Alter Food Intake Research Articles

Clusterin levels in undernourished SH-SY5Y cells

Food-related disorders are increasingly common in developed societies, and the psychological component of these disorders has been gaining increasing attention. Both overnourishment with high-fat diets and perinatal undernourishment in mice have been linked to a higher motivation toward food, resulting in an alteration in food intake. Clusterin (CLU), a multifaced protein, is overexpressed in the nucleus accumbens (NAc) of over-fed rats, as well as in those that suffered chronic undernutrition. Moreover, an increase of this protein was observed in the plasma of obese patients with food addiction, suggesting the implication of CLU in this eating disorder. To characterize CLU’s cellular mechanisms, in vitro experiments of undernutrition were performed using dopaminergic SH-SY5Y cells. To mimic in vivo dietary conditions, cells were treated with different fetal bovine serum (FBS) concentrations, resulting in control (C group) diet (10% FBS), undernourishment (U group) diet (0.5% FBS), and undernourishment diet followed by restoration of control diet (UC group) (0.5 + 10% FBS). Undernourishment compromised cell viability and proliferation, and concomitantly increased CLU secretion as well as the cytosolic pool of the protein, while decreasing the mitochondrial level. The restoration of normal conditions tended to recover cell physiology, and the normal levels and distribution of CLU. This research study is a step forward toward the characterization of clusterin as a potential marker for food addiction and nutritional status.

213 Eating Patterns Associated with Sleep Duration, Insomnia, Daytime Sleepiness and Overall Sleep Quality at the US-Mexico Border

Abstract Introduction Previous studies have linked sleep to risk of diabetes and obesity, at least partially via alterations in food intake. Diabetes and obesity are common among Hispanics/Latinos, and studies are needed to better clarify the role of sleep for health among this group. Methods Data were collected from N=100 adults (age 18-60, 47% female) of Mexican descent in the city of Nogales, AZ (34% not born in the US). Surveys were presented in English or Spanish. Eating Patterns were assessed with the Three-Factor Eating Questionnaire (TFEQ), which resulted in a total score and subscales for “cognitive restraint,” “uncontrolled eating, “and “emotional eating.” Insomnia was assessed with the use of the Insomnia Severity Index (ISI), Sleepiness with the use of the Epworth Sleepiness Scale (ESS), Sleep quality with the use of the Pittsburgh Sleep Quality Index (PSQI), and weekday and weekend sleep duration with the use of the Sleep Timing Questionnaire (STQ). Covariates included: age, sex, Body Mass Index (BMI), education and immigrant status. Results When adjusted for age, sex and immigrant status (model-1), eating patterns were associated with greater insomnia (95%CI:[0.066,1.095];p=0.027), poorer sleep quality (95%CI:[0.170,1.456];p=0.014), sleepiness (95%CI[0.032,1.026];p=0.037), and weekend (but not weekday) sleep duration (95%CI:[-0.031,0.003];p=0.015). Further adjustment for education (model-2) revealed similar significant associations. Additional adjustment for BMI (model-3) revealed a change in daytime sleepiness, where no association was seen (95%CI:[-0.202,0.805];p=0.238). Regarding subscale scores, relationships were generally seen between sleep and both emotional eating and uncontrolled eating, but not cognitive restraint. However, after adjustment for BMI, there was a significant association between cognitive restraint and weekend sleep duration (95%CI:[-0.015,-0.001];p=0.030). Conclusion Greater insomnia, poorer sleep quality, increased daytime sleepiness and decreased weekend sleep duration were associated with eating patterns at the US Mexico border, particularly in terms of uncontrolled eating and emotional eating. This suggests possible mechanisms linking sleep and obesity in Hispanic/Latinos. Support (if any) Supported by T32HL007249, R01MD011600, R01DA051321

