Ghrelin Expression Research Articles

Performance and serum parameters of calves (Bos taurus) subject to milk restriction associated with supplementation with 2-hydroxy-4-(methylthio)butanoic acid.

Our aim with this study was to evaluate the consumption, performance, quantitative characteristics of carcasses, biochemical profile, plasma levels of ghrelin and leptin, expression of the receptor for ghrelin (GHS-R1a) in the hypothalamus and duodenum, and the number of goblet cells in the duodenum of calves subjected to milk volume restriction and supplemented with 2-hydroxy-4-(methylthio)butanoic acid (HMTBa). We used 21 Holstein mixed-breed calves, aged between 3 and 15 d with an average weight of 36.8 kg, and housed in pens with troughs for hay, concentrate, and water. The study included two consecutive experimental periods (first period [P1] and second period [P2]) of 21 d each, with 7 d of adaptation to the diet and facilities. The calves were distributed in a completely randomized design in three treatments with seven repetitions. 1) Control: 6 liters of milk/d during P1 and 6 liters of milk/day during P2; 2) RES (milk restriction): 3 liters of milk/day during P1 and 6 liters of milk/day during P2; and 3) RES + HMTBa: 3 liters of milk/day during P1 and 6 liters of milk/day during P2 + 3.3 g of HMTBa/day in both periods. HMTBa was supplied in milk, and the amount of concentrated ration and hay provided and leftovers were recorded daily to estimate dry matter (DM) and crude protein consumption. Mean daily weight gain (DWG), final weight (FW), and feed conversion (FC) were obtained at the beginning and at the end of each 21-d period. Plasma concentrations of ghrelin and leptin, triglycerides, total protein, urea, lactate, creatinine, alkaline phosphatase, and cholesterol were measured for P1 and P2 at the end of each 21-d period. At the end of P2, animals were slaughtered; sections of the duodenum were collected to evaluate the expression of GHS-R1a and quantity of goblet cells; hypothalamus was used to evaluate the expression of GHS-R1a; rumen was used to evaluate the thickness of epithelium and keratin and the density, height, and width of ruminal papillae. In P1, total DM consumption, FW, DWG, glucose, and triglycerides were lower in the RES and RES + HMTBa groups (P < 0.001). In P2, there was an improvement in the FC of the RES + HMTBa group (compared with Control and RES groups) and a lower urea concentration in the RES group (compared with Control and RES + HMTBa groups) (P < 0.001). No differences were observed among groups regarding hormonal concentrations, histological parameters, and GHS-R1a expression in the duodenum and hypothalamus. Therefore, milk restriction combined with HMTBa supplementation promoted greater compensatory gain by a mechanism independent of changes in GHS-R1a expression and hormone levels of ghrelin and leptin.

Immunohistochemical analysis of ghrelin expression in various types of adrenal tumors.

Ghrelin, originally isolated from the endocrine cells of the gastric mucosa, is also expressed in many peripheral tissues, including normal adrenals and adrenocortical tumors. It was shown that ghrelin stimulates proliferation and inhibits apoptosis of adrenocortical cells. In the current study, we compared ghrelin expression at the protein level in various adrenal tumors. We analyzed whether immunoreactive ghrelin could be considered as a potential marker for different types of adrenal tumors. Study was carried out on 200 adrenal specimens arranged on microscope slide in tissue microarray format. We performed standardized immunohistochemical reactions with semiquantitative reaction intensity measurements. At the protein level, the expression of ghrelin was significantly reduced in adrenocortical adenocarcinoma in relation to the control group and pheochromocytoma as well as cancer-adjacent normal adrenal tissue. In contrast, a relatively high ghrelin expression was found in pheochromocytoma compared to all analyzed groups, with the exception of cancer-adjacent normal adrenal tissue. The ghrelin expression profile at the protein level may be associated with the type of adrenal tumor. In this context, our results suggest that adrenal immunoreactive ghrelin may be considered as a sensitive and specific marker for differentiating adrenocortical carcinoma from adrenocortical adenoma and pheochromocytoma.

