Nucleobindin 2 (NUCB2) is a precursor of <em>nesfatin</em>-<em>1</em>, a hypothalamic anorectic neuropeptide. The association between variants of the NUCB2 gene and adiposity was examined. <em>1</em>42 severely obese Chinese children in Singapore, and 384 normal weight Chinese children from a longitudinal cohort from Da Qing, China, were studied. NUCB2 was screened using PCR and direct sequencing in 29 severely obese children and 24 non-obese children, then screened for a variant c.<em>1</em>0<em>1</em>2C>G (Q338E, or rs75708<em>1</em>) in the rest of the cohort using TaqMan probe. Five variants, including c.<em>1</em>0<em>1</em>2C>G (Q338E) were found. Genotyping for c.<em>1</em>0<em>1</em>2C>G found that the GG genotype was significantly less frequent in the obese group; odds ratio for obese subjects carrying the CC and CG genotypes was 2.29 (95% CI <em>1</em>.<em>1</em>7-4.49) in the dominant model, CC genotype 2.86 (95% CI <em>1</em>.4<em>1</em>-5.8<em>1</em>) in the additive model, and C allele <em>1</em>.57 (95% CI <em>1</em>.<em>1</em>7-2.<em>1</em>). The findings were replicated in an independent cohort of 372 obese and 390 normal weight Chinese children, where the odds ratio of obese subjects with CC and CG genotypes was <em>1</em>.69 (95% CI <em>1</em>.<em>1</em>2-2.55). Within the Da Qing cohort, subjects with the GG genotype had significantly lower BMI and percentage ideal weight for height (WFH) at 5 and 8years of age. Subjects with lower birth weights also had more pronounced difference in WFH and BMI at 5 and <em>1</em>0years of age between GG subjects versus CC/CG subjects. We postulate that GG genotype is protective against excessive weight gain, and factors which predispose to excessive weight gain such as higher birth weights may ameliorate the effect.

Nucleobindin-2 is a 420 amino acid EF-hand Ca²⁺ binding protein that can be further processed to generate an 82 amino terminal peptide termed <em>Nesfatin</em>-<em>1</em>. To examine the function of secreted Nucleobindin-2 in adipocyte differentiation, cultured 3T3-L<em>1</em> cells were incubated with either 0 or <em>1</em>00 nM of GST, GST-Nucleobindin-2, prior to and during the initiation of adipocyte differentiation. Nucleobindin-2 treatment decreased neutral lipid accumulation (Oil-Red O staining) and expression of several marker genes for adipocyte differentiation (PPARγ, aP2, and adipsin). When Nucleobindin- 2 was constitutively secreted into cultured medium, cAMP content and insulin stimulated CREB phosphorylation were significantly reduced. On the other hand, intracellularly overexpressed Nucleobindin-2 failed to affect cAMP content and CREB phosphorylation. Taken together, these data indicate that secreted Nucleobindin-2 is a suppressor of adipocyte differentiation through inhibition of cAMP production and insulin signal.

<em>Nesfatin</em>-<em>1</em>/NucB2, an anorexigenic molecule, is expressed mainly in the hypothalamus, particularly in the supraoptic nucleus (SON) and the paraventricular nucleus (PVN). <em>Nesfatin</em>-<em>1</em>/NucB2 is also expressed in the subfornical organ (SFO). Because the SON and PVN are involved in body fluid regulation, <em>nesfatin</em>-<em>1</em>/NucB2 may be involved in dehydration-induced anorexia. To clarify the effects of endogenous <em>nesfatin</em>-<em>1</em>/NucB2, we studied changes in <em>nesfatin</em>-<em>1</em>/NucB2 mRNA levels in the SFO, SON, and PVN in adult male Wistar rats after exposure to osmotic stimuli by using in situ hybridization histochemistry. Significant increases in <em>nesfatin</em>-<em>1</em>/NucB2 mRNA levels, ∼2- to 3-fold compared with control, were observed in the SFO, SON, and PVN following water deprivation for 48 h, consumption of 2% NaCl hypertonic saline in drinking water for 5 days, and polyethylene glycol-induced hypovolemia. In addition, <em>nesfatin</em>-<em>1</em>/NucB2 expression was increased in response to water deprivation in a time-dependent manner. These changes in <em>nesfatin</em>-<em>1</em>/NucB2 mRNA expression were positively correlated with plasma sodium concentration, plasma osmolality, and total protein levels in all of the examined nuclei. Immunohistochemistry for <em>nesfatin</em>-<em>1</em>/NucB2 revealed that <em>nesfatin</em>-<em>1</em>/NucB2 protein levels were also increased after 48 h of dehydration and attenuated by 24 h of rehydration. Moreover, intracerebroventricular administration of <em>nesfatin</em>-<em>1</em>/NucB2-neutralizing antibody after 48 h of water deprivation resulted in a significant increase in food intake compared with administration of vehicle alone. These results suggested that <em>nesfatin</em>-<em>1</em>/NucB2 is a crucial peptide in dehydration-induced anorexia.