Thymic Stromal Lymphopoietin induces adipose loss through sebum hypersecretion

Abstract Obesity is a serious public health concern. Emerging studies indicate that the immune system can regulate weight and energy homeostasis. Here we show that the cytokine Thymic Stromal Lymphopoietin (TSLP) stimulates T cells to induce selective white adipose loss, which protects against obesity, improves glucose metabolism, and mitigates nonalcoholic steatohepatitis. Surprisingly, adipose loss was not caused by alterations in food intake, absorption, or energy expenditure. Rather, it was caused by the excessive loss of lipids through the skin as sebum, causing mice to develop a striking “greasy hair” phenotype. Inhibition of sebum secretion or depletion of T cells prevented TSLP-driven adipose loss. TSLP-driven white fat loss and sebum hypersecretion were both mediated via the direct activation of CD4+ or CD8+ T cells by TSLP in an antigen-independent manner, and these T cells could be seen infiltrating skin sebaceous glands. Furthermore, we also found that TSLP and T cells regulated sebum release and sebum-associated anti-microbial peptide expression in the skin at steady-state. In human skin, TSLP expression also correlated directly with sebum-associated gene expression. Together, our study establishes a paradigm in which adipose loss can be achieved by sebum hypersecretion and reveals a previously unknown role of adaptive immunity in skin barrier function through the regulation of sebum secretion.

Mitochondrial Uncoupling Reverses Sarcopenic Obesity while Enhancing Glucose Homeostasis and Muscle Function in Aged C57BL/6J Mice

Obesity exacerbates the age-related decline in muscle function and metabolic health. Despite this, medical treatments for sarcopenic obesity remain limited to lifestyle and behavioral modifications such as calorie restriction and exercise. Here, we hypothesized that sarcopenic obesity can be effectively reversed by reducing mitochondrial bioenergetic efficiency. To achieve this, 80-week-old male C57BL/6J mice with obesity were randomized to 10 weeks of either vehicle (V) or BAM15 (B; 0.1% w/w in diet) treatment, an orally available mitochondrial uncoupler. Body weight and food intake were measured weekly. Body composition, muscle function, energy expenditure, locomotor activity, and glucose tolerance were determined after treatment. Blood and organ weights were obtained at necropsy. BAM15 reduced body weight (B:42.3±1.3 vs. V:54.0±1.3 g, P=0.002) by increasing energy expenditure (B:0.012±0.0 vs. V:0.010±0.0 kcal/g/hr, P=0.003) without altering food intake (B:0.67±0.07 vs. V:0.63±0.02 kcal/hr, P=0.58). BAM15 reduced body fat (B:21.3±1.6 vs. V:31.4±0.5%, P=0.0001) while preserving lean mass (B:59.4±1.0 vs. V:52.7±0.4%, P=0.034). BAM15 increased both muscle strength (B:128.0±2.5 vs. V:92.0±2.0 g, P<0.0001) and locomotor activity (B:432.7±32.0vs. V:347.0±14.4 m/hr, P=0.035). Additionally, BAM15 reduced circulating glucose (B:202.7±9.3 vs. V:245.0±6.4 mg/dL, P=0.004), C-peptide (B:0.68±0.09 vs. V:1.60±0.22 mg/dL, P=0.004), and insulin concentrations (B:1.5±0.2 vs. V:2.6±0.3 ng/mL, P=0.02) while modestly improving glucose tolerance (B:236.8±13.1 vs. V:310.0±15.6 2-hr mg/dL, P=0.03). Taken together, these data indicate that reducing mitochondrial coupling efficiency protects against age-related decline in muscle function while improving peripheral metabolic health.

Keratinocyte‐associated Protein 3 , a Novel Adiposity Gene, May Act Through the Ileum