Defective dystrophic thymus determines degenerative changes in skeletal muscle

In Duchenne muscular dystrophy (DMD), sarcolemma fragility and myofiber necrosis produce cellular debris that attract inflammatory cells. Macrophages and T-lymphocytes infiltrate muscles in response to damage-associated molecular pattern signalling and the release of TNF-α, TGF-β and interleukins prevent skeletal muscle improvement from the inflammation. This immunological scenario was extended by the discovery of a specific response to muscle antigens and a role for regulatory T cells (Tregs) in muscle regeneration. Normally, autoimmunity is avoided by autoreactive T-lymphocyte deletion within thymus, while in the periphery Tregs monitor effector T-cells escaping from central regulatory control. Here, we report impairment of thymus architecture of mdx mice together with decreased expression of ghrelin, autophagy dysfunction and AIRE down-regulation. Transplantation of dystrophic thymus in recipient nude mice determine the up-regulation of inflammatory/fibrotic markers, marked metabolic breakdown that leads to muscle atrophy and loss of force. These results indicate that involution of dystrophic thymus exacerbates muscular dystrophy by altering central immune tolerance.

Ghrelin Expression in Mast Cells of Infant Lung with Respiratory Distress Syndrome

Abstract This article sheds light on some features of ghrelin (GHR)- and tryptase (Try)-positive mast cells (MCs) distribution in human lung of preterm newborns with respiratory distress syndrome (RDS). GHR possessed anti-inflammatory activity and reliable therapeutic properties in some lung diseases. So far, GHR expression has been defined predominantly in neuroendocrine cells of bronchial mucosa in fetal and infant lungs. Lung tissue from 8 dead newborns with RDS were investigated immunohistochemically with anti-GHR and anti-Try antibodies. The number of GHR+ and Try+ MCs was determined in three locations –bronchi, bronchiole and in alveolar septa. MCs were more numerous around main bronchi with diminishing numbers around bronchiole and in alveolar septa. The number of MCs in the latter was increased in newborns with pneumonia. The number of GHR+ MCs in alveolar septa was lower in newborns with RDS as compared to newborns with RDS combined with pneumonia (2.83 ± 1.13 vs 4.81 ± 2.6, p &lt; 0.001). The amount of Try+ MCs along bronchial wall was significantly more than GHR+ MCs in RDS newborns (6.97 ± 4.53 vs 3.85 ± 4.30, p = 0.001). It could be supposed that pulmonary MCs increased in newborn lungs in inflammatory process. MCs in human lung contained GHR peptide that had immunomodulatory function and participated in hormone regulation of inflammation.

Dusk feeding in laying hens is shifted by light program via involvement of clock genes.

Dusk feeding is practised probably to satisfy the energy requirement during night. However, little is known on the changes with clock gene expressions during this feeding behaviour. In our present study, the linkage of clock gene expressions and feeding behaviour in dusk feeding was investigated in laying hens under two lighting programs: the conventional lighting program (Control) with a light period from 05:00 AM to 21:00 PM and a dark period from 21:00 PM to 05:00 AM; or the shifted lighting program group (SLP) with a light period from 02:00 AM to 18:00 PM and a dark period from 18:00 PM to 02:00 AM. The gene expression-related appetite and circadian rhythm were investigated in hypothalamus and proventriculus at 1, 3 and 5h before scotophase. The results demonstrated that dusk feeding was synchronously shifted with altered lighting program, dusk feeding was observed from 5h before scotophase in both groups. The expressions of anorexigenic gene proopiomelanocortin (POMC) and cocaine- and amphetamine-regulated transcript (CART) were downregulated (p<0.05) during dusk feeding and changed in pace with lighting program. The expressions of clock gene period 2 (Per2) and cryptochrome 1 (Cry1) in hypothalamus were downregulated (p<0.05) during dusk feeding and shifted by lighting program. In the proventriculus, ghrelin expression was decreased (p<0.05) during dusk feeding by lighting program. In conclusion, the expressions of clock genes Per2 and Cry1 are linked with the downregulated expressions of anorexigenic genes, POMC and CART, and in turn the augmented feed intake at dusk.

Ghrelin, Obestatin and Their Receptors As Well As Metabotropic Glutamate Receptor Assessment in Chronic Functional Constipation in Children.