OBJECTIVE

Bariatric surgery has been reported to be very effective in the remission of type 2 diabetes mellitus (T2DM). However, the mechanism is still under debate. <em>Nesfatin</em>-<em>1</em>, a recently discovered anorexigenic neuropeptide, was reported to be very important in glucose metabolism and regulating food intake. In this study, the effects of bariatric surgery on the expression and regulation of <em>nesfatin</em>-<em>1</em> were discussed.

METHODS

T2DM was induced in SD rats by a diet high in sugar and fat plus a low dose of streptozotocin (STZ) (25 mg/kg) injection. Bariatric surgeries, including Roux-En-Y Gastric Bypass (RYGB) and sleeve gastrectomy (SG), were performed on these rats. Two months after the surgery, the plasma <em>nesfatin</em>-<em>1</em> level and the expression of <em>nesfatin</em>-<em>1</em> in different organs of the rats were tested. Next, in vivo administration of <em>nesfatin</em>-<em>1</em> after surgery was performed to investigate the role of <em>nesfatin</em>-<em>1</em> in bariatric surgery.

RESULTS

Both RYGB and SG could reduce the weight of the rats. However, only RYGB had significant effects on the blood glucose level. Neither surgeries seemed to affect the blood concentration of insulin. However, RYGB significantly improved insulin sensitivity. Expression of <em>nesfatin</em>-<em>1</em> in the plasma and relative organs decreased in T2DM rats and rose again after RYGB; however, this pattern did not occur in SG. Injection of <em>nesfatin</em>-<em>1</em> after SG significantly improved insulin resistance and reduced blood glucose levels.

CONCLUSIONS

<em>Nesfatin</em>-<em>1</em> may improve insulin sensitivity in T2DM rats and thus plays a very important role in the remission of T2DM after RYGB. This neuropeptide could be a new target for directing future improvements in the bariatric surgical process.

<em>Nesfatin</em>-<em>1</em> is a polypeptide derived from the posttranslational processing of the N-terminal fragment of nucleobindin 2 (NUCB2), that was originally identified as an anorexigenic hypothalamic neuropeptide. A number of reports have recently shown that NUCB2/<em>nesfatin</em>-<em>1</em> is widely expressed in various peripheral tissues, including those of the gastrointestinal tract where it may participate in various pathophysiological processes. One of its roles may be regulation of energy homeostasis. As a result, <em>nesfatin</em>-<em>1</em> may be a novel target for exploring the underlying mechanisms and the treatment of metabolic syndromes.

In this article, we review on the current concepts about <em>Nesfatin</em>-<em>1</em> as a new anti-obesity treatment and evaluate the existing issues in the context of this knowledge and the available literature. The intent is to enable clinicians to know <em>Nesfatin</em>-<em>1</em> as a new anti-obesity treatment and make rational decisions based on this perspective as possible clinical application. Future research should seek to clarify whether <em>Nesfatin</em>-<em>1</em> would be beneficial in the management of obesity.

OBJECTIVE

<em>Nesfatin</em>-<em>1</em> (NF-<em>1</em>), an anorexic nucleobindin-2 (NUCB2)-derived hypothalamic peptide, acts as a peripheral cardiac modulator and it can induce negative inotropic effects. However, the mechanisms underlying these effects in cardiomyocytes remain unclear.

METHODS

Using patch clamp, protein kinase assays, and western blot analysis, we studied the effect of NF-<em>1</em> on L-type Ca2+ currents (ICa,L) and to explore the regulatory mechanisms of this effect in adult ventricular myocytes.

RESULTS

NF-<em>1</em> reversibly decreased ICa,L in a dose-dependent manner. This effect was mediated by melanocortin 4 receptor (MC4-R) and was associated with a hyperpolarizing shift in the voltage-dependence of inactivation. Dialysis of cells with GDP-β-S or anti-Gβ antibody as well as pertussis toxin pretreatment abolished the inhibitory effects of NF-<em>1</em> on ICa,L. Protein kinase C (PKC) antagonists abolished NF-<em>1</em>-induced responses, whereas inhibition of PKA activity or intracellular application of the fast Ca2+-chelator BAPTA elicited no such effects. Application of NF-<em>1</em> increased membrane abundance of PKC theta isoform (PKCθ), and PKCθ inhibition abolished the decrease in ICa,L induced by NF-<em>1</em>.

CONCLUSIONS

These data suggest that NF-<em>1</em> suppresses L-type Ca2+ channels via the MC4-R that couples sequentially to the βγ subunits of Gi/o-protein and the novel PKCθ isoform in adult ventricular myocytes.