Obesity poses a significant health burden in the United States, and investigating obesity-related genes remains an important area of research. A previous study from our lab used outbred heterogeneous stock (HS) rats to identify Keratinocyte-associated protein 3, (Krtcap3)as a candidate gene within a quantitative trait locus for adiposity. HS rats with the Wistar-Kyoto (WKY) haplotype at this locus had increased liver expression of Krtcap3, which correlated with decreased retroperitoneal fat mass (RetroFat) and decreased serum triglycerides. We developed an in vivo whole-body knock-out rat model of Krtcap3 in the inbred WKY rat using CRISPR/Cas9. Experimental wild-type (WT) and knock-out (KO) rats were placed onto a high-fat (HFD; 60% kcal fat) or low-fat diet (LFD; 10% kcal fat) at six weeks of age and were maintained on the diet for 13 weeks, with weekly measures of body weight and food intake. We hypothesized that KO rats would have increased fat mass compared to WT rats due to altered liver lipid pathways. In an initial pilot study, we found that male and female KO rats have significantly increased body weight at six weeks of age. At the end of the study, KO females—but not males—had a significantly greater fat mass compared to WT. KO females on HFD, but not LFD, had increased food consumption. We analyzed serum and liver lipids between WT and KO rats, and found no significant genotype-driven differences. This indicated that the KO did not alter liver lipid pathways, suggesting that Krtcap3 is affecting fat mass through a tissue other than liver. Previous work has shown that WKY rats exhibit increased liver Krtcap3 relative to the other HS founder strains, but expression in other tissues has not been investigated. We hypothesize that WKY rats will have a higher Krtcap3 expression compared to the other founder strains in the tissue of action of Krtcap3. We have investigated expression patterns in the following tissues in WKY, Fischer 344, and Brown Norway rats: white and brown adipose tissue, liver, heart, kidney, skeletal muscle, ileum, colon, arcuate nucleus, and nucleus accumbens. Krtcap3 expression is significantly higher only in the WKY liver and ileum. Coupled with our food intake phenotype from the pilot study, we propose that Krtcap3 may be participating in the gut-brain axis to alter food intake and influence adiposity. Future studies will evaluate expression patterns in additional inbred strains and investigate how gut hormones are altered in the KO.

Crtc1 Deficiency Causes Obesity Potentially via Regulating PPARγ Pathway in White Adipose.

Obesity is characterized by excessive fat accumulation and associated with glucose and lipid metabolism disorders. Crtc1, a transcription cofactor regulating CREB activity, has been involved in the pathogenesis of metabolic syndrome; however, the underlying mechanism remains under debate. Here we generated a Crtc1–/– mouse line using the CRISPR/Cas9 system. Under normal feeding conditions, Crtc1–/– mice exhibited an obese phenotype resultant from the abnormal expansion of the white adipocytes. The development of obesity in Crtc1–/– mice is independent of alterations in food intake or energy expenditure. Moreover, Crtc1–/– mice were more prone to insulin resistance and dyslipidemia, as evidenced by higher levels of plasma glucose, insulin and FABP4 than wildtype mice. Transcriptome analysis in liver and epididymal white adipose tissue (eWAT) showed that the fat accumulation caused by Crtc1 deletion was mainly related to lipid metabolism in adipose tissue, but not in liver. GSEA and KEGG analysis identified PPAR pathway to be of the highest impact on lipid metabolism in eWAT. This regulation was independent of a direct interaction between CRTC1 and PPARγ. Our findings demonstrate a crucial role of Crtc1 in regulating lipid metabolism in adipose during development, and provide novel insights into obesity prevention and therapeutics.

Patterns of Eating Associated with Sleep Characteristics: A Pilot Study among Individuals of Mexican Descent at the US-Mexico Border

ABSTRACT Introduction: Previous studies have linked sleep to risk of diabetes and obesity, at least partially via alterations in food intake. Diabetes and obesity are common among Hispanics/Latinos, and studies are needed to better clarify the role of sleep in health among this group. Utilizing the revised TFEQ-R-18, this study will examine whether eating behaviors such as cognitive restraint, emotional eating and uncontrolled eating are related to self-reported sleep experiences. Specifically, we hypothesized that poor eating habits would be associated with (1) more insomnia symptoms, (2) overall worse sleep quality, (3) increased daytime sleepiness, and (4) shorter sleep duration. Methods: Data were collected from N = 100 adults (age 18–60, 47% female) of Mexican descent in the city of Nogales, AZ (34% not born in the US). Surveys were presented in English or Spanish. Eating Patterns were assessed with the Three-Factor Eating Questionnaire (TFEQ), which resulted in a total score and subscales for “cognitive restraint,” “uncontrolled eating,” and “emotional eating.” Insomnia was assessed with the use of the Insomnia Severity Index (ISI), Sleepiness with the use of the Epworth Sleepiness Scale (ESS), Sleep quality with the use of the Pittsburgh Sleep Quality Index (PSQI), and weekday and weekend sleep duration with the use of the Sleep Timing Questionnaire (STQ). Covariates included age, sex, Body Mass Index (BMI), education and immigrant status. Results: Overall TFEQ score (problematic eating) was positively associated with greater insomnia, poorer sleep quality, more sleepiness, and less weekend (but not weekday) sleep. Mean TFEQ score in the sample was 18.7 (range 0–51). In adjusted analyses, every point on the TFEQ was associated with 0.6 ISI points, 0.8 PSQI points, 0.5 ESS points, and 1.1 minutes of less weekend sleep duration. Regarding subscale scores, relationships were generally seenbetween sleep and emotional eating and unrestricted eating, and not cognitive restraint. Conclusions: Greater insomnia, poorer sleep quality, increased daytime sleepiness and decreased weekend sleep duration were associated with eating patterns at the US-Mexico border, particularly in the area of unrestricted eating and emotional eating. This suggests possible mechanisms linking sleep and obesity in Hispanic/Latinos.

