Fast Food Diet Research Articles

Thermoneutrality Inhibits Thermogenic Markers and Exacerbates Nonalcoholic Fatty Liver in Mice (P21-068-19)

ObjectivesNonalcoholic fatty liver disease (NAFLD) is the most prevalent chronic liver disease, affecting more than a third of the US population and 25% of the global population. In this study, we investigated whether manipulation of brown adipose tissue (BAT) activity via housing C57BL/6 J mice under two different housing temperatures (22°C vs 27°C) influences the development of NAFLD. MethodsMale C57BL/6 J mice were randomly allocated to 4 groups (8 mice/group). Briefly, mice housed at the standard temperature (ST) (22°C) or thermoneutral temperature (TT) (27°C) were fed either a control chow diet (CHD) (Picolab rodent diet 20, LabDiet) with a regular drinking water or an “fast food” diet (FFD) (Research Diets, D12079B) plus fructose 23.1 g/L and glucose 18.9 g/L added to the drinking water for 10 weeks. ResultsMice under TT had significant less food intake compared to their counter partners under ST, regardless of diets. Mice exposed to TT and fed FFD had the highest body weight and plasma leptin level among all groups. For mice fed CHD, mice at TT did not present hepatic steatosis although they slightly gained more body weight compared to mice at ST. For mice fed FFD, thermoneutral housing mice had greater liver weight, liver triglyceride, and exacerbated hepatic steatosis compared to the standard housing mice. Moreover, for mice fed FFD, TT compared to ST had significantly elevated expression of fatty acid synthase, sterol regulatory element-binding transcription factor 1, fatty acid translocase, and monocyte chemoattractant protein-1 in liver. By contrast, TT did not change expression of above genes in mice fed CHD. Furthermore, thermoneutrally housed mice displayed a decrease in gene expression of thermogenic markers, such as uncoupling protein 1, elongation of very long chain fatty acids 3, and cell death-inducing DNA fragmentation factor alpha-like effector A in BAT and inguinal white adipose tissue (WAT). ConclusionsThermoneutrality inhibits expression of thermogenic markers in both BAT and WAT, which correlated to exacerbated NAFLD in mice. Our data indicate that activating BAT and/or promoting WAT browning may represent a potential strategy for the management of NAFLD. Funding SourcesThe College of Human Sciences of Texas Tech University; the National Center for Complementary & Integrative Health.

1133-P: Four Days of Hyperoxia Exposure Confers Sustained Improvement in Glucose Homeostasis

Obesity and type 2 diabetes are co-morbidities, and their prevalence is continually growing. In obesity, in the face of lipid accumulation, adipose tissue become hypoxic with reduced blood flow, accompanied by low grade inflammation and insulin resistance. We therefore hypothesized that exposure to high oxygen (hyperoxia) could improve glucose homeostasis in obesity. We treated normal and fast-food diet (40% fat)-induced obese mice (10 months old) with hyperoxia (95%) for 4 days and quantified the effects on glucose homeostasis and insulin sensitivity. We found that immediately after the hyperoxic treatment, glycemia was modestly higher compared to pretreatment levels. However, both fasting and post-challenge blood glucose levels were significantly decreased after 6 days of hyperoxia treatment in both normal (28±8%; p=0.01) and obese mice (36±4%; p=0.007)). Plasma insulin levels decreased by 41% in both normal (p=0.0026) and obese (p=0.0033) mice and Homeostasis Model Assessment of Insulin Resistance (HOMA-IR) values were also significantly decreased (63±4%; p=0.0009 in normal and 55±13%; p=0.0049 in obese mice)after hyperoxia treatment, indicating that the improvements in glucose homeostasis were associated with improved insulin sensitivity. Surprisingly, these metabolic improvements were maintained more than one month after hyperoxia treatment, and occurred without significant effects on weight. These results demonstrate that exposure to hyperoxia improves fasting blood glucose levels, glucose tolerance and insulin sensitivity, suggesting that the hyperoxia treatment may provide a new approach for supportive treatment of diabetes and obesity. Disclosure S. Jain: None. A.V. Harrison: None. F. Lorenzo: None. D. McClain: None. Funding U.S. Department of Veterans Affairs; National Institute of Diabetes and Digestive and Kidney Diseases