The aim of this study was to examine the role of ghrelin, obestatin, and glutamate and their receptors in the pathogenesis of children functional constipation. Children ages 4-17 were the subject of the study: 121 children with constipation (55 boys and 66 girls), 36 patients of the same age (26 boys and 10 girls) were the controls. Expression of ghrelin, obestatin, and glutamate receptors on gastric and colon specimens taken by endoscopy were assessed. The concentration of the above agents was estimated in serum by the enzyme-linked immunosorbent assay test. The lower median serum concentrations of ghrelin, in the constipated children than in controls were confirmed (1.9 ng/mL vs 2.6 ng/mL, P < 0.05). The expression of the metabotropic receptor 7 for glutamate (mGlu7) RNA was higher in the stomach (32.49 vs 31.47, P < 0.05), and was lower in the rectum in constipated patients compared to the control group (31.76 vs 32.62, P < 0.05). A negative correlation between the concentration of ghrelin in serum and colonic transient time (P = 0.01, rho = -0.23) was shown in the study group.Higher median expression of obestatin receptor G protein-coupled receptor39 in rectal mucosae was found in a constipated group than in the controls (29.9 vs 26.9, P < 0.05). Ghrelin, and receptors for ghrelin, obestatin, and glutamate in gastrointestinal mucosa play a role in the pathogenesis of functional constipation in children.

53746 Body Composition and Metabolic Profiles in Infants of Diabetic Mothers (IDM) as Predictors of Hunger Signaling Gene Expression

ABSTRACT IMPACT: This study aims to advance the understanding of the biological mechanisms associated with feeding disturbances in infants born to diabetic mothers through non-invasive salivary gene expression analyses and body composition measurements at birth. OBJECTIVES/GOALS: To determine if non-invasive salivary gene expression analyses and body composition measurements at birth could elucidate biological mechanisms associated with aberrant feeding behaviors and disrupted metabolic profiles commonly seen in infants born to diabetic mothers. METHODS/STUDY POPULATION: This prospective cohort study enrolls subjects born at ≥35 weeks’ gestation without a history of intrauterine growth restriction or major congenital anomalies. The diabetic cohort is defined as infants born to mothers with gestational diabetes or type 2 diabetes. The primary outcome is salivary expression of the hunger signaling genes, AMPK and NPY2R. Secondary outcomes include infant body composition measurements, obtained by skinfold measurement and/or air displacement plethysmography, and salivary expression of the adipokines, leptin, ghrelin, and adiponectin. Multiple logistic regression will be used to determine which factors are associated with AMPK and NPY2R expression. RESULTS/ANTICIPATED RESULTS: We propose that poor oral intake seen in infants of diabetic mothers may be due to alterations in the expression of hunger signaling genes (decreased expression of AMPK; increased expression of NPY2R) resulting in a diminished feeding drive in these large for gestational age infants. In addition, infant adiposity and the expression of genes involved in the adipoinsular axis will be inversely proportional to feeding volume intake. Namely, increased neonatal fat mass will be associated with increased expression of leptin and decreased expression of ghrelin and adiponectin. DISCUSSION/SIGNIFICANCE OF FINDINGS: Infants of diabetic mothers are at higher risk of poor oral feeding in the newborn period. This study aims to elucidate the link between neonatal body composition, adipoinsular axis, and hunger signaling to explain this unique feeding phenotype.

A White Grape Juice Extract Reduces Fat Accumulation through the Modulation of Ghrelin and Leptin Expression in an In Vivo Model of Overfed Zebrafish.

A caloric surplus and a sedentary lifestyle are undoubtedly known to be the leading causes of obesity. Natural products represent valuable allies to face this problematic issue. This study was planned to assess the effect of a white grape (Vitis vinifera) juice extract (WGJe) in diet-induced obese zebrafish (Danio rerio). Fish were divided into four different diet groups: (i) normally fed (NF); (ii) overfed (OF); (iii) WGJe-supplemented NF (5 mL/L in fish water); (iv) WGJe-supplemented OF. Body mass index (BMI) was extrapolated each week. After the fourth week, euthanized zebrafish were processed for both microscopic evaluations and gene expression analyses. OF zebrafish showed higher BMI values with respect to NF counterparts, an effect that was hindered by WGJe treatment. Moreover, histological analyses showed that the area of the adipose tissue, as well as the number, size, and density of adipocytes was significantly higher in OF fish. On the other hand, WGJe was able to avoid these outcomes both at the subcutaneous and visceral levels, albeit to different extents. At the gene level, WGJe restored the altered levels of ghrelin and leptin of OF fish both in gut and brain. Overall, our results support the anti-obesity property of WGJe, suggesting its potential role in weight management.

Selenium nanoparticle and selenomethionine as feed additives: effects on growth performance, hepatic enzymes' activity, mucosal immune parameters, liver histology, and appetite-related gene transcript in goldfish (Carassius auratus).