Body weight is related to fat mass, which is associated with obesity. Our study explored the effect of fat-<em>1</em> gene on body weight in fat-<em>1</em> transgenic mice. In present study, we observed that the weight/length ratio of fat-<em>1</em> transgenic mice was lower than that of wild-type mice. The serum levels of triglycerides (TG), cholesterol (CT), high-density lipoprotein cholesterol (HDL-c), low-density lipoprotein cholesterol (LDL-c) and blood glucose (BG) in fat-<em>1</em> transgenic mice were all decreased. The weights of peri-bowels fat, perirenal fat and peri-testicular fat in fat-<em>1</em> transgenic mice were reduced. We hypothesized that increase of n-3 PUFAs might alter the expression of hypothalamic neuropeptide genes and lead to loss of body weight in fat-<em>1</em> transgenic mice. Therefore, we measured mRNA levels of appetite neuropeptides, Neuropeptide Y (NPY), Agouti-related peptides (AgRP), Proopiomelanocortin (POMC), Cocaine and amphetamine regulated transcript (CART), ghrelin and <em>nesfatin</em>-<em>1</em> in hypothalamus by real-time PCR. Compared with wild-type mice, the mRNA levels of CART, POMC and ghrelin were higher, while the mRNA levels of NPY, AgRP and <em>nesfatin</em>-<em>1</em> were lower in fat-<em>1</em> transgenic mice. The results indicate that fat-<em>1</em> gene or n-3 PUFAs participates in regulation of body weight, and the mechanism of this phenomenon involves the expression of appetite neuropeptides and lipoproteins in fat-<em>1</em> transgenic mice.

BACKGROUND

Overweight and diseases connected with it are an increasing problem in children and adolescents. Thyroid disease leads to a change of weight - in hyperthyroidism body mass is reduced whereas in hypothyroidism it is increased. It is emphasized that changes in hormones such as peptide levels are in close relationship with the regulation of body mass. <em>Nesfatin</em>-<em>1</em> is a recently described anorexigenic peptide produced by the brain. <em>Nesfatin</em>-<em>1</em> also reduces body weight gain, suggesting a role as a new modulator of energy balance. Excess <em>nesfatin</em> in the brain leads to a loss of appetite, less frequent hunger, a `sense of fullness´, and a drop in body fat and weight. A lack of <em>nesfatin</em>-<em>1</em> in the brain leads to an increase of appetite, more frequent episodes of hunger, an increase of body fat and weight, and the inability to `feel full´. Aim of the study was to evaluate <em>nesfatin</em>-<em>1</em> levels in young patients with untreated Graves´ disease, subclinical Hashimoto´ thyroiditis, and in healthy children.

METHODS

The study group formed 78 patients of the Outpatient Endocrinology Clinic of Pediatrics, Endocrinology, Diabetology with Cardiology Division. In all the patients, nesfatin level was analyzed by the ELISA´s method.

RESULTS

In the group with hyperthyroidism in Graves´ disease lower levels of <em>nesfatin</em>-<em>1</em> were found compared to the group of healthy children (<em>1</em>9.37 vs 32.96 ng/ml; p<0.02); after appropriate treatment in that group the levels of <em>nesfatin</em>-<em>1</em> were higher compared to the group with hyperthyroidism, but lower compared to the group of healthy children (20.35 vs 32.96 ng/ml; NS). On the other hand, <em>nesfatin</em>-<em>1</em> levels were lower in children with untreated subclinical hypothyroidism in Hashimoto´s thyroiditis compared to the group of healthy children (<em>1</em>7.2 vs32.96 ng/ml; p<0.002). After treatment of L-thyroxine lower levels of <em>nesfatin</em>-<em>1</em> were found compared to the control group (<em>1</em>4.5 vs 32.96 ng/ml; NS). No relationship between <em>nesfatin</em>-<em>1</em> and thyroid hormones was observed.

CONCLUSIONS

It might be that disturbances in thyroid hormones in thyroid diseases do not have an essential effect on changes of <em>nesfatin</em>-<em>1</em> - an appetite-controlling hormone/peptide. Secondly, <em>nesfatin</em>-<em>1</em> levels were lower in children with untreated autoimmune thyroid diseases, however, the mechanism is also unknown.

