletion may provide a pathogenetic link between nerve metabolic, vascular, and functional deficits complicating diabetes. The mechanism(s) of nerve taurine depletion, the localization of critical taurine deficits, and its pathophysiological significance in EDN are however unknown. This study explored the pathophysiological effects of selective nerve taurine replacement in streptozotocin-diabetic (STZ-D) rats. A polyclonal human taurine transporter (TT) antibody was also generated in order to determine potential loci of critical taurine depletion. Two weeks of STZ-D reduced sciatic motor nerve conduction velocity (NCV) by 23% (P < 0.01), decreased composite nerve blood flow by 38% (P < 0.01), and reduced nerve taurine content by 29% (P < 0.05). In STZ-D rats, a 1% taurine diet corrected nerve taurine depletion, prevented motor NCV slowing, and partially attenuated composite nerve blood flow deficits. After 6 weeks of STZ-D, a 1% taurine diet ameliorated motor NCV slowing and endoneurial nutritive blood flow deficits, prevented digital sensory NCV slowing, and reduced ouabain-sensitive nerve (Na,K)-ATPase activity. Immunohistochemical studies localized taurine and the TT to the vascular endothelium and Schwann cells of the sciatic nerve. In conclusion, taurine depletion in the vascular endothelium and Schwann cells of the sciatic nerve may contribute to the neurovascular and metabolic deficits in EDN.
Assuntos
Proteínas de Transporte/metabolismo , Diabetes Mellitus Experimental/metabolismo , Neuropatias Diabéticas/metabolismo , Glicoproteínas de Membrana/metabolismo , Proteínas de Membrana Transportadoras , Condução Nervosa/fisiologia , Nervo Isquiático/metabolismo , Taurina/metabolismo , Animais , Antibacterianos , Velocidade do Fluxo Sanguíneo/efeitos dos fármacos , Velocidade do Fluxo Sanguíneo/fisiologia , Glicemia/efeitos dos fármacos , Glicemia/fisiologia , Pressão Sanguínea/efeitos dos fármacos , Pressão Sanguínea/fisiologia , Peso Corporal/efeitos dos fármacos , Peso Corporal/fisiologia , Diabetes Mellitus Experimental/fisiopatologia , Neuropatias Diabéticas/fisiopatologia , Endotélio Vascular/metabolismo , Humanos , Masculino , Condução Nervosa/efeitos dos fármacos , Coelhos , Ratos , Ratos Wistar , Nervo Isquiático/irrigação sanguínea , Nervo Isquiático/efeitos dos fármacos , Estreptozocina , Taurina/farmacologiaRESUMO
In previous studies ammonium salts of a mixture of isobutyrate, 2-methylbutyrate, isovalerate, and valerate were fed in a corn silage, corn, corn gluten meal, and urea diet to Holstein cows throughout lactation to define the optimum level of ammonium salts of milk production. The objective of this work was to conduct another dose response study using other forage and protein sources and to determine the effects of decreasing VFA intakes as lactation advanced. The concentrate portion of the diet contained 0, .4, .8, 1.2, or 1.6% ammonium salts of VFA. The forage to concentrate ratio was 50:50, 60:40, and 70:30 for the first, middle, and last third of lactation, respectively. The study was conducted at four university locations using 191 Holstein cows. Feeds used included corn silage, alfalfa silage or hay, corn, soybean meal, minerals, and vitamins. Treatment x location interactions were significant for milk yield during early lactation. During mid- and late lactation, supplemental VFA (.8%) improved milk and protein yield. Milk composition was not greatly affected by feeding VFA. In mid-lactation, cows fed .8% ammonium salts of VFA ate more feed than did controls. Feed efficiencies were similar among groups throughout the experiment. Cows fed VFA tended to gain less BW during lactation than did controls. Health and reproduction were not different among groups.
Assuntos
Bovinos/fisiologia , Ácidos Graxos Voláteis/farmacologia , Lactação/efeitos dos fármacos , Compostos de Amônio Quaternário/farmacologia , Ração Animal , Animais , Peso Corporal/efeitos dos fármacos , Relação Dose-Resposta a Droga , Ingestão de Alimentos/efeitos dos fármacos , Feminino , Leite/química , Leite/efeitos dos fármacos , Reprodução/efeitos dos fármacosRESUMO
We investigated the physiological basis for the trophic effect of glucocorticoids in rat corpora lutea in the absence of pituitary gonadotropins. Immature (Day 29) Sprague-Dawley rats were given eCG and hCG to induce the development of corpora lutea and were hypophysectomized on Day 32. Beginning on Day 40, rats received twice-daily s.c. injections of either dexamethasone (dex; 200 microg/rat/day) or vehicle (controls) and then were killed on Day 44. Plasma 20alpha-dihydroprogesterone, a major steroid produced by the corpora lutea, was higher (p = 0.01) in dex-treated than in control rats (44.5 +/- 2.3 vs. 23.0 +/- 5.6 ng/ml). Dexamethasone treatment increased lipid droplets and lipid in the corpora lutea as revealed by electron microscopy and oil red O staining. Cholesterol esters were higher in corpora lutea of dex-treated rats compared to controls (14.8 +/- 1.1 vs. 2.2 +/- 0.5 microg/mg corpora lutea wet tissue, respectively; p = 0.05). Another group of hypophysectomized rats was treated with either a high or a lower dosage of corticosterone, both of which caused an elevation to > 2-fold of plasma 20alpha-dihydroprogesterone concentration compared to controls. Glucocorticoid receptor protein (about 92 kDa) was detected in both luteal and nonluteal ovarian tissues in this animal model. These effects of glucocorticoids and the presence of the glucocorticoid receptor raise the possibility of a physiological role for glucocorticoids in the rat corpus luteum.