Glutamatergic plasticity within neurocircuits of the dorsal vagal complex and the regulation of gastric functions.

The meticulous regulation of the gastrointestinal (GI) tract is required for the coordination of gastric motility and emptying, intestinal secretion, absorption, and transit as well as for the overarching management of food intake and energy homeostasis. Disruption of GI functions is associated with the development of severe GI disorders and the alteration of food intake and caloric balance. Functional GI disorders as well as the dysregulation of energy balance and food intake are frequently associated with, or result from, alterations in the central regulation of GI control. The faithful and rapid transmission of information from the stomach and upper GI tract to second-order neurons of the nucleus of the tractus solitarius (NTS) relies on the delicate modulation of excitatory glutamatergic transmission, as does the relay of integrated signals from the NTS to parasympathetic efferent neurons of the dorsal motor nucleus of the vagus (DMV). Many studies have focused on understanding the physiological and pathophysiological modulation of these glutamatergic synapses, although their role in the control and regulation of GI functions has lagged behind that of cardiovascular and respiratory functions. The purpose of this review is to examine the current literature exploring the role of glutamatergic transmission in the DVC in the regulation of GI functions.

Mitochondrial uncoupler SHC517 reverses obesity in mice without affecting food intake

AimsMitochondrial uncouplers decrease caloric efficiency and have potential therapeutic benefits for the treatment of obesity and related metabolic disorders. Herein we investigate the metabolic and physiologic effects of a recently identified small molecule mitochondrial uncoupler named SHC517 in a mouse model of diet-induced obesity. MethodsSHC517 was administered as an admixture in food. The effect of SHC517 on in vivo energy expenditure and respiratory quotient was determined by indirect calorimetry. A dose-finding obesity prevention study was performed by starting SHC517 treatment concomitant with high fat diet for a period of 12 days. An obesity reversal study was performed by feeding mice western diet for 4 weeks prior to SHC517 treatment for 7 weeks. Biochemical assays were used to determine changes in glucose, insulin, triglycerides, and cholesterol. SHC517 concentrations were determined by mass spectrometry. ResultsSHC517 increased lipid oxidation without affecting body temperature. SHC517 prevented diet-induced obesity when administered at 0.05% and 0.1% w/w in high fat diet and reversed established obesity when tested at the 0.05% dose. In the obesity reversal model, SHC517 restored adiposity to levels similar to chow-fed control mice without affecting food intake or lean body mass. SHC517 improved glucose tolerance and fasting glucose levels when administered in both the obesity prevention and obesity reversal modes. ConclusionsSHC517 is a mitochondrial uncoupler with potent anti-obesity and insulin sensitizing effects in mice. SHC517 reversed obesity without altering food intake or compromising lean mass, effects that are highly sought-after in anti-obesity therapeutics.

Ginger (Zingiber officinale) Attenuates Obesity and Adipose Tissue Remodeling in High-Fat Diet-Fed C57BL/6 Mice.