Originally identified in the hypothalamus as a satiety factor, recent studies provide evidence that nefatin-<em>1</em>/NUCB2 is a gutbrain peptide with a broader array of actions. Detection of abundant <em>nesfatin</em>-<em>1</em>/NUCB2 in gastric X/A like endocrine cells, which also produce the orexigenic hormone ghrelin, indicates that gastric mucosa may be one of the predominant sources of <em>nesfatin</em>-<em>1</em>/NUCB2. Functional studies have revealed significant effects of nefatin-<em>1</em> on inhibition of feeding behavior and on glucose homeostasis. These metabolic functions make <em>nesfatin</em>-<em>1</em>/NUCB2 a novel candidate for treatment of obesity and diabetes. However, deficiencies in our understanding of <em>nesfatin</em>-<em>1</em>/NUCB2 receptor pose a significant hurdle for therapies that target its action. Defining novel pathways to alter the production of <em>nesfatin</em>-<em>1</em>/NUCB2 would shift therapeutic focus to gastric targets. A necessary precondition is improved understanding of the mechanisms by which <em>nesfatin</em>-<em>1</em>/NUCB2 is synthesized and secreted by gastric X/A like cells. Recent studies provide evidence that mTOR is a critical regulatory molecule in these endocrine cells and that its activity is linked to the production of ghrelin and <em>nesfatin</em>- <em>1</em>/NUCB2. These findings suggest that gastric mTOR is involved in the regulation of food intake and overall energy metabolism through modulation of ghrelin and <em>nesfatin</em>-<em>1</em>/NUCB2. In this review, we first summarize current advances in the relationship between organism energy status and <em>nesfatin</em>-<em>1</em>/NUCB2 levels, and then discuss the novel finding on mTOR as the gastric fuel sensor and its role in the regulation of <em>nesfatin</em>-<em>1</em>/NUCB2 expression.

Osteoarthritis (OA) is a common joint disease for which a safe and reliable treatment has yet to be developed. Here, we demonstrated the potential benefit of treatment with paeonol, a derivative of <i>Paeonia suffruticosa</i>, in the treatment and prevention of OA. Chondrogenic cell line ATDC5 cells were cultured with IL-<em>1</em>β and the effects of paeonol were assessed through qRT-PCR, western blot analysis, MTT, ELISA, and NF-κB luciferase reporter gene assay. Our findings demonstrate a novel ability of paeonol to inhibit numerous factors of OA, including expressions of IL-6, TNF-α, NOX2, PTGS2, NUCB2/<em>nesfatin</em>-<em>1</em>, ICAM-<em>1</em>, VCAM-<em>1</em>, MMP-3/<em>1</em>3, degradation of type II collagen, and NF-κB activation through the rescue of IκBα. Additionally, we assessed the effects of paeonol on cell viability to confirm its safety. These findings implicate a valuable potential role of paeonol in the treatment and prevention of OA.

Ghrelin is an orexigenic peptide hormone produced mainly by a distinct group of dispersed endocrine cells located within the gastric oxyntic mucosa. Besides secreted gene products derived from the preproghrelin gene, which include acyl-ghrelin, desacyl-ghrelin and obestatin, ghrelin cells also synthesize the secreted protein <em>nesfatin</em>-<em>1</em>. The main goal of the current study was to identify other proteins secreted from ghrelin cells. An initial gene chip screen using mRNAs derived from highly enriched pools of mouse gastric ghrelin cells demonstrated high levels of serum retinol-binding protein (RBP4) and transthyretin (TTR), both of which are known to circulate in the bloodstream bound to each other. This high expression was confirmed by quantitative RT-PCR using as template mRNA derived from the enriched gastric ghrelin cell pools and from two ghrelin-producing cell lines (SG-<em>1</em> and PG-<em>1</em>). RBP4 protein also was shown to be secreted into the culture medium of ghrelin cell lines. Neither acute nor chronic caloric restriction had a significant effect on RBP4 mRNA levels within stomachs of C57BL/6J mice, although both manipulations significantly decreased stomach TTR mRNA levels. In vitro studies using PG-<em>1</em> cells showed no effect on RBP4 release of octanoic acid, epinephrine or norepinephrine, all of which are known to act directly on ghrelin cells to stimulate ghrelin secretion. These data provide new insights into ghrelin cell physiology, and given the known functions of RBP4 and TTR, support an emerging role for the ghrelin cell in blood glucose handling and metabolism.

