BACKGROUND

<em>Nesfatin</em>-<em>1</em> is a recently discovered neuropeptide derived from its precursor nucleobindin-2 (NUCB2) and has been implicated in the regulation of feeding and energy metabolism. It is located in the brain and also produced at the periphery and present in the plasma. However, its pathophysiological role in humans remains unknown. Polycystic ovary syndrome (PCOS) is commonly presented with obesity, insulin resistance, hyperandrogenemia and hirsutism.

OBJECTIVE

To characterize serum nesfatin-<em>1</em> levels in PCOS women and determine association of nesfatin-<em>1</em> with metabolic parameters.

METHODS

It is a cross-sectional study of 55 PCOS and 28 healthy women matched in age, in a university hospital setting. Anthropometric, hormonal, metabolic parameters and nesfatin-<em>1</em> blood levels were determined.

RESULTS

<em>Nesfatin</em>-<em>1</em> levels were significantly higher in PCOS group compared with the controls 37<em>1</em>.43 ± 2.50 versus 275.55 ± <em>1</em>.74 pg/mL. Multivariate logistic regression analysis that contains: nesfatin-<em>1</em>, body mass index and homeostasis model assessment index revealed significant correlation of nesfatin-<em>1</em> with the existence of PCOS (p < 0.05).

CONCLUSIONS

Higher nesfatin-<em>1</em> levels in PCOS women compared to control group may suggest a possibility that nesfatin-<em>1</em> may play some role in the PCOS.

<em>Nesfatin</em>-<em>1</em> is a recently identified 82 amino acid peptide shown to have an anorexigenic effect on rodents when administrered centrally and peripherally. <em>Nesfatin</em>-<em>1</em> is expressed not only in neurones of various brain areas, including the hypothalamic and brainstem nuclei, but also in peripheral organs, such as the stomach and the pancreas. <em>Nesfatin</em>ergic neurones were reported to participate in the regulation of satiety signals and in the responses to other stimuli, including restraint stress, abdominal surgery, and lipopolysaccharide-induced inflammation. The present study aimed to investigate whether NUCB2/<em>nesfatin</em>-<em>1</em> expressing neurones also take part in the central signalling activated in response to hypoglycaemia and therefore are involved in central glucose sensing. Using immunolabelling methods based on the detection of the neuronal activation marker c-Fos and of <em>nesfatin</em>-<em>1</em>, we showed that peripheral injection of insulin induced a strong activation of <em>nesfatin</em>-<em>1</em>-expressing neurones in the brain vagal-regulatory nuclei, including the arcuate nucleus, paraventricular nucleus, lateral hypothalamic area, dorsal motor nucleus of the vagus (DMNX) and nucleus of the tractus solitarius. In response to intracellular glucopaenia induced by i.p. or i.c.v. 2-deoxyglucose injection, the c-Fos/<em>nesfatin</em>-<em>1</em> colocalisations observed at the hypothalamic and brainstem levels were similar to those observed after insulin-induced hypoglycaemia. Moreover, using Fluorogold as a retrograde tracer, we showed that <em>nesfatin</em>ergic preganglionic DMNX neurones activated by hypoglycaemia target the stomach and the pancreas. Taken together, these results suggest that a subpopulation of <em>nesfatin</em>ergic neurones belongs to the central network activated by hypoglycaemia, and that <em>nesfatin</em>-<em>1</em> participates in the triggering of physiological and hormonal counter-regulations observed in response to hypoglycaemia.

<em>Nesfatin</em>-<em>1</em>, a recently discovered satiety molecule, is localized in neurons of the hypothalamus and brain stem and colocalized with stress-related substances. However, the relation between <em>nesfatin</em>-<em>1</em> and stressor related behaviors like anxiety and/or fear has not yet been investigated in human subjects. In the present study, our aim was to investigate whether there was a relationship between plasma <em>nesfatin</em>-<em>1</em> levels and generalized anxiety disorder. The study group consisted of 40 patients (BMI, 22.98 ± 0.56) with generalized anxiety disorder and 34 age-matched healthy male control subjects (23.05 ± 0.4). Patients fully met the fourth Diagnostic and Statistical Manual of Mental Disorders, text revision. Blood samples for <em>nesfatin</em>-<em>1</em> were drawn at the end of an overnight fasting period at least <em>1</em>0h. Plasma <em>nesfatin</em>-<em>1</em> levels were measured and found significantly lower in anxiety disorder group than in control group (0.35 ± 0.037 ng/ml vs. 0.63 ± 0.080 ng/ml, respectively, p<0.05). Low <em>nesfatin</em>-<em>1</em> levels may be related with generalized anxiety disorder.

