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Role of leptin in the control of postprandial pancreatic enzyme secretion

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Leptin released by adipocytes has been implicated in the control of food intake but recent detection of specific leptin receptors in the pancreas suggests that this peptide may also play some role in the modulation of pancreatic function. This study was undertaken to examine the effect of exogenous leptin on pancreatic enzyme secretion in vitro using isolated pancreatic acini, or in vivo in conscious rats with chronic pancreatic fistulae. Leptin plasma level was measured by radioimmunoassay following leptin administration to the animals. Intraperitoneal (i.p.) administration of leptin (0.1, 1, 5, 10, 20 or 50 µg/kg), failed to affect significantly basal secretion of pancreatic protein, but markedly reduced that stimulated by feeding. The strongest inhibition has been observed at dose of 10 µg/kg of leptin. Under basal conditions plasma leptin level averaged about 0.15 ± 0.04 ng/ml and was increased by feeding up to 1.8 ± 0.4 ng/ml. Administration of leptin dose-dependently augmented this plasma leptin level, reaching about 0.65 ± 0.04 ng/ml at dose of 10 µg/kg of leptin. This dose of leptin completely abolished increase of pancreatic protein output produced by ordinary feeding, sham feeding or by diversion of pancreatic juice to the exterior. Leptin (10-10-10-7 M) also dose-dependently attenuated caerulein-induced amylase release from isolated pancreatic acini, whereas basal enzyme secretion was unaffected. We conclude that leptin could take a part in the inhibition of postprandial pancreatic secretion and this effect could be related, at least in part, to the direct action of this peptide on pancreatic acini
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Role of endothelin-1-dependent up-regulation of leptin in oral mucosal repair

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Leptin, a multifunctional hormone that regulates food intake and energy expenditure, has emerged recently as an important modulator of inflammatory cascades associated with wound healing. In this study, we applied the animal model of buccal mucosal ulcer to investigate the role of endothelin-1 (ET-1) and leptin in soft oral tissue repair. Using groups of rats with experimentally induced buccal mucosal ulcers we show that ulcer onset was characterized by a marked increase in the mucosal level of ET-1 and leptin. However, while the ET-1 level gradually declined with healing, the mucosal level of leptin increased reaching maximum expression on the 4th day of healing. Therapeutic administration of phosphoramidon, an inhibitor of ECE-1 activity, not only led to a 53.2% drop in the ET-1, but also produced a dose-dependent reduction (up to 50.9%) in the mucosal level of leptin and up to 42.3% decline in the rate of ulcer healing. A marked drop (54.2%) in the mucosal level of leptin and the reduction (46.8%) in the rate of ulcer healing was also attained in the presence of ETA receptor antagonist BQ610 administration, but not the ETB receptor antagonist BQ788. Moreover, administration of ERK inhibitor, PD98059 in the presence of ETB receptor antagonist, but not the ETA receptor antagonist, caused the reduction the mucosal leptin level as well as a decline in the rate of ulcer healing. Our findings are the first to implicate the requirement for both ET-1 and leptin in orderly progression of the events of soft oral tissue repair. We also show that ET-1 is a key factor in up-regulation of leptin production associated with oral mucosal ulcer healing , and that the effect of ET-1 on leptin production is a consequence of ETA receptor activation and subsequent signaling through MAPK/ERK.
The age-related inverse relationship between gene expression of lipogenic enzymes and leptin gene expression as well as inhibitory effect of leptin on lipogenic enzyme’s gene expression suggests that leptin could be responsible in part for the low rate of lipogenesis in white adipose (WAT) of old rats. Based on the data published recently we propose a model for the direct inhibitory effect of leptin on lipogenesis. This model may explain the age-related decrease of lipogenic activity in WAT. It is likely that despite of higher concentration of noradrenaline (which inhibits leptin gene expression in WAT) in old animals, the age-dependent decrease of b-adrenergic receptor density in rat adipocytes may lead to the increase of leptin gene expression and to the increase of WAT leptin concentration. High concentration of leptin in adipose tissue decreases sterol regulatory element binding protein–1 (SREBP-1) gene expression by paracrine and/or autocrine action on adipocyte, which leads to the decrease of SREBP-1c level (mature form). The suppression of SREBP-1c synthesis causes a decrease of lipogenic enzyme’s gene expression which consequently results in lower rate of fatty acid synthesis in WAT. This model does not exclude the indirect, via hypothalamus (by decreasing food consumption), inhibitory action of leptin on WAT lipogenesis. Therefore, it is likely that leptin exerts its inhibitory effect on WAT lipogenesis both directly at the level of adipocytes, and indirectly through hypothalamus by decreasing food intake. The inhibitory effect of high leptin concentration on lipogenesis in WAT of old rats could prevent over-accumulation of triacylglycerols in adipocyte, and by this way could protect against further development of the fat mass.