BACKGROUND Psoriasis is an autoimmune, inflammatory, and chronic disease. Recent studies have evaluated serum endocan and <em>nesfatin</em>-<em>1</em> levels in patients with inflammatory disorders. The neutrophil-to-lymphocyte ratio (NLR) is an inflammatory marker currently used in many diseases. The aim of the present study was to evaluate NLR, serum endocan, and <em>nesfatin</em>-<em>1</em> levels in psoriasis vulgaris before and after narrow-band ultraviolet B (NB-UVB) phototherapy treatment and compared to healthy controls. MATERIAL AND METHODS This study was conducted on a total of 88 cases, 39 of which had psoriasis vulgaris and 49 were healthy volunteers. Thirty-nine psoriasis vulgaris patients underwent NB-UVB phototherapy treatment for 3 months. NLR, serum endocan, and <em>nesfatin</em>-<em>1</em> levels were measured in all psoriasis patients before and after NB-UVB phototherapy and in the control group. RESULTS Compared with the control group, neutrophil count and NLR were significantly higher (p<0.00<em>1</em>) in psoriasis patients before NB-UVB phototherapy. Serum endocan levels were significantly correlated with disease activity before treatment. There was no significant difference in NLR, serum endocan, and <em>nesfatin</em>-<em>1</em> levels in psoriasis patients before and after NB-UVB phototherapy (p>0.05). CONCLUSIONS The current study shows that NLR was higher in psoriasis vulgaris patients when compared with the control group, whereas serum endocan and <em>nesfatin</em>-<em>1</em> levels were not significantly different. In addition, NB-UVB phototherapy did not affect NLR, serum endocan, or <em>nesfatin</em>-<em>1</em> levels. Further larger-scale studies are required on this subject.

<em>Nesfatin</em>-<em>1</em>/NUCB2 is a neuropeptide that plays important roles in regulating food intake and energy homeostasis. The distribution of <em>nesfatin</em>-<em>1</em>/NUCB2 protein and mRNA has not been investigated in the canine digestive system. The present study was conducted to evaluate the expression of <em>nesfatin</em>-<em>1</em>/NUCB2 protein and NUCB2 mRNA in the canine digestive organs (esophagus, stomach, duodenum, jejunum, ileum, cecum, colon, rectum, liver and pancreas). The tissues of the digestive system were collected from dogs at different developmental stages (infantile, juvenile, pubertal and adult). <em>Nesfatin</em>-<em>1</em>/NUCB2 protein localization in the organs of adult dogs was detected by immunohistochemistry. The expression of NUCB2 mRNA at the four developmental stages was analyzed by real-time fluorescence quantitative PCR (qRT-PCR). <em>Nesfatin</em>-<em>1</em>/NUCB2 protein was distributed in the fundic gland region of the stomach, and the islet area and exocrine portions of the pancreas. However, NUCB2 mRNA was found in all digestive organs, although the expression levels in the pancreas and stomach were higher than those in liver, duodenum and other digestive tract tissues (P<0.05) at the four different developmental stages of the dogs. In this study, <em>nesfatin</em>-<em>1</em>/NUCB2 was found to be present at high levels in the stomach and pancreas at both the protein and mRNA levels; however, NUCB2 expression was found at lower levels in all of the digestive organs. These findings provide the basis of further investigations to elucidate the functions of nefatin-<em>1</em> in the canine digestive system.

BACKGROUND

Despite of its significant therapeutic effects on obesity and metabolic diseases, Roux-en-Y gastric bypass (RYGB) has limited clinical application because of considerable impacts on the gastrointestinal structure and postoperative complications. This study aims to develop a simplified surgical approach with less damage and complication but efficient metabolic benefit.

METHODS

The effects of Esophagus-Duodenum gastric bypass (EDGB) on body weight, food intake, glucose and lipid metabolism were compared to RYGB in mice.

RESULTS

EDGB is simple, has higher survival rate and less complication. Relative to RYGB, EDGB demonstrated modest body weight control, identical improvement of glucose and lipid metabolism in obese mice. Blood glucose increased significantly <em>1</em>5 and 30min after oral glucose administration, then markedly decreased in both EDGB and RYGB groups relative to the sham surgery, indicating a quicker absorption of oral glucose and improvement in glucose uptake by insulin targeted tissues. Insulin sensitivity was identically improved. EDGB significantly decreased plasma and hepatic triglyceride levels, while increased browning in visceral and subcutaneous white adipose tissue to the extent identical to RYGB. Levels of ghrelin and <em>nesfatin</em>-<em>1</em> increased significantly after EDGB and RYGB.

CONCLUSIONS

EDGB is a valuable model to study the metabolic benefit of bariatric surgery in mice.