ROLE OF BODY WEIGHT AND BODY FIGURE IN THE DETERMINATION OF ADOLESCENT FOOD HABITS

Body weight showed non-significant difference for fast food and nutritional diet for 13-15 years old boys and girls adolescents. Body weight of boy’s adolescent increased over normal body weight of 13-15 years. Whereas, lower body weight was observed in 16-18 boys adolescent and 13-15 years and 16-18 years girls where body weight was lower than normal body weight. The general body figure of boys and girls adolescent have nutritional diet except fast food and nutritional food habits in girls of 13-15 yrs and 16-18 yrs girls of nanus body figure respectively. Interestingly, girls of 13-15 yrs, had fast food in diet whose body figure was midget. The t-test indicated that boys and girls body figure had highly significant differences in their food habits in both age groups 13-15 yrs and 16-18 yrs.

OR22-6 Reversal Of Diet Induced Metabolic Syndrome In Mice With An Orally Active Small Molecule Inhibitor Of PAI-1

Plasminogen activator inhibitor 1 (PAI-1) is a member of the serpin family of proteases that inhibits tissue-type plasminogen activator and urokinase. Elevated levels of plasma PAI-1 are one of the hallmarks of obesity and are predictably elevated in patients with the metabolic syndrome and type 2 diabetes mellitus. Beyond these correlative relationships, increased PAI-1 levels are a predictor of, and a potential contributor to the development of obesity and diabetes. In experimental studies, mice lacking PAI-1 are resistant to weight gain and glucose intolerance when fed a high fat diet. We recently demonstrated that humans that carry a heterozygous loss-of-function mutation in the gene that codes for PAI-1 (SERPINE1) have lower fasting insulin levels and are protected from the development of diabetes compared with unaffected individuals in the same kindred. In this study, we tested the effects of the novel PAI-1 inhibitor TM5614 in reversing diet-induced obesity and glucose intolerance. Mice were fed a fast food diet for 4 months and treated with TM5614 for 10 days at the conclusion of the study period. After 1 week of treatment, the mice exhibited improvement in several metabolic measurements, including fasting glucose (from 131.2 to 92.7 mg/dL, p<0.0001), fasting plasma insulin (from 0.507 to 0.3722 ng/mL, p=0.0278) and fasting plasma LDL (from 165.1 to 118 mg/dL, p<0.0296). Mice treated with TM5614also exhibited a remarkable reduction in liver steatosis. In order the potentially identify mechanisms to explain these finding, RNA-seq of liver tissue was performed on hepatic tissue from mice treated with TM5614. The two most statistically significant changes were seen in PCSK9 (log2 fold change = -2.8 p = 3.97X10-36) and FGF21 (log2 fold change = 2.5, p = 1.03X10-31) These data suggest that suggest that PAI-1 is directly involved in the pathogenesis of not only T2DM, but also in nonalcoholic fatty liver disease and that selective inhibitors may be of benefit in the prevention and treatment of obesity and obesity-related disorders, including NASH.

Human amnion epithelial cells and their soluble factors reduce liver fibrosis in murine non-alcoholic steatohepatitis.

Non-alcoholic steatohepatitis (NASH) can lead to cirrhosis and hepatocellular carcinoma. Currently, lifestyle modification is the only effective treatment. We have shown that human amnion epithelial cells (hAECs) reduce inflammation and fibrosis in toxin-induced liver injury models. We examined the effect of these cells and the soluble factors released by the cells into culture medium (hAEC conditioned medium [hAEC-CM]) in a diet-induced murine NASH model. C57BL/6J male mice received a Western "fast food diet" for 42weeks. Group 1 received an intraperitoneal injection of 2×106 hAECs at week 34, group 2 received an additional hAEC dose at week 38, and group 3 received thrice weekly hAEC-CM injections intraperitoneal for 8weeks from week 34. Liver fibrosis area, inflammation, and fibrosis regulators were measured by immunohistochemistry, qPCR, and gelatin zymography. Metabolic parameters were also assessed. Fast food diet-fed mice demonstrated peri-cellular hepatic fibrosis, inflammation, and steatosis typical of NASH. Liver fibrosis area was reduced by 40% in hAEC-treated and hAEC-CM-treated mice. hAEC treatment significantly reduced pSMAD 2/3 signaling and the number of activated hepatic stellate cells and liver macrophages. Matrix metalloproteinase 2 and 9 gene and protein expression were variably affected. hAEC treatment did not alter the NASH activity score or metabolic parameters such as bodyweight, total cholesterol, or glucose tolerance. Human amnion epithelial cell and hAEC-CM significantly reduced hepatic inflammation and fibrosis in a diet-induced non-alcoholic fatty liver disease model. Although hAEC and hAEC-CM did not affect the metabolic components of NASH, their therapeutic potential is promising and warrants further investigation.