Obesity is characterized by excessive fat accumulation in adipose tissue, which is an active endocrine organ regulating energy metabolism. Ginger (Zingiber officinale) is known to have antioxidant, anti-inflammatory, and antiobesity effects, but the role of ginger in modulating adipocyte metabolism is largely unknown. In this study, we hypothesized that ginger supplementation inhibits high-fat (HF)-diet-mediated obesity. C57BL/6 male mice were randomly assigned to three diets for 7 weeks: low fat (LF, 16% kcal from fat), HF (HF, 60% kcal from fat), or HF with 5% ginger powder in diet (HF + G). The HF diet increased body weight (BW) and BW gain, as well as fasting glucose, total cholesterol, and hepatic lipid levels, compared to the LF diet-fed group. Ginger supplementation significantly improved HF-diet-induced BW gain, hyperglycemia, hypercholesterolemia, and hepatic steatosis without altering food intake. Next, we investigated whether ginger modulates adipocyte remodeling. HF-mediated adipocyte hypertrophy with increased lipogenic levels was significantly improved by ginger supplementation. Furthermore, the HF+G group showed high levels of the fatty-acid oxidation gene, carnitine palmitoyltransferase 1 (CPT1), which was accompanied by a reduction in adipocyte inflammatory gene expression. Taken together, our work demonstrated that ginger supplementation attenuated HF-diet-mediated obesity and adipocyte remodeling in C57BL/6 mice.

Gastric motor dysfunction coincides with the onset of obesity in rats fed with high-fat diet.

Exposure to a high-fat diet (HFD) has been reported to impair central autonomic and enteric neurocircuitries, however, the relevant mechanisms and their time course are inadequately clarified. This study aimed to investigate the effects of HFD consumption through the period of adolescence on gastric motor functions in adulthood. Male Sprague-Dawley rats consumed a regular diet or HFD (60% kcal by fat) from 4 to 12 weeks of age. Body weight and food intake were monitored weekly. In adult rats, gastric emptying (GE) was measured. Additionally, using in-vitro organ bath, contractile and relaxant responses of antral and fundic strips were assessed with bethanechol and sodium nitroprusside (SNP), respectively. The expressions of choline acetyltransferase (ChAT), neuronal nitric oxide synthase (nNOS) and vasoactive intestinal polypeptide (VIP) were detected by immunofluorescence, whereas, the number of myenteric neurons were evaluated by staining with cuprolinic blue and enteric neuronal marker PGP 9.5. In adulthood, the HFD did not alter food intake, while significantly increasing the body weight. In HFD-fed adult rats, increased visceral fat mass was accompanied by delayed GE. Moreover, bethanechol- and SNP-induced responses were attenuated in antral and fundic tissues. HFD remarkably decreased the number of myenteric neurons and NOS immunoreactivity both in fundus and antrum. HFD remarkably decreased ChAT expression, while increasing the immunoreactivity for VIP in antrum. In conclusion, consumption of HFD between early adolescence and adulthood results in obesity and impairment of gastric motor functions. Particularly, HFD-induced gastric dysmotility appears to be predominantly dependent on the modifications in the non-adrenergic non-cholinergic inhibitory neurotransmission.

Mid-gestation low-dose LPS administration results in female-specific excessive weight gain upon a western style diet in mouse offspring

Gestational complications, including preeclampsia and gestational diabetes, have long-term adverse consequences for offspring’s metabolic and cardiovascular health. A low-grade systemic inflammatory response is likely mediating this. Here, we examine the consequences of LPS-induced gestational inflammation on offspring’s health in adulthood. LPS was administered to pregnant C57Bl/6J mice on gestational day 10.5. Maternal plasma metabolomics showed oxidative stress, remaining for at least 5 days after LPS administration, likely mediating the consequences for the offspring. From weaning on, all offspring was fed a control diet; from 12 to 24 weeks of age, half of the offspring received a western-style diet (WSD). The combination of LPS-exposure and WSD resulted in hyperphagia and increased body weight and body fat mass in the female offspring. This was accompanied by changes in glucose tolerance, leptin and insulin levels and gene expression in liver and adipose tissue. In the hypothalamus, expression of genes involved in food intake regulation was slightly changed. We speculate that altered food intake behaviour is a result of dysregulation of hypothalamic signalling. Our results add to understanding of how maternal inflammation can mediate long-term health consequences for the offspring. This is relevant to many gestational complications with a pro-inflammatory reaction in place.