The aim of this study was to assess the effects of different dietary selenium sources, selenium nanoparticle (nSe), and selenomethionine (SeMet) as feed additives on growth performance, hepatic enzymes' activity, biochemical, mucosal immune parameters, liver histology, and appetite-related gene transcript in goldfish (Carassius auratus). At first, goldfish juveniles (n=480; mean 4.54 g) were fed dietary selenium nanoparticle at 0, 0.3, 0.6, and 0.9 mg nSe/kg diet and SeMet at 0, 0.3, 0.6, and 0.9 mg Se/kg for 9 weeks. Growth performance was evaluated using standard procedures. Blood, skin mucus, and tissue samples (liver and intestine) were collected for biochemical, mucosal immune response, histology, and ghrelin and insulin-like growth factor-I (IGF-I) gene expression. The results showed that fish fed diets fortified with 0.6 mg nSe/kg and 0.6 mg Se/kg had a significant higher weight gain, specific growth rates (SGR), and lower feed conversion ratios (FCR) than fish fed basal diets (p<0.05). Furthermore, dietary nSe and SeMet enhanced blood biochemical profiles especially alkaline phosphatase (ALP) (p < 0.05) and mucosal immunity than the control group in goldfish. Moreover, the liver histological investigation showed that fish fed 0.9 mg of SeMet and nSe kg-1 diets had higher liver lesion scores such as karyolysis, lipidosis, and hyperemia while fish fed 0, 0.3, and 0.6 mg of SeMet and nSe kg-1 diets had small liver changes at 9 weeks. The study further established that inclusion of nSe and SeMet in the diet of goldfish greatly promoted ghrelin and IGF-1genes expressions (p <0.05). Overall, dietary nSe performs better than SeMet and basal diets. The results evoked that nSe and SeMet stimulate the growth, biochemical, and mucosal immunity in goldfish at 0.6 mg/kg.

Prenatal music exposure influences weight, ghrelin expression, and morphology of rat hypothalamic neuron cultures.

Music plays an important role in brain physiology, in some areas related to emotions, food intake and body weight, such as the hypothalamus. There are different frequencies to which it can be tuned, today the most utilized is at 440Hz, while in the past the 432Hz frequency was more used to show particular effects on brain. It is known that Ghrelin, a peptide hormone, regulates food intake in the hypothalamus; in a previous paper, we reported that musical stimuli at 432Hz modified the Ghrelin expression in the rat, increasing beneficial effects on metabolism. In this study, we used this frequency and we focused our attention on body weight, Ghrelin expression, and neuron morphology in hypothalamic cultures. To investigate the role of music, we utilized newborn pups from pregnant rats that were exposed to music stimuli at 432Hz during the perinatal period and for the postnatal period, some for 3days (P3) and others for 6days (P6). Some pups were not exposed to music stimuli (controls). Our results showed that music increased the body weight of pups; in addition, enhanced Ghrelin expression in hypothalamic neurons and their axonal elongation were highlighted by immunocytochemical techniques. Moreover, we found that the positive music effect started in pups at P3 and increased at P6 compared with controls. These results suggest that the musical frequency at 432Hz could stimulate the orexigenic Ghrelin effects influencing the increase in body weight and affecting the number of hypothalamic neurons expressing Ghrelin.

Tryptase- and ghrelin positive mast cells in the interalveolar septa of rat’s lung

The mast cell mediators and distribution of lung mast cells in rats are often discussed in experimental studies on pulmonary fibrotic and allergic processes associated with changes in numbers of these cells, but information on the normal distribution of metachromatic and tryptase-positive mast cells in the interalveolar septa is scarce. There are no data on the presence of ghrelin in lung mast cells as well as the age-specific features of localisation and the number of mast cells in the interalveolar septa in rats of different ages. Therefore, the purpose of the present study was to determine the distribution of metachromatic, tryptase-, and ghrelin-positive mast cells in the interalveolar septa in 20 day-, 3 month- and 1 year-old rats. Tissue sections stained with toluidine blue had been taken from the left lung to visualise metachromasia and immunohistochemical expression of tryptase and ghrelin. The results showed that the amount of metachromatic mast cells in the interalveolar septa was significantly lower than that of tryptase- and ghrelin-positive cells. This allowed suggesting that mast cells were permanent occupants of the rat lung parenchyma and, on the other hand, the expression of ghrelin in their granules was most likely related to the synthesis of this protein. Our study showed that immunohistochemical identification by tryptase expression was more accurate than toluidine blue staining.