Thioacetamide potentiates high cholesterol and high fat diet induced steato-hepatitic changes in livers of C57BL/6J mice: A novel eight weeks model of fibrosing NASH

There is an inadequacy of relevant animal models to study non-alcoholic steatohepatitis (NASH) and fibrosis. Here, we co-administered thioacetamide (TH) along with fast food diet (FFD) to C57BL/6 J mice for eight weeks. The treatments were: a) standard chow, SC b) FFD c) FFD + TH [75 mg/kg], FTH d) SC + TH [150 mg/kg], STH for 8 weeks. In in-vitro model, Hep3B cells were exposed to palmitic acid (PA) and TH viz. PA (0.25 mM) + TH (25 mM), PA (0.5 mM) alone and TH (50 mM) alone for 12 h, later supernatant media was transferred to LX-2 cells, for another 12 h. Molecular and cellular events related to inflammation, fibrosis, collagen deposition were studied. The FTH mice featured hepatic inflammation, severe diffuse fibrosis, and collagen deposition, which were less severe in FF & STH groups. In FTH group the protein expressions of α-SMA, TGF-ß, Col1 A1, CYP2E1, were up-regulated as compared to the FF group. The in-vivo findings were complemented in the LX-2 and Hep3B cells. The protein expressions of inflammatory and cellular injury markers were significantly higher in PA + TH exposed LX-2 cells. This novel model manifested hepatic inflammation and fibrosis in just eight weeks, which may be exploited for rapid screening of novel anti-NAFLD and liver anti-fibrotic agents.

Longitudinal characterization of diet-induced genetic murine models of non-alcoholic steatohepatitis with metabolic, histological, and transcriptomic hallmarks of human patients.

ABSTRACTNon-alcoholic steatohepatitis (NASH) is a fast-growing liver disease in the Western world. Currently, only a few animal models show both the metabolic and histological features of human NASH. We aimed to explore murine NASH models in a time dependent manner that exhibit metabolic, histological and transcriptomic hallmarks of human NASH. For this, the murine strains C57BL/6J, ob/ob, and KK-Ay were used and three types of nutritional regimes were administered: normal chow diet (NCD); high-fat, high-fructose, and high-cholesterol diet (fast food diet; FFD); or choline-deficient, L-amino acid-defined, high-fat diet (CDAHFD), for 2, 4, 8, 12, 18, 24, and 30 weeks. All strains under the FFD and CDAHFD regimes developed steatohepatitis. Among the strains treated with FFD, the non-alcoholic fatty liver disease (NAFLD) activity score, fibrosis progression and metabolic abnormalities such as hyperinsulinemia and obesity were more pronounced in ob/ob mice than in C57BL/6J and KK-Ay mice. In ob/ob mice fed FFD, the development of hepatic crown-like structures was confirmed. Furthermore, molecular pathways involved in steatohepatitis and fibrosis showed significant changes from as early as 2 weeks of starting the FFD regime. Ob/ob mice fed FFD showed metabolic, histological, and transcriptomic dysfunctions similar to human NASH, suggesting their potential as an experimental model to discover novel drugs for NASH.