Leptin alters energy intake and fat mass but not energy expenditure in lean subjects

Based on studies in mice, leptin was expected to decrease body weight in obese individuals. However, the majority of the obese are hyperleptinemic and do not respond to leptin treatment, suggesting the presence of leptin tolerance and questioning the role of leptin as regulator of energy balance in humans. We thus performed detailed novel measurements and analyses of samples and data from our clinical trials biobank to investigate leptin effects on mechanisms of weight regulation in lean normo- and mildly hypo-leptinemic individuals without genetic disorders. We demonstrate that short-term leptin administration alters food intake during refeeding after fasting, whereas long-term leptin treatment reduces fat mass and body weight, and transiently alters circulating free fatty acids in lean mildly hypoleptinemic individuals. Leptin levels before treatment initiation and leptin dose do not predict the observed weight loss in lean individuals suggesting a saturable effect of leptin. In contrast to data from animal studies, leptin treatment does not affect energy expenditure, lipid utilization, SNS activity, heart rate, blood pressure or lean body mass.

Cardiac Autophagy Deficiency Attenuates ANP Production and Disrupts Myocardial-Adipose Cross Talk, Leading to Increased Fat Accumulation and Metabolic Dysfunction.

Increased myocardial autophagy has been established as an important stress-induced cardioprotective response. Three weeks after generating cardiomyocyte-specific autophagy deficiency, via inducible deletion of autophagy-related protein 7 (Atg7), we found that these mice (AKO) had increased body weight and fat mass without altered food intake. Glucose and insulin tolerance tests indicated reduced insulin sensitivity in AKO mice. Metabolic cage analysis showed reduced ambulatory activity and oxygen consumption with a trend of elevated respiratory exchange ratio in AKO mice. Direct analysis of metabolism in subcutaneous and visceral adipocytes showed increased glucose oxidation and reduced ATGL expression and HSL phosphorylation with no change in lipid synthesis or fatty acid oxidation. Importantly, we found AKO mice had reduced myocardial and circulating levels of atrial natriuretic peptide (ANP), an established mediator of myocardial-adipose cross talk. When normal ANP levels were restored to AKO mice with use of osmotic pump, the metabolic dysfunction evident in AKO mice was corrected. We conclude that cardiac autophagy deficiency alters myocardial-adipose cross talk via decreased ANP levels with adverse metabolic consequences.

Highly Conserved Molecular Features in IgLONs Contrast Their Distinct Structural and Biological Outcomes

Neuronal growth regulator 1 (NEGR1) and neurotrimin (NTM) are abundant cell-surface proteins found in the brain and form part of the IgLON (Immunoglobulin LSAMP, OBCAM, Neurotrimin) family. In humans, NEGR1 is implicated in obesity and mental disorders, while NTM is linked to intelligence and cognitive function. IgLONs dimerize homophilically and heterophilically, and they are thought to shape synaptic connections and neural circuits by acting in trans (spanning cellular junctions) and/or in cis (at the same side of a junction). Here, we reveal homodimeric structures of NEGR1 and NTM. They assemble into V-shaped complexes via their Ig1 domains, and disruption of the Ig1–Ig1 interface abolishes dimerization in solution. A hydrophobic ridge from one Ig1 domain inserts into a hydrophobic pocket from the opposing Ig1 domain producing an interaction interface that is highly conserved among IgLONs but remarkably plastic structurally. Given the high degree of sequence conservation at the interaction interface, we tested whether different IgLONs could elicit the same biological effect in vivo. In a small-scale study administering different soluble IgLONs directly into the brain and monitoring feeding, only NEGR1 altered food intake significantly. Taking NEGR1 as a prototype, our studies thus indicate that while IgLONs share a conserved mode of interaction and are able to bind each other as homomers and heteromers, they are structurally plastic and can exert unique biological action.

Butein inhibits lipogenesis in Caenorhabditis elegans.