The aim of this study was to clarify the hypothalamic neuropeptides that are associated with hyperphagic feeding in Tsumura Suzuki Obese Diabetes (TSOD) mice, a model of type 2 diabetes with polygenic abnormalities. TSOD mice showed an increase in body weight and hyperleptinemia from <em>1</em> month of age and hyperphagic feeding, hyperglycemia, hyperlipidemia and hyperinsulinemia from 3 to <em>1</em>2 months of age compared with age-matched non-diabetic control Tsumura Suzuki Non Obesity (TSNO) mice. The mRNA level of nucleobindin-2 (NUCB2), the precursor of the anorexigenic neuropeptide <em>nesfatin</em>-<em>1</em>, was significantly decreased in the hypothalamus of TSOD mice compared with that in TSNO mice from 3 to <em>1</em>2 months of age. The protein level of NUCB2 was significantly decreased in the hypothalamus of TSOD mice compared with that in TSNO mice at 3 months of age. The mRNA levels of galanin, melanin-concentrating hormone, neuropeptide Y, and pro-opiomelanocortin were significantly changed in the hypothalamus in TSOD mice at several time points. Another model of type 2 diabetes, db/db mice, which is a mutant mouse that lacks a functional leptin receptor, showed hyperphagic feeding but no change in hypothalamic NUCB2 mRNA compared with non-diabetic control db/+ mice. The results suggest that the disrupted control of hypothalamic NUCB2-mediated signaling may contribute to hyperphagic feeding in TSOD mice. In addition, the mechanism for the development of hyperphagic feeding in TSOD mice is different than that in db/db mice.

Deoxynivalenol (DON), produced by the cereal-contaminating Fusarium fungi, is a major trichothecene responsible for mycotoxicoses in farm animals, including swine. The main effect of DON-intoxication is food intake reduction and the consequent body weight loss. The present study aimed to identify brain structures activated during DON intoxication in pigs. To this goal, we used c-Fos staining which constitutes a useful approach to identify activated neurons. We showed that per os administration of Fusarium graminearum extracts (containing the equivalent of <em>1</em>mg DON per kg of body weight) induced an increase in c-Fos immunoreactivity in several central structures, including the ventrolateral medulla (VLM), dorsal vagal complex (DVC), paraventricular nucleus of the hypothalamus (PVN), arcuate nucleus (Arc), supraoptic nucleus (SON) and amygdala (CeA). Moreover, we coupled c-Fos staining with phenotypic markers detection in order to specify the neuronal populations activated during DON intoxication. This phenotypic characterization revealed the activation of catecholaminergic but not of serotoninergic neurons in response to the toxin. In this context, we also paid a particular attention to NUCB2/<em>nesfatin</em>-<em>1</em> positive cells, since <em>nesfatin</em>-<em>1</em> is known to exert a satiety effect. We report here, for the first time in the pig brain, the presence of NUCB2/<em>nesfatin</em>-<em>1</em> neurons in the VLM, DVC, PVN, Arc and SON, and their activation during DON intoxication. Taken together, these data show that DON stimulates the main structures involved in food intake in pigs and suggest that catecholaminergic and NUCB2/<em>nesfatin</em>-<em>1</em> neurons could contribute in the anorexigenic effects of the mycotoxin.

Biological fluids (milk and serum/plasma) and cheese whey milk-derived fluid contain numerous molecules, especially amino acids and proteins. Therefore, the purpose of this study was to find out whether cheese whey (n:6), cow milk (n:6) and its blood (n=6) have adropin, <em>nesfatin</em>-<em>1</em>, apelin-<em>1</em>2, ghrelins and salusin peptides. Adropin, <em>nesfatin</em>-<em>1</em>, apelin-<em>1</em>2 concentrations were measured by ELISA, whereas ghrelin and salusin concentrations were measured by EIA methods. It was found that adropin, <em>nesfatin</em>-<em>1</em>, apelin-<em>1</em>2, des-acylated ghrelin and salusins in cheese whey were higher than in the corresponding milk peptides and plasma of dairy cows, with the exception of salusin alpha and acylated ghrelin in milk being the same than that of the corresponding cheese whey concentration and plasma of dairy cows. A correlation was also found between milk peptides and cheese whey, as also with plasma of dairy cows. The data suggest that peptides in cow milk might be an important and nutritious food for (neonatal) calves and human diet due to their biological and physiological properties.