DETERMINATION OF ADOLESCENT FOOD HABITS ON THE BASIS OF AGE, HEIGHT AND WEIGHT

Significant differences between nutritional and fast food habits were observed at the 13-15 years and 16-18 years age groups of boys and girls respectively. The girls in the age group and boys had non-significant difference. Body weight showed non-significant difference for fast food and nutritional diet for boys and girls. Body weight of boys increased by 8.90% over normal body weight of 13-15 years. Lower body weight was observed in 16-18 boys adolescent and 13-15 years and 16-18 years girls where body weight was lower than normal body weight. Fast food did not affect in the enhancement of body weight. Nutritional food had significant role in boy weight increase in both age groups.

Local fast-food environment, diet and blood pressure: the moderating role of mastery

PurposeTo examine the moderating role of mastery in the association of local fast-food restaurants (FFR) with diet quality and systolic blood pressure (SBP).MethodsWe used cross-sectional data from 1543 adults participating in wave six of the Netherlands Study of Depression and Anxiety (NESDA). Data were collected between 2013 and 2016. Diet quality was defined by adherence with the dietary approaches to stop hypertension (DASH) diet. Individuals reported on their food consumption through a food frequency questionnaire and SBP was measured. Density of FFR in 1600 m, 800 m and 400 m circular buffers around the home postal code was calculated using Geographic Information Systems. We assessed the association between density of FFR, diet and SBP using linear regression analyses, testing for moderation by mastery.ResultsMean age was 52 years and 32.2% of the sample were men. Exposure to FFR ranged from 0 to 35 FFR per km2. Density of FFR was not significantly associated with DASH adherence or SBP. Only one out of the six interaction terms was significant, suggesting that for individuals with lower levels of mastery, higher density of FFR in an 800-m buffer was negatively associated with DASH adherence, while for individuals with higher levels of mastery, this association was positive.ConclusionsExposure to FFR was not associated with diet quality and SBP, and we observed little evidence for moderation by level of mastery. This research question should be further explored in a large sample of healthy adults.

Geographical Distribution of Fast Food and CRC Incidence in Young Adults

Background: There is an increasing trend of colorectal cancer (CRC) in young adults who also used to take fast food. However, little is known about the association between fast food and the occurrence of CRC in young adults. Aim: To investigate the impact of fast food diet on the incidence rate of CRC in terms of geographic distribution. Methods: Geographic distribution of fast food diet was measured by the density of fast food stores (numbers of fast food stores/population per 105) according to each county. We summed up total numbers of the fast food stores ranking the top 3 in Taiwan from the corresponding official Web sites. The incidence of CRC from 2004 to 2013 for young adults aged 15 to 49 years old was computed from Interactive Information System of the National Cancer Registry, provided by Health Promotion Administration, Ministry of Health and Welfare. Multilevel random-effect regression model was applied to estimating the effects of geographical location of fast food stores at city/county level on incidence of CRC with adjustment for age and gender at individual level. Results: The incidence rate of CRC in young adults aged 50 years or below was in parallel with the distribution of fast food stores in geographical pattern. After adjusting for age and sex, the results show that an increase in the growth of 1 unit of density in fast food store led to an increase in incidence of CRC by 2%. Approximately 87% accuracy was noted in predicting the highest one-third percentage of incident CRC cases in young adults. Conclusion: The density of fast food stores was highly associated with the increased risk of colorectal cancer incidence in young adults. This may account for the increasing trend of CRC in young adults in Taiwan.

Supervised learning reveals circulating biomarker levels diagnostic of hepatocellular carcinoma in a clinically relevant model of non-alcoholic steatohepatitis; An OAD to NASH.