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Leptin does not seem to influence glucose uptake by bovine mammary explants

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Leptin, a protein produced and secreted by adipocytes, is know to regulate food intake and whole-body energy metabolism, but knowledge about its possible effect in bovine mammary gland is scarce. Leptin may be involved in the regulation of glucose transport even though this effect at the tissue level remains controversial. Once uptaken by the mammary gland, glucose is utilised in several ways but the majority, about 60-70%, is drained for lactose synthesis. This study was aimed at investigating the effect of leptin on glucose regulation in bovine mammary gland. We have examined the effects of leptin on the expression of GLUT1 mRNA, pyruvate kinase (PK) as well as glucose-6-phosphate dehydrogenase (G6PDH) activity. Treatment of mammary gland explants with recombinant leptin did not influence glucose assimilation, pathway transport (GLUT1 mRNA) and glucose metabolism (PK and G6PDH) in this tissue. The results from this study seem to exclude an involvement of leptin in glucose uptake and metabolism in bovine mammary gland.
We examined the effect of leptin on renal function and renal Na+,K+-ATPase and ouabain-sensitive H+,K+-ATPase activities in the rat. Leptin was infused under general anaesthesia into the abdominal aorta proximally to the renal arteries. Leptin infused at doses of 1 and 10 µg/kg/min increased urine output by 40% and 140%, respectively. Urinary Na+ excretion increased in rats receiving leptin at doses of 0.1, 1, and 10 µg/kg/min by 57.6%, 124.2% and 163.6%, respectively. Leptin had no effect on creatinine clearance, potassium excretion and phosphate excretion. Na+,K+-ATPase activity in the renal medulla of rats treated with 1 and 10 µg/kg/min leptin was lower than in control animals by 25.5% and 33.2%, respectively. In contrast, cortical Na+,K+-ATPase as well as either cortical or medullary ouabain-sensitive H+,K+-ATPase activities did not differ between leptin-treated and control animals. The effect of leptin on Na+,K+-ATPase activity was abolished by actin depolymerizing agents, cytochalazin D and latrunculin B, and by phosphatidylinositol 3-kinase (PI3K) inhibitors, wortmannin and LY294002. These results indicate that: 1) natriuretic effect of leptin is mediated, at least in part, by decrease in renal medullary Na+,K+-ATPase activity, 2) inhibition of medullary Na+,K+-ATPase by leptin is mediated by PI3K and requires integrity of actin cytoskeleton.