<em>Nesfatin</em>-<em>1</em>, corticotropin-releasing hormone (CRH), thyrotropin-releasing hormone (TRH), and hypothalamic neuronal histamine act as anorexigenics in the hypothalamus. We examined interactions among <em>nesfatin</em>-<em>1</em>, CRH, TRH, and histamine in the regulation of feeding behavior in rodents. We investigated whether the anorectic effect of <em>nesfatin</em>-<em>1</em>, α-fluoromethyl histidine (FMH; a specific suicide inhibitor of histidine decarboxylase that depletes hypothalamic neuronal histamine), a CRH antagonist, or anti-TRH antibody affects the anorectic effect of <em>nesfatin</em>-<em>1</em>, whether <em>nesfatin</em>-<em>1</em> increases CRH and TRH contents and histamine turnover in the hypothalamus, and whether histamine increases <em>nesfatin</em>-<em>1</em> content in the hypothalamus. We also investigated whether <em>nesfatin</em>-<em>1</em> decreases food intake in mice with targeted disruption of the histamine H<em>1</em> receptor (H<em>1</em>KO mice) and if the H<em>1</em> receptor (H<em>1</em>-R) co-localizes in <em>nesfatin</em>-<em>1</em> neurons. <em>Nesfatin</em>-<em>1</em>-suppressed feeding was partially attenuated in rats administered with FMH, a CRH antagonist, or anti-TRH antibody, and in H<em>1</em>KO mice. <em>Nesfatin</em>-<em>1</em> increased CRH and TRH levels and histamine turnover, whereas histamine increased <em>nesfatin</em>-<em>1</em> in the hypothalamus. Immunohistochemical analysis revealed H<em>1</em>-R expression on <em>nesfatin</em>-<em>1</em> neurons in the paraventricular nucleus of the hypothalamus. These results indicate that CRH, TRH, and hypothalamic neuronal histamine mediate the suppressive effects of <em>nesfatin</em>-<em>1</em> on feeding behavior.

We recently identified a novel satiety peptide, <em>nesfatin</em>-<em>1</em>, containing 82 amino acids derived from the precursor peptide, nucleobindin 2 (NUCB2), from a troglitazone (TZ)-induced cDNA library. We examined the molecular mechanism underlying TZ-induced NUCB2 mRNA expression. Although TZ induced the mRNA expression in HTB<em>1</em>85 cells, a nuclear run-on assay revealed no significant change in the transcription of the gene. Surprisingly, HTB<em>1</em>85 cells possessed no functional peroxisome proliferator-activated receptor-gamma. We therefore examined the effect of TZ on the mRNA's stability. The half-life of NUCB2 mRNA was approximately 6 h, and incubation with TZ increased this to 27 h. Furthermore, this increase was completely inhibited by an ERK inhibitor, PD98059, and phosphorylated ERK<em>1</em>/2 was significantly increased after 30 min incubation with TZ. In addition, we cloned the entire NUCB2 gene and identified four adenylate/uridylate-rich elements (AREs) in the 3' untranslated region (UTR), to which several proteins of HTB<em>1</em>85 extracts treated with TZ bound. The reporter assay fused with 3'UTR showed that the second and third AREs were crucial. Furthermore, the human NUCB2 gene spanned 55 kb and contained <em>1</em>4 exons and <em>1</em>3 introns. The transcriptional start site formed clusters around 246 bp upstream from the translational start site. We confirmed that a construct containing 5889 bp of the promoter region was very active in neuron-derived cell lines but not stimulated by TZ. These findings demonstrated a novel action of derivatives of thiazolidinediones, oral insulin-sensitizing antidiabetic agents, to stabilize the mRNA of NUCB2 through AREs in the 3'UTR by activating the ERK<em>1</em>/2 pathway independently of peroxisome proliferator-activated receptor-gamma.

<em>Nesfatin</em>-<em>1</em>, discovered by Oh-I and his coworkers in 2006, is a multi-functional peptide hormone with an approximate MW of 9.8 kDa and a half-life of 23.5 minutes. This peptide is found in three different forms, <em>nesfatin</em>-<em>1</em>, <em>nesfatin</em>-2 and <em>nesfatin</em>-3, all three of which are formed from the precursor NUCB2 by proteolytic processing. The 30-amino acid middle segment of <em>nesfatin</em>-<em>1</em> (M30) is responsible for limiting food intake, while the exact physiological role of <em>nesfatin</em>-2 and <em>nesfatin</em>-3 is unknown. This review will focus on <em>nesfatin</em>-<em>1</em> in relation with tissue and fluid distribution, considerations for its analysis in body fluids, and its potential as a biomarker for some diseases.