Although cirrhosis is a key risk factor for the development of hepatocellular carcinoma (HCC), mounting evidence indicates that in a subset of patients presenting with non-alcoholic steatohepatitis (NASH) HCC manifests in the absence of cirrhosis. Given the sheer size of the ongoing non-alcoholic fatty liver disease (NAFLD) epidemic and the dismal prognosis associated with late-stage primary liver cancer there is an urgent need for HCC surveillance in the NASH population. Using serum levels of HCC biomarkers as vectors and biopsy-proven HCC or no HCC as outputs / binary classifier, a supervised learning campaign was undertaken to develop a minimally invasive technique for making a diagnosis of HCC in a clinically relevant model of NASH. Adult mice randomized to control diet or a fast food diet (FFD) were followed for up to 14 mo and serum level of a panel of HCC-relevant biomarkers was compared with liver biopsies at 3 and 14 mo. Both NAFLD Activity Score (NAS) and hepatic hydroxyproline content were elevated at 3 and 14 mo on FFD. Picrosirius red staining of liver sections revealed a filigree pattern of fibrillar collagen deposition with no cirrhosis at 14 mo on FFD. Nevertheless, 46% of animals bore one or more tumors on their livers confirmed as HCC in hematoxylin-eosin-stained liver sections. In this training set, receiver operating characteristic (ROC) curves analysis for serum levels of the HCC biomarkers osteopontin (OPN), alpha-fetoprotein (AFP) and Dickkopf-1 (DKK1) returned concordance-statistic/area under ROC curve of ≥ 0.89. Serum levels of OPN (threshold, 218 ng/mL; sensitivity, 82%; specificity, 86%), AFP (136 ng/mL; 91%; 97%) and DKK1 (2.4 ng/mL; 82%; 81%) diagnostic for HCC were confirmed in a test set comprising mice on control diet or FFD and mice subjected to hepatic ischemia-reperfusion injury. These data suggest that levels of circulating OPN, AFP and DKK1 can be used to make a diagnosis of HCC in a clinically relevant model of NASH.

Nordihydroguaiaretic Acid, a Lignan from Larrea tridentata (Creosote Bush), Protects Against American Lifestyle-Induced Obesity Syndrome Diet-Induced Metabolic Dysfunction in Mice.

To determine the effects of nordihydroguaiaretic acid (NDGA) on metabolic and molecular changes in response to feeding a typical American fast food or Western diet, mice were fed an American lifestyle-induced obesity syndrome (ALIOS) diet and subjected to metabolic analysis. Male C57BL/6J mice were randomly assigned to the ALIOS diet, the ALIOS diet supplemented with NDGA (NDGA+ALIOS), or a control diet and were maintained on the specific diet for 8 weeks. Mice fed the ALIOS diet showed increased body, liver, and epididymal fat pad weight as well as increased plasma alanine transaminase (ALT) and aspartate aminotransferase (AST) levels (a measure of liver injury) and liver triglyceride content. Coadministration of NDGA normalized body and epididymal fat pad weight, ALT and AST levels, and liver triglycerides. NDGA treatment also improved insulin sensitivity but not glucose intolerance in mice fed the ALIOS diet. In mice fed the NDGA+ALIOS diet, NDGA supplementation induced peroxisome proliferator-activated receptor α (PPARα; the master regulator of fatty acid oxidation) and mRNA levels of carnitine palmitoyltransferases Cpt1c and Cpt2, key genes involved in fatty acid oxidation, compared with the ALIOS diet. NDGA significantly reduced liver endoplasmic reticulum (ER) stress response C/EBP homologous protein, compared with chow or the ALIOS diet, and also ameliorated ALIOS diet-induced elevation of apoptosis signaling protein, caspase 3. Likewise, NDGA downregulated the ALIOS diet-induced mRNA levels of Pparg, fatty acid synthase Fasn, and diacylglycerol acyltransferase Dgat2 NDGA treatment of ALIOS-fed mice upregulated the hepatic expression of antioxidant enzymes, glutathione peroxidase 4, and peroxiredoxin 3 proteins. In conclusion, we provide evidence that NDGA improves metabolic dysregulation by simultaneously modulating the PPARα transcription factor and key genes involved in fatty acid oxidation, key antioxidant and lipogenic enzymes, and apoptosis and ER stress signaling pathways.

Fasting and Fast Food Diet Play an Opposite Role in Mice Brain Aging.