Leptin is a multifunctional regulator in numerous tissues, including the pituitary. It is not known, whether the porcine pituitary is a source of leptin synthesis and possesses the leptin receptor protein. It is also unknown, if a relationship exists between expression levels of these proteins in the pituitary and physiological status of sows. Therefore, the aim of the study was 1] to examine, by Western-blotting analysis, the expression levels of leptin and the long form of leptin receptor (OB-Rb) in the porcine anterior (AP) and posterior (NP) pituitary gland during mid- and late-luteal phases of the oestrous cycle (days 10 - 12 and 14 - 16) as well as during two stages of early pregnancy (days 14 - 16 and 30 - 32); and 2] to localise, using in situ hybridisation method (ISH), the expression of leptin and OB-Rb genes in the pituitary gland in the above mentioned stages of the cycle and pregnancy. Western-blotting analysis showed that leptin protein expression in AP was higher in the late-luteal phase than in the mid-luteal phase, while OB-Rb protein expression in both lobes was higher in the mid-luteal phase. In turn, during pregnancy leptin protein content in AP and OB-Rb protein content in NP were more pronounced on days 14 - 16 than on days 30 - 32. Comparison of leptin and OB-Rb protein expression levels in AP between the mid-luteal phase and two periods of pregnancy showed, respectively, stimulation of leptin protein and inhibition of OB-Rb protein expressions during both examined stages of pregnancy. Taking AP from late-luteal phase as the point of reference, it was revealed stimulation of leptin expression during earlier period of pregnancy, whereas on days 30 - 32 of pregnancy both the hormone and its receptor expressions were diminished. In turn, comparison of leptin and OB-Rb protein expression levels in NP between the late-luteal phase and days 14 - 16 or 30 - 32 of pregnancy showed inhibition of leptin protein expression and stimulation of OB-Rb protein expression during pregnancy. Moreover, ISH studies localised leptin and OB-Rb mRNAs expression in the cells of AP as well as NP tissue during the two stages of the cycle and pregnancy. In conclusion, our findings suggest that leptin is produced within the pituitary in the pig and may participate in auto/paracrine manner in the regulation of this gland function during the luteal phase of the oestrous cycle and early pregnancy.
Leptin interplays with other peptides to control feeding behaviour in humans and animals. Using exendin-4, an agonist of glucagon-like peptide-1, we investigated whether leptin modifies its effect on food intake in the rat. In the first series, exendin-4 alone (0.1, 2 or 10 µg per rat), leptin alone (0.1, 2, 10 or 100 µg per rat) or exendin-4 and leptin together (0.1 + 0.1, 2 + 2, 10 + 10, or 2 + 100 µg per rat, respectively) were injected once intraperitoneally. In the second series animals were injected either with exendin-4 (2 µg) alone, leptin (10 µg) alone, or leptin (10 µg) + exendin-4 (2 µg) daily for 5 subsequent days. At the lowest dose used, leptin and exendin-4 injected once together, but not separately, reduced significantly a 24-hour food intake. When used in higher doses, however, leptin did not change the exendin-4-dependent suppressory effect on food consumption. No significant differences in food intake were seen between rats treated repeatedly with exendin-4 only and animals injected with both drugs. Hence, leptin and exendin-4 may act additively to inhibit appetite when present in low concentrations while, at high leptin doses, this effect is abolished. The lack of synergistic effects of exendin-4 and high leptin concentrations on food intake may explain, at least in part, mechanisms responsible for leptin resistance in subjects with hyperleptinaemia.
Leptin is a polypeptide that plays a key role in the regulation of energy homeostasis and is also linked, among others, to mechanisms controlling reproductive processes. Data concerning the involvement of leptin in controlling reproductive functions at the level of hypothalamus and pituitary in the pig are limited. Therefore, in the present study, an expression of genes coding for leptin and long-form leptin receptor (Ob-Rb) was determined by a semiquantitative reverse transcription polymerase chain reaction (RT-PCR) in the discrete areas of porcine hypothalamus (medial basal hypothalamus - MBH, preoptic area - POA, stalk median eminence - SME) and pituitary (anterior - AP and posterior/neural - NP parts) during the luteal phase of the cycle (days 10-12 and 14-16) and two early stages of pregnancy (days 14-16 and 30-32). Leptin gene expression in MBH was found to be higher in the mid- than in the late-luteal phase, whereas in other structures studied it remained unchanged during these periods. More pronounced differences were noted in expression of Ob-Rb gene, which was increased in MBH, AP and NP during the late-luteal phase in comparison to the mid-luteal one, whilst the relationship in the POA was reversed. In turn, during pregnancy, leptin gene expression in all tested areas of hypothalamus as well as Ob-Rb mRNA content in MBH were higher on days 30-32 than on days 14-16. In contrast, in the anterior pituitary, Ob-Rb gene expression was more pronounced on days 14-16 than during later stage of pregnancy. Comparison of leptin and Ob-Rb mRNA content in studied structures between the mid-luteal phase and days 14-16 of pregnancy revealed inhibition of leptin gene expression in almost all examined tissues (MBH, POA, SME, NP) during early pregnancy whereas Ob-Rb gene expression was inhibited in POA but stimulated in both parts of the pituitary during this stage. In summary, obtained results suggest an involvement of leptin in the regulation of hypothalamic-pituitary axis activity during both the luteal phase of the cycle and early pregnancy in pigs.