Chronic stress accelerates the process of gastric precancerous lesions in rats.

Background: Gastrointestinal cancers account for 20% of all deaths worldwide. Gastric cancer (GC) patients are susceptible to psychological change, especially depression which is commonly induced by chronic stress. Gastric precancerous lesions (GPL) is an important prodromal stage in the occurrence of gastric cancer. Chronic stress influences the prognosis of GC and may influence the process of GPL as well.Methods: Sixty SD rats were randomly divided into a control group, GPL group, and GPL+CUMS group. In the GPL group, 200μg/mL N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) free drinking method combined with intermittent fasting was applied to establish the GPL animal model. Based on this, we combined the GPL rats with chronic unpredicted mild stress (CUMS) to establish a comprehensive model. We then evaluated their behavior by open field tests and sucrose preference tests. We tested the IL-6, IL-10, TNF-α, Ghrelin, Leptin and Somatostatin (SS) levels in serum and observed the expression of Ghrelin and Gastrokine 2(GKN2) in the gastric mucosa of rats with tumors by immunofluorescence.Results: Our results showed that GPL and GPL+CUMS rats all displayed a significantly decreased total distance and mean velocity traveled in the open field test. The percentages of sucrose preference were significantly decreased in the GPL+CUMS group compared to the control group. In addition, IL-6 and TNF-α were significantly increased in both the GPL and GPL+CUMS groups. Furthermore, the GPL+CUMS group showed significantly increased TNF-α levels in serum compared to the GPL rats. Our results showed that the expression of NF-κB, p53, and BCL-2 were significantly increased while BAX was reduced in the GPL and GPL+CUMS groups. Moreover, Ghrelin and Leptin levels in serum were significantly decreased in the GPL and GPL+CUMS groups. SS levels in serum were significantly increased in the GPL+CUMS group. Additionally, we found that the GPL+CUMS rats with tumors not only had strong expression of GKN2 on the luminal side and the lamina propria of the gastric mucosa and tumor, but also had expression of Ghrelin on the luminal side of the gastric mucosa. The areas that showed strong expression of GKN2 and Ghrelin, are all located around the blood vessels in the tumor.Conclusions: GPL rats under chronic stress would aggravate the conditions of GPL, shorten the process of GPL, and increase the risk of tumorigenesis. In addition, the close monitoring of the mental health of cancer survivors and precancerous lesion patients is suggested to be of great significance in the prevention and treatment of cancer.

Ghrelin gastric tissue expression in patients with morbid obesity and type 2 diabetes submitted to laparoscopic sleeve gastrectomy: immunohistochemical and biochemical study.

Obesity and type 2 diabetes mellitus (T2DM) are leading causes of morbidity and mortality worldwide. Ghrelin is implicated in the pathophysiology of both disease states. Laparoscopic sleeve gastrectomy is an emerging safe therapeutic technique for patients with morbid obesity. Since the removal of ghrelin-secreting cells by sleeve gastrectomy may be associated with diminished hunger sensation the aim of the study was to: (i) compare body weight and body mass index (BMI) in both obese non-diabetic and obese diabetic patient groups, (ii) determine the ghrelin expression in the resected gastric tissue in both groups, (iii) evaluate relationships between ghrelin cell expression and pre- and post-operative serum ghrelin concentration and glucose levels, and (iv) assess the influence of sleeve gastrectomy on serum glycaemic parameters in this patient population. Twenty morbidly obese female patients from Saudi Arabia, of whom ten suffered from T2DM participated in the study. All subjects underwent laparoscopic sleeve gastrectomy. The removed fundus, body and antrum were biopsied and underwent immunohistochemical staining to detect ghrelin cell expression. Serum samples were assayed for ghrelin concentration and indicators of glycaemic status at the baseline and three months after sleeve gastrectomy. BMI (p < 0.05) and body weight (p < 0.001) were significantly lower in non-diabetic obese patients compared with diabetic patients before and 3 months after the surgery. Also, pre-operative serum ghrelin level was higher in non-diabetic patients compared with diabetic patients group, and postoperative plasma ghrelin level was reduced in diabetic patients (p < 0.001) compared with non-diabetic patients. Gastric fundic mucosa of the diabetic patients exhibited lower number of ghrelin-positive cells (p < 0.05) compared with non-diabetic patients. There were significant negative correlations between pre- and post-operative ghrelin serum level and blood glucose (r = -0.736, p = 0.0002 and r = -0.656, p = 0.0007, respectively) in all patient populations. The results of this study suggest that the diabetic status of obese female patients may affect the incidence of ghrelin cells in three major stomach's regions and this novel observation warrants further studies.