<em>Nesfatin</em>-<em>1</em> is a naturally occurring, 82-amino acid peptide processed from the precursor nucleobindin 2 (NUCB2), a highly conserved protein among vertebrates. In fish, two isoforms of NUCB2 (NUCB2A and NUCB2B) exist, and <em>nesfatin</em>-<em>1</em> has been identified in goldfish and Ya fish. We recently reported the presence and appetite suppressing effects of <em>nesfatin</em>-<em>1</em> in goldfish. The main objectives of this study were to characterize NUCB2 in zebrafish, and determine whether NUCB2 mRNAs are affected by food availability. Tissue distribution of NUCB2A and NUCB2B mRNAs, and NUCB2/<em>nesfatin</em>-<em>1</em>-like immunoreactivity (ir) in the gut of zebrafish were also investigated. In zebrafish, <em>nesfatin</em>-<em>1</em> region (<em>1</em>-82 amino acids) in NUCB2A is 78% identical to NUCB2B. Both NUCB2A and NUCB2 mRNAs were most abundant in the liver, while less expression was found in other tissues including the brain and gut. NUCB2/<em>nesfatin</em>-<em>1</em>-like immunoreactivity was detected in the mucosal layer cells of zebrafish anterior gastrointestinal tract. NUCB2A and NUCB2B mRNA expression were decreased in the brain of zebrafish 3h after feeding, and after a 7-day food deprivation. Both NUCB2A and NUCB2B mRNAs in the gut were also decreased following 7 days of food deprivation, while NUCB2B mRNA was increased in the liver. Our results provide molecular and functional evidences to support potential anorectic and metabolic roles for endogenous <em>nesfatin</em>-<em>1</em> in zebrafish. To our knowledge, this is the first report on NUCB2B characterization in vertebrates.

Lifestyle-related diseases are associated with overeating and lack of exercise. The purpose of this study was to investigate the effect of exercise and high-fat diet on plasma adiponectin and <em>nesfatin</em> levels. Mice were housed for 4 weeks in 4 groups, which included the non-exercise and normal diet (SN), exercise and normal diet (EN), non-exercise and high-fat diet (SF) and the exercise and high-fat diet (EF) group. The mice in the exercise groups were housed in cages with a running wheel and were subjected to voluntary exercise. The food intake (Kcal) of the mice in the exercise groups increased compared to that of the mice in the non-exercise groups (P<0.0<em>1</em>). Body weight and visceral fat decreased in the mice in the EF group compared to the mice in the SF group (P<0.0<em>1</em> and P<0.05). The temperature of the mice in the EF group increased compared to that of the mice in the SN group (P<0.05). Blood glucose, insulin (P<0.0<em>1</em>), cholesterol (P<0.0<em>1</em>) and triglyceride concentrations (P<0.0<em>1</em>) increased in the SF group compared to the normal diet groups. Furthermore, plasma insulin and cholesterol concentrations increased in the SF group compared to the exercise groups (P<0.0<em>1</em>). Plasma adiponectin and <em>nesfatin</em>-<em>1</em> levels in the SF group decreased compared to the SN group (P<0.05). Exercise under a high-fat diet antagonized the significant decrease in the <em>nesfatin</em>-<em>1</em> level. Exercise together with a high-fat diet affected the plasma levels of adiponectin and <em>nesfatin</em>. It is therefore suggested that exercise together with a high-fat diet can affect various diseases via adiponectin and <em>nesfatin</em>.

Some gastrointestinal and pancreatic hormones are potently secreted by meal intake and reduce food intake, therefore these hormones play a role in the meal-evoked satiety peptides. Previous reports have demonstrated that peripheral administration of these gastrointestinal or pancreatic hormones decrease feeding and the anorectic effects are abolished by lesions of vagal afferent nerves using surgical or chemical protocols, indicative of the involvement of the vagal afferents. Vagal afferent nerves link between several peripheral organs and the nucleus tractus solitarius of the brainstem. The present review focuses on cholecystokinin, peptide YY(3-36), pancreatic polypeptide, and <em>nesfatin</em>-<em>1</em> released from endocrine cells of the gut and pancreas. These hormonal peptides directly act on and increase cytosolic Ca(2+) in vagal afferent nodose ganglion neurons and finally suppress food intake via vagal afferents. Therefore, peripheral terminals of vagal afferents could sense gastrointestinal and pancreatic hormones and regulate food intake. Here, we review how the vagal afferent neurons sense a variety of gastrointestinal and pancreatic hormones and discuss its physiological significance in regulation of feeding.