Butein, a flavonoid found in annatto seeds and lacquer trees, may be used for many health benefits, including the prevention of obesity. However, its anti-obesity effects are not completely understood; in particular, the effects of butein on the regulation of lipid metabolism have not been explained. Thus, the goal of the current study was to determine the effects of butein on lipid metabolism in Caenorhabditis elegans, which is a multi-organ nematode used as an animal model in obesity research. Butein at 70 μM reduced triglyceride content by 27% compared to the control without altering food intake and energy expenditure. The reduced triglyceride content by butein was associated with the downregulation of sbp-1, fasn-1, and fat-7, the lipogenesis-related homologs of sterol regulatory element-binding proteins, fatty acid synthase and stearoyl-CoA desaturase, respectively. Furthermore, fat-7 and skn-1, a homolog of nuclear respiratory factors, were identified as genetic requirements for butein's effects on triglyceride content in C. elegans. The effects of butein on sbp-1 and fasn-1 were dependent on skn-1, but the downregulation of fat-7 was independent of skn-1. These results suggest that the inhibitory effects of butein on lipogenesis are via SKN-1- and FAT-7-dependent pathways in C. elegans.

Interaction of FTO rs9939609 and the native American-origin ABCA1 p.Arg230Cys with circulating leptin levels in Mexican adolescents diagnosed with eating disorders: Preliminary results

Eating disorders (ED) are characterized by disruption of eating behaviour and alteration of food intake. Leptin, is one of the main hormones that modulate food intake and are altered in individuals diagnosed with ED. Genetic risk variants for obesity, like those reported inFTO and ABCA1, have also been associated to ED disorders. The present study aimed to analysed leptin circulating levels and the interaction between obesity-risk variants in FTO and ABCA1, in adolescents diagnosed with ED. A total of 99 individuals diagnosed with ED were genotype using Taqman probes for FTO (rs9939609) and ABCA1 (p.Arg230Cys, rs9282541). Commercial enzyme-linked immunosorbent assays were utilized to determined circulating leptin. Differences in leptin concentration were analysed by t-Student or ANOVA test. Gene-gene interaction were analysed using general estimation equations. Circulating leptin levels differed between the three diagnostic groups, lead by individuals diagnosed with binge eating-disorder. In individuals with more than 3 of episodes of binge-eating per week having the highest leptin levels. Also, we found that carriers of both risk alleles had the highest leptin levels. Our observations found an interaction between FTO rs9969609 and the native American-origin ABCA1 p.Arg230Cys to modulate circulating leptin levels in Mexican adolescents diagnosed with eating-disorders.

1930-P: The Knockdown of TRIL in POMC-Expressing Neurons Reduces Body Adiposity and Ameliorates Leptin Responsiveness

Chronic HFD intake induces low-grade inflammation in the hypothalamus, which is characterized by proinflammatory cytokine overproduction. This process occurs, at least in part, via TLR4 activation by circulant saturated fatty acids. The TLR4 signaling in the brain is mediated by the TLR4 interactor with leucine-rich repeats (TRIL), an upstream accessory protein. Our main purpose was to determine the putative involvement of TRIL in diet-induced hypothalamic inflammation. We employed male, 8-12 week-old C57BL and POMCCRE mice fed on chow or HFD. The mRNA levels in the hypothalamus were measured by RT-PCR, western blot analyses was used for determination of protein content and immunofluorescence microscopy for evaluate the distribution of TRIL in arcuate nucleus (ARC) of hypothalamus and leptin signaling in POMC-expressing neurons. Finally, bilateral viral injections were performed to non-specific knockdown of TRIL in ARC using shRNA-based lentivirus and Cre-dependent AAV to knockdown TRIL specifically in POMC neurons. The non-specific knockdown of TRIL in the ARC of mice fed on HFD attenuated body weight gain despite no alterations in food intake, improved glucose homeostasis and increased energy expenditure. Moreover, nlrp3, il1β and hspa5 mRNA expression were decreased upon knockdown of TRIL. POMC-specific knockdown of TRIL reduced body fat with no changes in food intake or body weight gain in mice fed on HFD for 8 weeks. The inhibition of TRIL in POMC neurons resulted in a trend of improved leptin signaling in those neurons. We also found increased expression of thermogenic genes in the iBAT and improved fasting glucose. POMC-specific inhibition of TRIL in mice fed on HFD for long-term (24 weeks) did not prevent POMC loss or diminishes cleaved-caspase 3 in ARC. Collectively our results suggest that TRIL is involved in the TLR4-mediated early response to dietary fats and TRIL targeting counteract hypothalamic inflammation and partially restores metabolic homeostasis. Disclosure A. Moura-Assis: None. J. Junior: None. J.M. Gaspar: None. L. Velloso: None. P.S. Nogueira: None. Funding São Paulo Research Foundation (2016/01245-5)