Intestinal dysmotility after bowel resection in rats is associated with decreased ghrelin and vimentin expression and loss of intestinal cells of Cajal.

This study provides novel insight into the mechanisms of intestinal dysmotility following massive small bowel resection. We show that 2 wk after bowel resection in rats, impaired intestinal motility was associated with loss of interstitial cells of Cajal (ICC; downregulation of transmembrane member 16A (TMEM16A) and c-kit expression) as well as with decreased vimentin, desmin, and ghrelin levels. Impaired intestinal motility led to a decrease in final body weight, suggesting less effective nutrient absorption. The purpose of this study was to evaluate the mechanisms of intestinal motility in a rat model of short bowel syndrome (SBS). Rats were divided into three groups: Sham rats underwent bowel transection; SBS-NSI rats underwent a 75% bowel resection and presented with normal intestinal size (NSI) at euthanasia and hypermotility patterns; SBS-DYS showed dysmotile (DYS) enlarged intestine and inhibited motility patterns. Animals were euthanized after 2 wk. Illumina's digital gene expression (DGE) analysis was used to determine the intestinal motility-related gene expression profiling in mucosal samples. Intestinal motility-related and ICC genes and protein expression in intestinal muscle layer were determined using real-time PCR, Western blotting, and immunohistochemistry. Gastrointestinal tract motility was studied by microcomputer tomography. From 10 Ca2+ signaling pathway-related genes, six genes in jejunum and seven genes in ileum were downregulated in SBS vs. Sham animals. Downregulation of TMEM16A mRNA and protein was confirmed by real-time PCR. Rapid intestinal transit time in SBS-NSI rats correlated with a mild decrease in TMEM16A, c-kit, and vimentin mRNA and protein expression (vs/. Sham animals). SBS-DYS rats demonstrated enlarged intestinal loops and delayed small intestinal emptying (on imaging studies) that were correlated with marked downregulation in TMEM16A, c-kit, vimentin, and ghrelin mRNA and protein levels compared with the other two groups. In conclusion, 2 wk following massive bowel resection in rats, impaired intestinal motility was associated with decreased vimentin and ghrelin gene and protein levels as well as loss of ICC (c-kit and TMEM16A).NEW & NOTEWORTHY This study provides novel insight into the mechanisms of intestinal dysmotility following massive small bowel resection. We show that 2 weeks after bowel resection in rats, impaired intestinal motility was associated with loss of interstitial cells of Cajal (downregulation of TMEM 16A, and c-kit expression) as well as with decreased vimentin, desmin, and ghrelin levels. Impaired intestinal motility led to decrease in final body weight, suggesting less effective nutrient absorption.

RETRACTED: Protective effect of naringin on small intestine injury in NSAIDs related enteropathy by regulating ghrelin/GHS-R signaling pathway

To investigate the mechanism of Ghrelin/GHS-R signaling pathway in small intestine injury induced by NSAIDs related enteropathy. To clarify the mechanism network of intestinal mucosal repair with naringin as a new therapeutic method. Naringin was used as the intervention method, observed the damage of small intestinal mucosa and detected the expression of ghrelin, GHS-R, leptin and TNF-α by electron microscopy, HE staining and immunohistochemistry. Compared with the control group, the weight of rats in the model group decreased, the thickness of intestinal mucosa became thinner, the structure of intestinal mucosa changed, the expression of ghrelin, GHS-R and leptin decreased, the expression of TNF-α increased. Compared with the model group, the intestinal mucosa of the treatment group was repaired, the expression of ghrelin, GHS-R and leptin was increased, and the expression TNF-α was decreased. The mechanism of intestinal mucosal damage in patients with NSAIDs related enteropathy may be related to the decreased expression of ghrelin, GHS-R and leptin, and promotion of TNF-α secretion. Naringin can effectively promote the secretion of ghrelin and leptin, the expression of GSH-R, and inhibit the release of TNF-α, so as to repair intestinal mucosa naringin will become a new method to treat and prevent NSAIDs related intestinal diseases.

Gastric ghrelin cells in obese patients are hyperactive.