<em>Nesfatin</em>-<em>1</em>, the N-terminal fragment of nucleobindin 2 (NUCB2), is an 82 amino-acid peptide that inhibits food intake and exerts weight-reducing effects. <em>Nesfatin</em>-<em>1</em> has been proposed as a potential anti-obesity peptide. However, studies to date have mainly focused on the acute satiety effects of centrally administered <em>nesfatin</em>-<em>1</em>. The main objective of our studies was to characterize the long-term/chronic effects of peripheral administration of <em>nesfatin</em>-<em>1</em> on whole-body energy balance and metabolic partitioning in male Fischer 344 rats. Short-term (<em>1</em> day) subcutaneous infusion of <em>nesfatin</em>-<em>1</em> (50 μg/kg body weight/day) using osmotic mini-pumps increased spontaneous physical activity and whole-body fat oxidation during the dark phase. This was accompanied by decreased food intake and basal metabolic rate compared to saline infused controls. On the seventh day of <em>nesfatin</em>-<em>1</em> infusion, cumulative food intake, and total spontaneous physical activity during the dark phase were significantly reduced and elevated, respectively. Meanwhile, intraperitoneal injection of <em>nesfatin</em>-<em>1</em> only caused a dark phase specific reduction in food intake and an increase in physical activity. NUCB2 mRNA expression in the brain and stomach, as well as serum NUCB2 concentrations were significantly reduced after 24 h fasting, while a post-prandial increase in serum NUCB2 was found in ad libitum fed rats. Collectively, our results indicate that chronic peripheral administration of <em>nesfatin</em>-<em>1</em> at the dose tested, results in a sustained reduction in food intake and modulation of whole body energy homeostasis.

Signals from the hypothalamus govern food intake and energy balance. A new study describes <em>nesfatin</em>-<em>1</em>, a hypothalamic and brainstem peptide whose expression decreases during fasting. Although central treatment with <em>nesfatin</em>-<em>1</em> inhibited food intake and <em>nesfatin</em>-<em>1</em> blockade increased food intake, the role and mechanism of <em>nesfatin</em> in energy balance remains unclear.

<em>Nesfatin</em>-<em>1</em> is a novel anorexigenic hormone which has close relationship with diabetes, obese, anorexia nervosa, psychiatric disorders and neurogenic diseases. The aim of our study was to evaluate levels of plasma <em>nesfatin</em>-<em>1</em> among patients presenting with coronary artery disease and the correlation between <em>nesfatin</em>-<em>1</em> levels and other clinical parameters. Fasting plasma levels of <em>nesfatin</em>-<em>1</em> were tested in 48 acute myocardial infarction (AMI) patients, 74 stable angina pectoris (SAP) patients and 34 control subjects. All of them were examined by coronary angiography. The severity of coronary atherosclerosis was assessed using the Gensini score. Plasma <em>nesfatin</em>-<em>1</em> levels were significantly lower in AMI group than SAP group or control group (0.9<em>1</em>±0.08 ng/mL vs. 0.98±0.<em>1</em>9 ng/mL and <em>1</em>.09±0.39 ng/mL, respectively, P<0.05). In AMI patients, plasma <em>nesfatin</em>-<em>1</em> levels were negatively correlated with high-sensitivity C-reactive protein, neutrophil% or Gensini scores. Such information implies that lower <em>nesfatin</em>-<em>1</em> concentration may play a very important role in the development of AMI.

In the present study, we explored whether dietary lipid content influences the gut microbiome in adult zebrafish. Diets containing three different lipid levels (high [HFD], medium [MFD], and low [LFD]) were administered with or without the supplementation of Lactobacillus rhamnosus (P) to zebrafish in order to explore how the dietary lipid content may influence the gut microbiome. Dietary lipid content shifted the gut microbiome structure. The addition of L. rhamnosus in the diets, induced transcriptional reduction of orexigenic genes, upregulation of anorexigenic genes, and transcriptional decrease of genes involved in cholesterol and triglyceride (TAG) metabolism, concomitantly with lower content of cholesterol and TAG. Probiotic feeding also decreased <em>nesfatin</em>-<em>1</em> peptide in HFD-P and attenuated weight gain in HFD-P and MFD-P fed zebrafish, but not in LFD-P group. Intestinal ultrastructure was not affected by dietary fat level or probiotic inclusion. In conclusion, these findings underline the role of fat content in the diet in altering gut microbiota community by shifting phylotype composition and highlight the potential of probiotics to attenuate high-fat diet-related metabolic disorder.

BACKGROUND

Early prediction of disease severity in acute pancreatitis (AP) is crucial. The aim of this study was to investigate the body-mass index (BMI), plasma leptin, <em>nesfatin</em>-<em>1</em> and ghrelin levels as potential markers predicting peripancreatic necrosis and severity in acute pancreatitis.