The secretion of gonadotrophins from anterior pituitary cells can be modulated by leptin and signals originating from the immune system, among others, by nitric oxide (NO). There are some studies that have demonstrated a role for leptin and NO in the regulation of FSH in rodents, however, no similar data are available in regard to ewes. Therefore, the objective of the present study was to analyse the leptin effect on GnRH-induced FSH secretion from the ovine anterior pituitary cells in vitro. Additionally, the influence of leptin on NO release and its role in the GnRH and leptin–modulated secretion of FSH from pituitary gland of ewes was investigated. The obtained results show that the influence of leptin on FSH secretion is biphasic. Leptin in concentration 10-8 and 10-7 M/l significantly enhances, whereas 10-6 and 10-5 M/l of leptin suppresses FSH secretion from the pituitary cells in comparison to the control. The secretion of FSH and NO release under the influence of leptin are in very high positive correlation (r=0.77). The inhibition of NO synthesis with L-NAME, instead, disables leptin from the stimulation of FSH secretion.
Activation of cytosolic phospholipase A2 (cPLA2) by bacterial LPS for the rapid release of arachidonic acid from membrane phospholipids is considered a key step in the generation of platelet-activating factor (PAF), recognized as the most proximal mediator of inflammatory events triggered by bacterial infection. In this study, we report on the role of leptin in modulation of the detrimental consequences of H. pylori LPS-induced cPLA2 activation that result in the disturbances in gastric mucin synthesis. Employing gastric mucosal cells labeled with [3H] arachidonic acid, we show that H. pylori LPS-induced cPLA2 activation, associated with up-regulation in apoptosis and PAF generation, and the impairment in gastric mucin synthesis, was subject to a dose-dependent suppression by leptin, as well as the inhibition by MAFP, a specific inhibitor of cPLA2. A potentiation in the countering capacity of leptin on the LPS-induced up-regulation in apoptosis, arachidonic acid release and PAF generation was attained in the presence of ERK inhibitor, PD98059, while PI3K inhibitor, wortmannin had no effect. On the other hand, the prevention by leptin of the LPS detrimental effect on mucin synthesis was subject to suppression by wortmannin, an inhibitor of PI3K as well as the inhibitor of ERK, PD98059. Moreover, potentiation in the effect of leptin on the LPS-induced decrease in mucin synthesis was attained with cPLA2 inhibitor, MAFP as well as PAF receptor antagonist, BN52020. The results of our findings point to H. pylori LPS-induced ERK-dependent cPLA2 activation as a critical factor influencing the level of PAF generation, and hence the extent of pathological consequences of H. pylori infection on the synthesis of gastric mucin. Furthermore, we show that leptin counters the pathological consequences of H. pylori-induced cPLA2 activation on gastric mucin synthesis through the involvement in signaling events controlled by MAPK/ERK and PI3K pathways.
The aim of the study was to investigate the effect of different dietary fats (sunflower oil, fish oil, lard) and their level in a diet (10% and 20% wt/wt), as well as different time of experimental diets administration (3 or 6 weeks) on plasma leptin level in Wistar rats. The type of dietary fat influenced significantly the plasma leptin concentration which was higher in groups receiving sunflower oil in a diet and lower in fish oil groups. There was a positive correlation between plasma leptin level and PUFA intake (r=0.371, p<0.003) and linoleic acid intake and its plasma level (r=0.462, p<0.0002 and r=0.381, p<0.005, respectively) as well as a negative correlation between plasma leptin and intake of docosahexaenoic acid and its plasma level (r=-0.360, p<0.004 and r=-0.360, p<0.008, respectively). There was no effect of the level of fat in a diet and time of experimental diets administration on plasma leptin, except of fish oil groups, where the concentration of the hormone decreased with an increasing level of this fat in a diet. It can be concluded that a change in the composition of dietary fatty acids can alter leptin production in adipose tissue.