<em>Nesfatin</em>-<em>1</em> is secreted, meal-responsive anorexigenic peptide encoded in the precursor nucleobindin-2 [NUCB2]. Circulating <em>nesfatin</em>-<em>1</em> increases post-prandially, but the dietary components that modulate NUCB2/<em>nesfatin</em>-<em>1</em> remain unknown. We hypothesized that carbohydrate, fat and protein differentially regulate tissue specific expression of <em>nesfatin</em>-<em>1</em>. NUCB2, prohormone convertases and <em>nesfatin</em>-<em>1</em> were detected in mouse stomach ghrelinoma [MGN3-<em>1</em>] cells. NUCB2 mRNA and protein were also detected in mouse liver, and small and large intestines. MGN3-<em>1</em> cells were treated with glucose, fatty acids or amino acids. Male C57BL/6 mice were chronically fed high fat, high carbohydrate and high protein diets for <em>1</em>7 weeks. Quantitative PCR and <em>nesfatin</em>-<em>1</em> assays were used to determine <em>nesfatin</em>-<em>1</em> at mRNA and protein levels. Glucose stimulated NUCB2 mRNA expression in MGN3-<em>1</em> cells. L-Tryptophan also increased NUCB2 mRNA expression and ghrelin mRNA expression, and <em>nesfatin</em>-<em>1</em> secretion. Oleic acid inhibited NUCB2 mRNA expression, while ghrelin mRNA expression and secretion was enhanced. NUCB2 mRNA expression was significantly lower in the liver of mice fed a high protein diet compared to mice fed other diets. Chronic intake of high fat diet caused a significant reduction in NUCB2 mRNA in the stomach, while high protein and high fat diet caused similar suppression of NUCB2 mRNA in the large intestine. No differences in serum <em>nesfatin</em>-<em>1</em> levels were found in mice at 7 a.m, at the commencement of the light phase. High carbohydrate diet fed mice showed significantly elevated <em>nesfatin</em>-<em>1</em> levels at <em>1</em> p.m. Serum <em>nesfatin</em>-<em>1</em> was significantly lower in mice fed high fat, protein or carbohydrate compared to the controls at 7 p.m, just prior to the dark phase. Mice that received a bolus of high fat had significantly elevated <em>nesfatin</em>-<em>1</em>/NUCB2 at all time points tested post-gavage, compared to control mice and mice fed other diets. Our results for the first time indicate that <em>nesfatin</em>-<em>1</em> is modulated by nutrients.

The cachexia occurs frequently in lung cancer patients. Among appetite regulatory peptides, alteration of expressions of leptin and ghrelin is demonstrated in cachectic cancer patients, but <em>nesfatin</em>-<em>1</em> has not been yet studied in cancer. We investigated serum <em>nesfatin</em>-<em>1</em> level in advanced lung cancer patients. Forty-one lung cancer patients and 24 healthy subjects were included to the study. <em>Nesfatin</em>-<em>1</em> serum levels were analyzed by ELISA kit. Serum <em>nesfatin</em>-<em>1</em> levels were lower in lung cancer patients than in healthy subjects (0.52±0.<em>1</em>9ng/ml vs 0.75±0.23ng/ml; p<0.00<em>1</em>). In lung cancer patients with weight loss, <em>nesfatin</em>-<em>1</em> levels were decreased compared to the patients without weight loss (0.44±0.<em>1</em>6ng/ml vs 0.63±0.<em>1</em>8ng/ml; p<0.00<em>1</em>). Whereas, there were no any difference between patients without weight loss and control subjects (0.63±0.<em>1</em>8ng/ml vs 0.75±0.23ng/ml; p:0.<em>1</em>29) or between SCLC and NSCLC patients (0.53±0.<em>1</em>8ng/ml vs 0.52±0.20ng/ml; p:0.458). No significant correlation was found between serum <em>nesfatin</em>-<em>1</em> values and BMI. In conclusion, loss of fat mass may decrease serum <em>nesfatin</em>-<em>1</em> level in lung cancer patients with weight loss. The future studies which explore biological significance of low serum <em>nesfatin</em>-<em>1</em> level in cancer are needed.

BACKGROUND

Hashimoto's thyroiditis is associated with serious alterations in serum lipids and glucose homeostasis. The aims of the current study were to evaluate the effect of powdered Nigella sativa on serum lipids, glucose homeostasis and anthropometric variables in patients with Hashimoto's thyroiditis.

METHODS

Forty patients with Hashimoto's thyroiditis, aged between 22 and 50 years old, participated in the trial and were randomly allocated into two groups of intervention and control receiving powdered Nigella sativa or placebo daily for 8 weeks. Serum lipids, glucose homeostasis, and anthropometric variables were evaluated at baseline and after intervention.

RESULTS

Treatment with Nigella sativa significantly reduced body weight and body mass index (BMI). Serum concentrations of low density lipoprotein cholesterol (LDL) and triglyceride (TG) also decreased in Nigella sativa-treated group after 8 weeks; while serum high density lipoprotein cholesterol (HDL) significantly increased after treatment with Nigella sativa (P < 0.05). None of these changes had been observed in placebo treated group. Serum <em>Nesfatin</em>-<em>1</em> concentrations was in inverse relationship with serum triglyceride (TG) (r = - 0.3<em>1</em>, P = 0.04).

CONCLUSIONS

Giving attention to the potent beneficial effects of powdered black cumin seeds in improving serum lipid profile and anthropometric features in patients with Hashimoto's thyroiditis, this medicinal plant could be considered as a beneficial herbal supplement alongside with the disease- specific medications including Levothyroxine in management of Hashimoto's thyroiditis- related metabolic abnormalities.