METHODS

In the study period, 97 consecutive patients with AP were prospectively analysed. Severe AP was defined according to the Atlanta Criteria. BMI was also calculated. To measure plasma Leptin, Nesfatin-<em>1</em> and Ghrelin concentrations, the blood samples were obtained from patients within 24 hours of admission.

RESULTS

Out of 97 patients, 92(70 females, 22 males) were considered eligible for analysis. Of the 92 patients, 30 patients (32.6%) were assessed as severe pancreatitis. BMI and leptin levels were significantly higher in patients with severe pancreatitis. The pooled sensitivity and specificity of BMI as a predictor for the development of pancreatic necrosis were 0.90(95%CI = 0.56-0.99) and 0.70(95%CI = 0.58-0.79), respectively; with an overall area under curve value of 0.78.The pooled sensitivity and specificity of leptin levels as a predictor for development of pancreatic necrosis were <em>1</em>(95%CI = 0.69-<em>1</em>) and 0.73(95%CI = 0.62-0.82),respectively; with an overall area under curve value of 0.82.Nesfatin-<em>1</em> and ghrelin levels showed no significant difference in patients with mild pancreatitis (6.97 ± 0.84 ng/ml and 2.3(<em>1</em>.0-9.9);respectively) and severe pancreatitis (6.74 ± 0.65 ng/ml and 2.0(<em>1</em>.9-9.9); respectively) (p = 0.<em>1</em>923 and 0.853<em>1</em>;respectively).

CONCLUSIONS

BMI and plasma leptin levels both were correlated with the severity of pancreatitis. Leptin levels showed better area under the curve, sensitivity and specificity values compared to BMI in prediction of pancreatic necrosis.Nesfatin-<em>1</em> and ghrelin levels were not found to be predictors of the severity of disease.

The phenotypic transformation from differentiated to dedifferentiated vascular smooth muscle cells (VSMCs) plays a crucial role in VSMC proliferation and vascular remodeling in many cardiovascular diseases including hypertension. <em>Nesfatin</em>-<em>1</em>, a multifunctional adipocytokine, is critically involved in the regulation of blood pressure. However, it is still largely unexplored whether <em>nesfatin</em>-<em>1</em> is a potential candidate in VSMC phenotypic switch and proliferation in hypertension. Experiments were carried out in Wistar-Kyoto rats (WKY), spontaneously hypertensive rats (SHR), human VSMCs and primary rat aortic VSMCs. We showed that the expression of <em>nesfatin</em>-<em>1</em> was upregulated in media layer of the aorta in SHR and SHR-derived VSMCs. <em>Nesfatin</em>-<em>1</em> promoted VSMC phenotypic transformation, accelerated cell cycle progression and proliferation. Knockdown of <em>nesfatin</em>-<em>1</em> inhibited the VSMC phenotype switch from a contractile to a synthetic state, attenuated cell cycle progression and retarded VSMC proliferation in SHR-derived VSMCs. Moreover, <em>nesfatin</em>-<em>1</em>-activated PI3K/Akt/mTOR signaling was abolished by JAK/STAT inhibitor WP<em>1</em>066, and the increased phosphorylation levels of JAK2/STAT3 in response to <em>nesfatin</em>-<em>1</em> were suppressed by inhibition of PI3K/Akt/mTOR in VSMCs. Pharmacological blockade of the forming feedback loop between PI3K/Akt/mTOR and JAK2/STAT3 prevented the proliferation of <em>nesfatin</em>-<em>1</em>-incubated VSMCs and primary VSMCs from SHR. Chronic intraperitoneal injection of <em>nesfatin</em>-<em>1</em> caused severe hypertension and cardiovascular remodeling in normal rats. In contrast, silencing of <em>nesfatin</em>-<em>1</em> gene ameliorated hypertension, phenotype switching, and vascular remodeling in the aorta of SHR. Therefore, our data identified <em>nesfatin</em>-<em>1</em> as a key modulator in hypertension and vascular remodeling by facilitating VSMC phenotypic switching and proliferation.