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Brain-gut axis and its role in the control of food intake

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Gastrointestinal tract (GIT) and nervous system, both central (CNS) and enteric (ENS), are involved in two-way extrinsic communication by parasympathetic and sympathetic nerves, each comprising efferents fibers such as cholinergic and noradrenergic , respectively, and afferent sensory fibers required for gut-brain signaling. Afferent nerves are equipped with numerous sensors at their terminals in the gut related to visceral mechano- chemo- and noci-receptors, whose excitations may trigger a variety of visceral reflexes regulating GIT functions, including the appetitive behaviour. Food intake depends upon various influences from the CNS as well as from the body energy stores (adipocytes) that express and release the product of Ob gene, leptin, in proportion to fat stored and acting in long-term regulation of food intake. Leptin acts through receptors (Ob-R) present in afferent visceral nerves and hypothalamic arcuate nucleus (ARC), whose neurons are capable of expressing and releasing neuropeptide Y (NPY) and agouti related protein (AgRP) that activate the ingestive behaviour through paraventricular nucleus (PVN) (“feeding center”). In addition, to this long-term regulation, a short-term regulation, on meal-to-meal basis, is secured by several gut hormones, such as cholecystokinin (CCK), peptides YY (PYY) and oxyntomodulin (OXM), released from the endocrine intestinal cells and acting via G-protein coupled receptors (GPCR) either on afferent nerves or directly on ARC neurons, which in turn inhibit expression and release of food-intake stimulating NPY and AgRP, thereby inducing satiety through inhibition of PVN. In contrast, during fasting, the GIT, especially oxyntic mucosa, expresses and releases appetite stimulating (orexigenic) factors such as ghrelin and orexins (OX) -A and OX-B, and cannabinoid CB1 agonist. Ghrelin activates growth-hormone secretagogue receptor (GHS-R) in hypothalamic ARC and stimulates growth hormone (GH) release and in vagal afferents to promote the expression and release of hypothalamic NPY and AgRP stimulating PVN and driving ingestive behaviour. The balance and interaction between anorexigenic (CCK, PYY, OXM) and orexigenic (ghrelin and OX) factors originating from GIT appears to play an important role in short-term regulation of food intake and growth hormone (GH) release. An impairment of this balance may result in disorders of feeding behaviour and weight gain (obesity) or weight loss (cachexia).
Five single nucleotide polymorphisms in leptin and leptin receptor genes were analysed for their possible impact on estimating breeding values for somatic cell count score (SCS) in milk, longevity and reproductive traits. Used were 309 active Polish Holstein-Friesian bulls. The LEP-C(-963)T,LEP-Y7F, LEP-R25C, LEP-A80V, and LEPR-T945M genotypes were identified using the PCRRFLP method. For linked leptin mutations, the additional haplotype analysis was performed. The results obtained suggest that three polymorphisms of bovine LEP gene may be associated with nonreturn rate in cows. The most significant effect was found for LEP-A80V. Moreover, the LEPRT945M mutation seemed to be related to the age at first insemination.
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Neuroendocrine control of metabolic homeostasis in Polish centenarians

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Neuropeptides play a pivotal role in the control of metabolic homeostasis. We aimed to evaluate the release of neuropeptides involved in the control of energy homeostasis in relation to metabolic status in aging humans. The study group consisted of 183 women: 75 centenarians (above 100 yrs old), 26 elderly women (below 70 yrs), 45 younger women (mean 26 yrs) and 37 obese women (mean 41.6 yrs). Fasting plasma concentration of leptin, adiponectin, ghrelin active, neuropeptide Y (NPY) and insulin were measured. Our results showed several differences in the metabolic and neurohormonal status in the centenarian group. The incidence of hypertension, glucose intolerance, insulin resistance and dyslipidemia was lower compared with obese women. Leptin and NPY concentrations were significantly lower than in elderly and obese subjects. Moreover, NPY level was higher than that in the younger group. Plasma adiponectin values were higher than in any of the other group. Insulin levels were significantly lower compared with the young and obese groups. Furthermore, a negative correlation was found between adiponectin and HOMA-IR, and adiponectin and insulin. Ghrelin active concentrations were significantly lower compared with the young subjects. However, ghrelin levels were higher than in obese subjects. We conclude that altered neuropeptide activity in centenarians may play a role in the mechanisms contributing to prolonged survival.
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