Distribution and activity of ghrelin cells in the stomach of obese subjects are controversial. We examined samples from stomachs removed by sleeve gastrectomy in 49 obese subjects (normoglycemic, hyperglycemic and diabetic) and quantified the density of ghrelin/chromogranin endocrine cells by immunohistochemistry. Data were compared with those from 13 lean subjects evaluated by gastroscopy. In 44 cases (11 controls and 33 obese patients) a gene expression analysis of ghrelin and its activating enzyme ghrelin O-acyl transferase (GOAT) was performed. In 21 cases (4 controls and 17 obese patients) the protein levels of unacylated and acylated-ghrelin were measured by ELISA tests. In 18 cases (4 controls and 14 obese patients) the morphology of ghrelin-producing cells was evaluated by electron microscopy. The obese group, either considered as total population or divided into subgroups, did not show any significant difference in ghrelin cell density when compared with control subjects. Inter-glandular smooth muscle fibres were increased in obese patients. In line with a positive trend of the desacylated form found by ELISA, Ghrelin and GOAT mRNA expression in obese patients was significantly increased. The unique ghrelin cell ultrastructure was maintained in all obese groups. In the hyperglycemic obese patients, the higher ghrelin expression matched with ultrastructural signs of endocrine hyperactivity, including expanded rough endoplasmic reticulum and reduced density, size and electron-density of endocrine granules. A positive correlation between ghrelin gene expression and glycemic values, body mass index and GOAT was also found. All obese patients with type 2 diabetes recovered from diabetes at follow-up after 5 months with a 16.5% of weight loss. Given the known inhibitory role on insulin secretion of ghrelin, these results suggest a possible role for gastric ghrelin overproduction in the complex architecture that takes part in the pathogenesis of type 2 diabetes.

Ghrelin inhibited pressure overload–induced cardiac hypertrophy by promoting autophagy via CaMKK/AMPK signaling pathway

Ghrelin, a novel gut hormone, has been shown to exert protective effects on cardiac dysfunction and remodeling. However, the underlying mechanisms of its protective effects remain unclear. Here, we investigated the effects of ghrelin on cardiac hypertrophy and explored the mechanisms involved. Ghrelin (30 μg.kg−1. day−1) was systemically administered to rats with cardiac hypertrophy induced by abdominal aortic constriction (AAC) by a mini-osmotic pump the next day after surgery continuously for 4 weeks. The AAC treated rats without ghrelin infusion showed decreased ghrelin content and expression of its receptors in the hearts. Exogenous ghrelin greatly attenuated cardiac hypertrophy as shown by heart weight to tibial length (HW/TL), hemodynamics, echocardiography, histological analyses, and expression of hypertrophic markers induced by AAC. This corresponded with decreased cardiac fibrosis and inflammation in the hearts of AAC rats treated with ghrelin. Moreover, ghrelin significantly increased the myocardial expression of autophagy markers, which was further confirmed in cultured cardiomyocytes. Concurrently, cardiomyocyte apoptosis in vivo and in vitro was ameliorated by ghrelin, which was reversed by inhibition of autophagy. The enhancement of autophagy and inhibition of apoptosis by ghrelin were eliminated on pretreatment with compound C, an AMP-activated protein kinase (AMPK) inhibitor. Furthermore, inhibition of Ca2+/Calmodulin-dependent protein kinase kinase (CaMKK), an upstream kinase of AMPK, made ghrelin fail to activate AMPK and simultaneously reversed ghrelin’s promotion of autophagy. In conclusion, ghrelin could exert its cardioprotective effects on cardiac hypertrophy by promoting autophagy, possibly via CaMKK/AMPK signaling pathway.

Effect ofClostridium on proliferating cell nuclear antigen and ghrelin in the small intestine of fattening pigs fed with deoxynivalenol

Grains and feed are severely contaminated by deoxynivalenol (DON) globally, threatening both human and animal health. Research on bio-degradation of DON, in general, is gaining attention. The aim of this research was to estimate the effect ofClostridium sp. WJ06 as a microbiological detoxification of DON based on the expression and distribution of proliferating cell nuclear antigen (PCNA) as well as ghrelin in the small intestine. A total of 24 fattening pigs were randomly divided into three groups. The control group was fed with a basic diet, the DON group was fed with DON at 5.0 mg/kg in feed, and the DON+C group was provided DON feed withClostridium sp. WJ06. Several selected blood parameters, the intestinal morphology, and the expression and distribution of PCNA and ghrelin, were evaluated. The results proved that the selected blood parameters were altered, the intestinal villi were damaged, the epithelium was shed, as well as the expression and distribution of PCNA and ghrelin were changed by DON exposure. These toxic effects were prevented by the addition ofClostridium sp. WJ06. In short, the addition ofClostridium sp. WJ06 to the feed may eliminate the toxic effects of DON in fattening pigs. An underlying mechanism is likely modulation of the expression and distribution of PCNA and ghrelin.