Brainstem structures such as the nucleus of the solitary tract (NTS) and the dorsal motor nucleus of the vagus nerve (DMNX) are essential for the digestive function of the stomach. A large number of neurotransmitters including glutamate and gamma-aminobutyric acid (GABA) are involved in the central control of gastric functions. However, the neuropeptidergic systems implicated in this process remain undetermined. <em>Nesfatin</em>-<em>1</em> was recently identified as a neuropeptide cleaved from the N-terminal part of NEFA/nucleobindin 2 precursor (NUCB2). Central administration of this neuropeptide inhibits food consumption and gastroduodenal motility in rodents. Interestingly, the NTS and the DMNX contain a dense population of NUCB2/<em>nesfatin</em>-<em>1</em> cell bodies. These observations led us to investigate the possible involvement of NUCB2/<em>nesfatin</em>-<em>1</em> neurons in the brainstem neuronal pathways that modulate gastric functions. We observed an activation of NTS NUCB2/<em>nesfatin</em>ergic neurons after gastric distention in rats. In addition, we found that several NTS NUCB2/<em>nesfatin</em>ergic neurons were GABAergic. Finally, when fluorogold was injected at the stomach level, many retrogradely labeled neurons were observed in the DMNX which were also positive for NUCB2/<em>nesfatin</em>-<em>1</em>. Taken together, these observations suggest for the first time that NUCB2/<em>nesfatin</em>-<em>1</em> neurons of the NTS are sensitive to gastric distension and then may contribute to the satiety signal.

The anorectic neuropeptide <em>nesfatin</em>-<em>1</em> has recently been characterized as a potential mood regulator, but the accurate effect of <em>nesfatin</em>-<em>1</em> on anxiety and learning and memory behavior and the possible mechanisms remains unknown. In the present study, to test the hypothesis that <em>nesfatin</em>-<em>1</em> might affect the anxiety-like and learning and memory behaviors in rats via ERK/CREB/BDNF pathway, <em>nesfatin</em>-<em>1</em> was administered intraperitoneally to rats with the doses (<em>1</em>0, 20, 40μg/kg), and the behavioral performance was tested using the open field task, the Morris water maze (MWM), and the Y maze. Moreover, the protein expression of brain-derived neurotrophic factor (BDNF), total and phosphorylated-ERK in the hippocampus and the prefrontal cortex (PFC) were evaluated. The results showed that chronic administration of <em>nesfatin</em>-<em>1</em> could decrease the moving distance, the duration in the center, and the frequencies of rearing and grooming in the open field task, decrease the moving distance, frequency, and the preference index of new arm in the Y maze, although there was no significant difference of the performance in the MWM task among groups. Furthermore, 3 weeks' consecutive administration of <em>nesfatin</em>-<em>1</em> resulted in the decrease of protein expression of BDNF and phosphorylated-ERK in the hippocampus and the PFC. These results provided evidence that exogenous <em>nesfatin</em>-<em>1</em> could decrease exploration and induce anxiety-like behavior in rats, the mechanism of which might be related to the reduced protein expression of BDNF and phosphorylated-ERK in the hippocampus and the PFC.

OBJECTIVE

To analyze the concentrations of <em>nesfatin</em>-<em>1</em> in maternal and cord serum, to evaluate the expression of <em>nesfatin</em>-<em>1</em> in subcutaneous adipose tissue (SAT) from pregnant women with gestational diabetes mellitus (GDM) and those with normal glucose tolerance (NGT).

METHODS

We studied a total of 50 GDM and 50 NGT subjects. The clinical features, serum <em>nesfatin</em>-<em>1</em>, homeostasis model assessment of insulin resistance (HOMA-IR), lipid profiles were measured at the third trimester of pregnancy. The expression of <em>nesfatin</em>-<em>1</em> in the SAT was determined by western blot.

RESULTS

Compared with the NGT group, the GDM group showed greater levels of serum <em>nesfatin</em>-<em>1</em>, adipocyte fatty acid binding protein (AFABP), and leptin; a greater level of cord blood <em>nesfatin</em>-<em>1</em>; and a higher level of expression in SAT (p < 0.05 or p < 0.0<em>1</em>). Fasting insulin (FI) (b = 0.3<em>1</em>7, p= 0.022) and body mass index (BMI) before delivery (b = 0.367, p=0.008) were independently associated with serum <em>nesfatin</em>-<em>1</em>. Nesfatin-<em>1</em> was the independent risk factor for GDM.

CONCLUSIONS

The GDM group had higher levels of maternal serum and cord blood <em>nesfatin</em>-<em>1</em>, and greater <em>nesfatin</em>-<em>1</em> expression in SAT. Nesfatin-<em>1</em> is closely related to obesity and IR in pregnancy.