Fasting may be exploited as a possible strategy for prevention and treatment of several diseases such as diabetes, obesity, and aging. On the other hand, high-fat diet (HFD) represents a risk factor for several diseases and increased mortality. The aim of the present study was to evaluate the impact of fasting on mouse brain aging transcriptome and how HFD regulates such pathways. We used the NCBI Gene Expression Omnibus (GEO) database, in order to identify suitable microarray datasets comparing mouse brain transcriptome under fasting or HFD vs aged mouse brain transcriptome. Three microarray datasets were selected for this study, GSE24504, GSE6285, and GSE8150, and the principal molecular mechanisms involved in this process were evaluated. This analysis showed that, regardless of fasting duration, mouse brain significantly expressed 21 and 30 upregulated and downregulated genes, respectively. The involved biological processes were related to cell cycle arrest, cell death inhibition, and regulation of cellular metabolism. Comparing mouse brain transcriptome under fasting and aged conditions, we found out that the number of genes in common increased with the duration of fasting (222 genes), peaking at 72h. In addition, mouse brain transcriptome under HFD resembles for the 30% the one of the aged mice. Furthermore, several molecular processes were found to be shared between HFD and aging. In conclusion, we suggest that fasting and HFD play an opposite role in brain transcriptome of aged mice. Therefore, an intermittent diet could represent a possible clinical strategy to counteract aging, loss of memory, and neuroinflammation. Furthermore, low-fat diet leads to the inactivation of brain degenerative processes triggered by aging.

Dietary patterns, their covariates, and associations with severity of depressive symptoms among university students in Lebanon: a cross-sectional study.

The study aims to identify dietary patterns in university students in Lebanon, to determine their associations with socio-demographic, lifestyle, and stress factors, and to assess the link between identified dietary patterns and severity of depressive symptoms, controlling for multiple confounders. Four hundred and fifty-seven private university students were surveyed. Information about dietary intake, socio-demographic and lifestyle factors, physical activity, and depressive symptoms were collected by the 73-item food frequency questionnaire, background questionnaire, International Physical Activity Questionnaire-Short Form, and Patient Health Questionnaire (PHQ-9), respectively. Dietary patterns were identified by exploratory factor analysis. Multiple linear regression analyses were conducted to (1) identify covariates that were independently associated with dietary patterns and (2) examine the independent associations between each of the dietary patterns and severity of depressive symptoms. Five dietary patterns were identified: "traditional Lebanese", "Western fast food", "dairy", "Lebanese fast food" and "fruits". Higher scores of traditional Lebanese and fruits patterns were found to be associated with more meals per day, frequent breakfast consumption, and non-smoking. Higher Western fast food diet scores were shown to be associated with male gender, fewer meals per day, less frequent breakfast consumption, more frequent snacking, and alcohol consumption. Higher scores of the dairy pattern were found to be positively associated with hypertension, non-smoking, and frequency of alcohol consumption. Higher Lebanese fast food pattern scores were found to be associated with higher frequency of meal intake while watching TV and alcohol consumption. None of the five dietary patterns showed a significant association with severity of depressive symptoms after controlling for confounders. Severity of depressive symptoms showed no independent association with the identified dietary patterns.

Ferroportin Expression in Adipocytes Does Not Contribute to Iron Homeostasis or Metabolic Responses to a High Calorie Diet.

Background & AimsIron has an increasingly recognized role in the regulation of adipose tissue function, including the expression of adipokines involved in the pathogenesis of nonalcoholic fatty liver disease. The cellular iron exporter, ferroportin, has been proposed as being a key determinant of adipocyte iron homeostasis.MethodsWe studied an adipocyte-specific ferroportin (Fpn1) knockout mouse model, using an Adipoq-Cre recombinase driven Fpn1 deletion and fed mice according to the fast food diet model of nonalcoholic steatohepatitis.ResultsWe showed successful selective deletion of Fpn1 in adipocytes, but found that this did not lead to increased adipocyte iron stores as measured by atomic absorption spectroscopy or histologically quantified iron granules after staining with 3,3’-diaminobenzidine–enhanced Perls’ stain. Mice with adipocyte-specific Fpn1 deletion did not show dysregulation of adiponectin, leptin, resistin, or retinol-binding protein-4 expression. Similarly, adipocyte-specific Fpn1 deletion did not affect insulin sensitivity during hyperinsulinemic–euglycemic clamp studies or lead to histologic evidence of increased liver injury. We have shown, however, that the fast food diet model of nonalcoholic steatohepatitis generates an increase in adipose tissue macrophage infiltration with crown-like structures, as seen in human beings, further validating the utility of this model.ConclusionsFerroportin may not be a key determinant of adipocyte iron homeostasis in this knockout model. Further studies are needed to determine the mechanisms of iron metabolism in adipocytes and adipose tissue.