BACKGROUND

Iranian registry of clinical trials (registration number IRCT20<em>1</em>40908<em>1</em>9082N2 - Registered 20<em>1</em>4-09-29).

<em>Nesfatin</em>-<em>1</em>, a recently discovered peptide, is involved in important functions such as food intake regulation and energy homeostasis. Previous studies have demonstrated that it has protective effects following myocardial injury and also protects dopaminergic cells against neurotoxicity with the anti-inflammatory and anti-apoptotic mechanisms. In this study, we aimed to assay the neuroprotective effects of <em>Nesfatin</em>-<em>1</em> after brain ischemia/reperfusion. Twenty-eight male Wistar rats were randomly selected and allocated in the form of four groups (sham, <em>Nesfatin</em>-<em>1</em>, ischemia, ischemia+<em>Nesfatin</em>-<em>1</em>). Ischemia was created by obstruction couple common carotid arteries in 20-min period. Saline as a vehicle and <em>Nesfatin</em>-<em>1</em> (20 μg/kg, intraperitoneally) were injected at the time of reperfusion. Spatial memory performances were evaluated by the Morris water maze. The level of protein expression was determined by immunohistochemical and immunofluorescence staining. <em>Nesfatin</em>-<em>1</em> significantly reduced caspase-3 (P < 0.0<em>1</em>) and microglial activation (P < 0.0<em>1</em>) and improved spatial memory impairments (P < 0.05) induced by brain ischemia. <em>Nesfatin</em>-<em>1</em> has significant neuroprotective effects and can be introduced as a therapeutic agent against cerebral ischemia-induced injuries.

Metabolic homeostasis is achieved by proper functioning of a complex endocrine milieu, comprising of signals arising from the brain and peripheral tissues. Our knowledge of the factors regulating such balance is rapidly evolving, as new signaling molecules are being characterized. Of central interest is <em>nesfatin</em>-<em>1</em>, owing to its multifunctional tissue specific actions regulating food intake, body weight, blood glucose and cardiac functions.

This review discusses the tissue/system wide distribution and actions of <em>nesfatin</em>-<em>1</em> in regulating metabolic and cardiovascular functions in healthy conditions and diseases.

Nesfatin-<em>1</em> is an 82 amino acid peptide encoded in the secreted precursor, nucleobinin-2 (NUCB2). It was first reported as a novel anorexigen and a fat reducing orphan ligand. Research to date has established <em>nesfatin</em>-<em>1</em> in the regulation of food intake via modulation of neuropeptides in the feeding centers of brain. Nesfatin-<em>1</em> expression was also reported to have a spectrum of peripheral tissue specific actions, which includes insulin secretion (pancreas), fat and glucose modulation (adipocyte), gastric motility and gastric acid secretion (stomach), hormone secretion and transit time (intestine) and mean arterial pressure and cardiac injury (heart). Abnormal levels of <em>nesfatin</em>-<em>1</em> in circulation and/or variations in tissue specific expression have been observed in obesity, diabetes and cardiovascular diseases.

The multifunctional biological actions of <em>nesfatin</em>-<em>1</em> propelled this peptide as a therapeutic target, and as a potential biomarker of diseases. However, a better and comprehensive understanding of tissue specific effects of <em>nesfatin</em>-<em>1</em> is critical prior to exploring its possible use in the detection and treatment of diseases.

An adipokine leptin plays a central role in the regulation of feeding and energy homeostasis via acting on the hypothalamus. However, its downstream neuronal mechanism is not thoroughly understood. The neurons expressing nucleobindin-2 (NUCB2)-derived <em>nesfatin</em>-<em>1</em> in the hypothalamic paraventricular nucleus (PVN) have been implicated in feeding and energy homeostasis. The present study aimed to explore the role of PVN NUCB2/<em>nesfatin</em>-<em>1</em> in the leptin action, by using adeno-associated virus (AAV) vectors encoding shRNA targeting NUCB2 (AAV-NUCB2-shRNA). Leptin directly interacted and increased cytosolic Ca(2+) in single neurons isolated from the PVN, predominantly in NUCB2/nesftin-<em>1</em>-immunoreactive neurons. Treatment with leptin in vivo and in vitro markedly increased NUCB2 mRNA expression in the PVN. Peripheral and central injections of leptin failed to significantly inhibit food intake in mice receiving AAV-NUCB2. These results indicate that PVN NUCB2/<em>nesfatin</em>-<em>1</em> is directly targeted by leptin, and mediates its anorexigenic effect.