<em>Nesfatin</em>-<em>1</em> is a recently discovered metabolic peptide hormone that decreases food intake after lateral, third, or fourth brain ventricle; cisterna magna; or paraventricular nucleus (PVN) injection in ad libitum fed rats. Additional micro-injection studies will improve the understanding of how <em>nesfatin</em>-<em>1</em> acts on the brain and define specific nuclei responsive to <em>nesfatin</em>-<em>1</em>, which will provide insight on its effects on food intake. We evaluated how <em>nesfatin</em>-<em>1</em> injection into the dorsal vagal complex (DVC) modulates food intake response in rats during the dark phase. Consistent with previous observations, <em>nesfatin</em>-<em>1</em>-injected rats significantly reduced cumulative food intake over a 5-h period in rats. Chronic administration of <em>nesfatin</em>-<em>1</em> into the DVC reduced body weight gain over a <em>1</em>0-day period. Because glucosensing neurons in the DVC are involved in glucoprivic feeding and homeostatic control of blood glucose, we examined the effect of <em>nesfatin</em>-<em>1</em> on the excitability of DVC glucosensing neurons. <em>Nesfatin</em>-<em>1</em> inhibited most of the glucose-inhibitory (GI) neurons and excited most of the glucose-excitatory (GE) neurons in the DVC. Current-clamp electrophysiology recordings from DVC glucosensing neurons in slice preparation showed that bath applied <em>nesfatin</em>-<em>1</em>(<em>1</em>0 nM) increased the firing frequency of GE neurons and inhibited the firing rate of GI-neurons. <em>Nesfatin</em>-<em>1</em> inhibited 88.9% (<em>1</em>6/<em>1</em>8) of gastric distension inhibitory (GD-INH) neurons and excited 76.2% (32/42) of gastric distension excitatory (GD-EXC) neurons. Thus, <em>nesfatin</em>-<em>1</em> may control food intake by modulating the excitability of glucosensing neurons in the DVC.

Osteoarthritis (OA) is a chronic degenerative joint disease, related to the overexpression of matrix metalloproteinases (MMPs), a disintegrin and metalloproteinase with thrombospondin motifs (ADAMTS), inflammation, and chondrocyte apoptosis. <em>Nesfatin</em>-<em>1</em> is an adipokine, which plays an important role in the development of OA, especially in obese people. In the present study, cartilage degradation and apoptosis observed in OA patients was evaluated. Furthermore, the anti-inflammatory and anti-apoptotic effects of <em>nesfatin</em>-<em>1</em>, and its underlying in vitro and in vivo mechanisms were investigated. The results showed that <em>nesfatin</em>-<em>1</em> increased significantly the expression of collagen type II alpha <em>1</em> chain (Col2a<em>1</em>), and reduced the expression of MMPs, ADAMTS5, cyclooxygenase (COX)-2, caspase-3, nitric oxide (NO), inducible nitric oxide synthase (iNOS), prostaglandin E2 (PGE2), interleukin (IL)-6, and chondrocyte apoptosis rate, which may be induced by IL-<em>1</em>β in rat chondrocytes. Furthermore, <em>nesfatin</em>-<em>1</em> treatment prevented cartilage degeneration in the rat OA model. It was found that <em>nesfatin</em>-<em>1</em> suppressed the IL-<em>1</em>β-induced activation of NF-κB, the mitogen-activated protein kinase (MAPK), and the Bax/Bcl-2 signal pathway in chondrocytes. These results suggest that in vivo <em>nesfatin</em>-<em>1</em> could play a protective role in the development of OA and can be potentially used for its treatment.

This study examines the levels of acylated and desacylated ghrelin, preptin, leptin, and <em>nesfatin</em>-<em>1</em> peptide changes related to the body mass index (BMI). The subjects were allocated to 5 groups depending on their BMIs as follows: Group I (BMI (<em>1</em>8.5 kg/m(2)); Group II (BMI <em>1</em>8.5-24.9 kg/m(2)); Group III (BMI 25-29.9 kg/m(2)); Group IV (BMI 30-39.9 kg/m(2)); Group V (BMI >40 kg/m(2)). Serum acylated and desacylated ghrelin, preptin, and leptin levels were measured by the enzyme-linked immunosorbent assay (ELISA) and <em>nesfatin</em>-<em>1</em> was measured by the enzyme immunoassay (EIA). Desacylated ghrelin levels showed a gradual and statistically significant drop from Group I to Group V, while preptin and leptin levels exhibited a gradual and significant increase from Group I to Group IV. Serum <em>nesfatin</em>-<em>1</em> levels gradually, but not significantly, increased from Group I to Group III and showed a significant decrease in Groups IV and V. In conclusion, leptin, preptin, and acylated ghrelin (AG) levels increased with higher BMI, whereas desacylated ghrelin (DAG) decreased and <em>nesfatin</em>-<em>1</em> showed no clear relationship to BMI.