Role of Gut Microbiota in Anti-Colitic Effects of Color-Fleshed Potatoes

Abstract Objectives The prevalence of ulcerative colitis (UC), a chronic inflammatory bowel disease, is on the rise with ∼700,000 patients in the US alone in 2018. Gut bacterial dysbiosis plays an important role in ulcerative colitis. We have recently shown that anthocyanin-containing potatoes exert anti-inflammatory activity in colitic mice. However, no information is available on whether gut bacteria play a role in the anti-colitic activity of color-fleshed potatoes. This study examined the anti-colitic activity of red/purple-fleshed potatoes in mice with intact and antibiotic-ablated microbiome. Methods We used DSS-induced murine (C57BL6) colitis model with and without the administration of antibiotics in drinking water for nine weeks. Mice were randomly assigned to the control (AIN-93G diet), DSS (AIN-93G diet), DSS + PP (20% purple potato) and DSS + RP (20% red potato) groups. After eight weeks, mice were treated with 2% DSS in their drinking water for five days to induce colitis. Intestinal permeability was measured using FITC-dextran. Serum myeloperoxidase (MPO) levels were measured using ELISA. RT-PCR was used to analyze the relative gene expression levels of cytokines and bacterial abundance. Results Administration of antibiotics resulted in a 95% reduction in gut bacterial load. Antibiotics administration did not alter food intake, water intake, and weight gain. Antibiotic-treated mice had five times greater cecum weight, a hallmark of germ-free mice, compared to no-antibiotic mice. In antibiotic mice, DSS-induced splenomegaly, elevated gut permeability (serum levels of FITC-dextran), and reduced colon length and weight were more pronounced compared to no antibiotic mice. Purple- or red-fleshed potato supplementation (20% w/w) ameliorated (P ≤ 0.05) DSS-induced reduction in colon length, elevation in permeability, spleen weight and MPO levels in no antibiotic mice only. Moreover, purple-fleshed potato supplementation alone improved the ZO-1 and MUC-2 gene expression levels in no-antibiotic mice, but not in microbiota-ablated mice. Conclusions In summary, these results suggest that purple-fleshed potatoes are more potent compared to red-fleshed potatoes and the gut microbiome is critical for the anti-colitic activity of anthocyanin-containing potatoes. Funding Sources USDA-NIFA awards 2016-67,017-29,285 and 2019-67,017-29,258.

BDNF Val66Met polymorphism alters food intake and hypothalamic BDNF expression in mice.

Obesity, a rising public health burden, is a multifactorial disease with an increased risk for patients to develop several pathological conditions including type 2 diabetes mellitus, hypertension, and cardiovascular disease. Increasing evidence suggests a relationship between the human brain-derived neurotrophic factor (BDNF) Val66Met single-nucleotide polymorphism (SNP) and obesity, although the underlying mechanisms of this connection are still not completely understood. In the present study, we found that homozygous knock-in BDNFMet/Met mice were overweight and hyperphagic compared to wildtype BDNFVal/Val mice. Increased food intake was associated withreduction of total BDNF and BDNF1, BDNF4 and BDNF6 transcripts in the hypothalamus of BDNFMet/Met mice. Incontrast, in the white adipose tissue total BDNF and Glut4 expression levels were augmented, while sirtuin 1 and leptin receptor (Ob-R) expression levels were reduced in BDNFMet/Met mice. Moreover, plasmatic leptin levels were decreased in BDNFMet/Met mice. However, BDNFVal/Val and BDNFMet/Met mice showed a similar response to the insulin tolerance test and glucose tolerance test. Altogether, these results suggest that BDNF Val66Met SNP strongly contributes to adipose tissue pathophysiology, resulting in reduced circulating leptin levels and hypothalamic expression of BDNF, which, in turn, promote increased food intake and overweight in BDNFMet/Met mice.