Ingestion of food affects the secretion of hormones from specialized endocrine cells scattered within the intestinal mucosa. Upon release, these hormones mostly decrease food intake by signaling information to the brain. Although enteroendocrine cells in the small intestine were thought to represent the predominant gut-brain regulators of food intake, recent advances also established a major role for gastric hormones in these regulatory pathways. First and foremost, the gastric endocrine X/A-like cell was in the focus of many studies due to the production of ghrelin, which is until now the only known orexigenic hormone that is peripherally produced and centrally acting. Although X/A-cells were initially thought to only release one hormone that stimulates food intake, this view has changed with the identification of additional peptide products also derived from this cell, namely desacyl ghrelin, obestatin, and <em>nesfatin</em>-<em>1</em>. Desacyl ghrelin may play a counter-regulatory role to the food intake stimulatory effect of ghrelin. The same property was suggested for obestatin; however, this hypothesis could not be confirmed in numerous subsequent studies. Moreover, the description of the stomach as the major source of the novel anorexigenic hormone <em>nesfatin</em>-<em>1</em> derived from the NUCB2 gene further corroborated the assumption that the gastric X/A-like cell products are not only stimulant but also inhibitors of feeding, thereby acting as so far unique dual regulator of food intake located in a logistically important place where the gastrointestinal tract has initial contact with food.

<em>Nesfatin</em>-<em>1</em>, a newly discovered satiety molecule which reduces feeding behavior, has been recognized as a unique regulatory neuropeptide with its multiple roles, both central and peripheral. However, whether it had neuronal modulation effect on dopaminergic neurons is largely unknown. In the present study, using whole-cell patch clamp under current-clamp mode, we investigate the effects of <em>nesfatin</em>-<em>1</em> on the electrical activity of rat nigral dopaminergic neurons. <em>Nesfatin</em>-<em>1</em> could produce a resting membrane potential hyperpolarization on the majority of dopaminergic neurons tested. The spike frequency decreased by 23.<em>1</em>3 ± 5.93 and 43.20 ± 5.56 % in 5-nM and <em>1</em>0-nM <em>nesfatin</em>-<em>1</em> groups, respectively. These effects persisted in the presence of ionotropic glutamate and GABA receptor antagonists. Our study suggests that <em>nesfatin</em>-<em>1</em> postsynaptically inhibits the electrical activity of nigral dopaminergic neurons.

<b>Background:</b> Chronic stress is an important risk factor for depression. The <em>nesfatin</em>-<em>1</em> (NES<em>1</em>)-oxytocin (OT)-proopiomelanocortin (POMC) neural pathway, which is involved in the stress response, was recently shown to have an anorectic effect in the hypothalamus. Our previous study showed that Xiaoyaosan, a well-known antidepressant used in traditional Chinese medicine, effectively relieved appetite loss induced by chronic immobilization stress (CIS). However, whether Xiaoyaosan ameliorates depression-like behaviors and anorexia by regulating the NES<em>1</em>-OT-POMC neural pathway remains unclear. <b>Objective:</b> To investigate whether the antidepressant-like and anti-anorexia effects of Xiaoyaosan are related to the NES<em>1</em>-OT-POMC neural pathway in the hypothalamus. <b>Methods:</b> Rats were randomly divided into control, CIS, Xiaoyaosan treatment, and fluoxetine treatment groups. The rats in the CIS, Xiaoyaosan treatment, and fluoxetine treatment groups were subjected to CIS for 2<em>1</em> consecutive days, during which they were administered distilled water, a Xiaoyaosan decoction [3.854 g/(kg·d)] or fluoxetine [<em>1</em>.76 mg/(kg·d)], respectively, by gavage, and their body weights and food intake were monitored daily. The rats were subsequently subjected to the open field test and sucrose preference test. Then, the expression levels of corticosterone and NES<em>1</em> in the serum and the expression levels of NES<em>1</em>, OT, POMC, and melanocortin-4 receptor (MC4R) in the hypothalamus were determined by real-time fluorescence quantitative polymerase chain reaction, Western blot analysis, and immunochemistry. Furthermore, immunofluorescence double staining was used to determine whether related proteins in the hypothalamic NES<em>1</em>-OT-POMC neural pathway were co-expressed. <b>Results:</b> Compared to control rats, rats exposed to CIS exhibited gradually less food intake and lower body weights and significantly increased concentrations of NES<em>1</em> in the serum and paraventricular nucleus. Moreover, the expression levels of POMC, OT, and MC4R in the hypothalamus were significantly higher in the CIS group than those in the control group. However, these changes were reversed by pretreatment with Xiaoyaosan and fluoxetine. Specifically, the expression levels of members of the NES<em>1</em>-OT-POMC neural pathway were lower in the Xiaoyaosan-treated group than in the CIS group. <b>Conclusion:</b> Xiaoyaosan ameliorates CIS-induced depression-like behaviors and anorexia by regulating the NES<em>1</em>-OT-POMC neural pathway in the hypothalamus.