Effects of Hot-Melt Extruded Nano-Copper as an Alternative for the Pharmacological Dose of Copper Sulfate in Weanling Pigs.

This study was conducted to investigate the effects of hot-melt extrusion (HME)-processed copper (Cu) sulfate supplementation on the growth performance, gut microbiota, metabolic function of Cu, and bioavailability of Cu in weanling pigs fed a corn-soybean meal basal diets. A total of 180 piglets (Yorkshire × Landrace × Duroc) of mixed-sex randomly were allotted to six treatments on the basis of initial average body weight (6.36 ± 0.39kg) to six dietary treatments. There were six replicates in each treatment with 5 pigs per replicates. The dietary treatments included levels of CuSO4 (IN6, 6mg Cu/kg diets; IN125, 125mg Cu/kg diets), nano-CuSO4 (HME6, 6mg Cu/kg diets; HME65, 65mg Cu/kg diets; and HME125, 125mg Cu/kg diets), and Cu-methionine (ORG125, 125mg Cu/kg diets). The weanling pigs fed diets supplemented with the HME65 and HME125 showed a greater body weight and feed intake compared with IN6 and IN125 (P < 0.05). The weaning pigs fed diets supplemented with the HME125 showed the highest digestibility of gross energy in phase 1 and phase 2 (P < 0.05). The supplementation of HME125 significantly reduced theEscherichia coli (E.coli) in cecum and colon (P < 0.05). The supplementation of HME65 showed statistically equivalent effect on reduction of E. coli in the cecum and colon compared with IN125 and ORG125 treatments. The villus height in duodenum and jejunum of piglets in HME65 and HME125 treatments were higher than ORG125, HME6, IN6, and IN125 (P < 0.05). The gene expression of Atox1 was upregulated in IN125, HME125, and ORG125 treatments (P < 0.05). The expression of Sod1 was increased in IN125 treatment compared with IN6 treatment (P < 0.05). The HME125 treatment had the highest gene expression of ghrelin (P < 0.05). The Cu concentration of serum and liver was higher in the HME125 treatment than the HME6, IN6, and IN125 treatments (P < 0.05). The HME125 and ORG125 treatments showed a lower fecal Cu compared with IN125 treatment (P < 0.05). Taken together, these results suggest that the HME65 can be an alternative to IN125 in weanling pigs due to the greater overall average daily gain, improved villus height, and higher bioavailability.

PREDICTIVE VALUE OF FOLLICULAR FLUID GHRELIN IN WOMEN WITH POLYCYSTIC OVARIAN DISEASE WHO UNDERGO INTRACYTOPLASMIC SPERM INJECTION

Ghrelin’s role in reproduction was first suggested by its wide expression in many human reproductive tissues including its immunohistochemical expression in the human ovary. Ghrelin expression is identified in the seminiferous tubuli of the interstitial Leyding and Sertoli cells in male testicles, whereas it is identified in the ovarian hilus interstitial cells and mature corpus lutea in females.It is postulated that ghrelin plays a local regulatory role in the male and female gonads. In females, Ghrelin acts centrally by suppressing hypothalamic GnRH release and GnRH-induced gonadotropin secretion by the pituitary; and in gonads it acts by increasing luteolytic factors such as PGF2 alfa, and ghrelin inhibiting estradiol and progesterone biosynthesis in granulose-lutein cells as well as granulosa cell (GC) proliferation.This potential regulatory effect on steroidogenesis and ovarian cellular function led to the hypothesis that ghrelin may mediate the relationship between metabolism and oocyte quality in addition to possible role in the regulation of embryo development in as the cleavage development before the six-cell stage of a preimplantation human embryo depends solely on maternally inherited material. Ghrelin levels can, therefore, be an indicator of an underlying pathology as in humans in states of chronic malnutrition such as anorexia nervosa, serum ghrelin levels may be increased, while in other conditions it could be associated with decreased ghrelin levels such as obesity and polycystic ovarian syndrome (PCOS).