Fast food diet-induced non-alcoholic fatty liver disease exerts early protective effect against acetaminophen intoxication in mice

BackgroundAcetaminophen (APAP) is a readily available and safe painkiller. However, its overdose is the most common cause of acute liver injury (ALI). Many predisposing factors contribute to susceptibility to APAP-induced ALI. Non-alcoholic fatty liver disease (NAFLD), the major cause of chronic liver disease, is considered an important predictor of APAP-induced ALI, although the exact mechanism controversial. In this study, we aimed to elucidate the effects of NAFLD on APAP-induced ALI.MethodsTwo groups of mice, normal chow (NC) diet-fed and fast food (FF) diet-fed mice for 14 weeks, were further divided into two subgroups: intraperitoneally injected with either saline (NC-S and FF-S groups) or APAP (NC-A and FF-A groups). Biochemical tests, histological analysis, quantitative PCR, and western blotting were conducted.ResultsAlanine aminotransferase (ALT) level (199.0 ± 39.0 vs. 63.8 ± 7.4 IU/L, p < 0.05) and NAFLD activity score (0 vs. 4.5 ± 0.22) were significantly higher in mice in FF-S group than those in NC-S group. ALI features such as ALT level (8447.8 ± 1185.3 vs. 836.6 ± 185.1 IU/L, p < 0.001) and centrizonal necrosis were prominent and mRNA levels of Trib3 (RR, 1.81) was high in mice in the NC-A group. Levels of CYP2E1 and anti-inflammatory molecules such as PPAR-γ, p62, and NRF2 were high in mice in the FF-A group.ConclusionsOur results showed that while the FF diet clearly induced non-alcoholic steatohepatitis and metabolic syndrome, NAFLD also attenuates APAP-induced ALI by inducing anti-inflammatory molecules such as PPAR-γ.

Abstract 64: HDL Glycosylation is Associated with HDL’s Anti-Inflammatory Function and is Responsive to Dietary Intervention

Although HDL cholesterol (HDL-C) levels are protective against cardiovascular disease (CVD) risk, raising HDL-C pharmaceutically has not led to improvements in cardiovascular outcomes. It is becoming clear that HDL composition and function are more important than HDL concentrations in determining risk. This study set out to determine whether glycosylation differences in HDL-associated glycoproteins affect HDL’s anti-inflammatory function. HDL were purified from healthy subjects (n=10), who consumed in randomized order a fast food (FF) diet and a Mediterranean (Med) diet for 4 days, with a 4-day washout between arms. All foods were provided in this isocaloric cross-over intervention trial, which was approved by the UC Davis IRB. Concentrations of HDL-bound serum amyloid A (SAA), apolipoprotein A-I (ApoA-I), apolipoprotein C-III (ApoC-III), α-1-antitrypsin (A1AT), and α-2-HS-glycoprotein (A2HSG); and the site-specific glycosylation of ApoC-III, A1AT, and A2HSG were measured. Secretion of TNF-α in stimulated monocytes was measured to assess HDL anti-inflammatory function. HDL glycosylation was altered by the dietary interventions and correlated with changes in the amount of TNF-α secreted by stimulated monocytes. HDL glycosylation profiles were different in response to the FF diet vs. the Med diet. HDL with a diminished capacity to suppress TNF-α secretion were enriched in ApoC-III and desialylated A2HSG, depleted in A1AT, and had lower levels of sialylation across glycoproteins. Our results demonstrate that HDL glycoprotein composition, including site-specific glycosylation, is responsive to dietary intervention and correlates with HDL’s ability to modulate TNF-α response in stimulated monocytes. These data suggest that the measurement of HDL glycosylation profiles may be useful in stratifying CVD risk and detecting individuals with impaired HDL anti-inflammatory function.

MiR-21 ablation and obeticholic acid ameliorate nonalcoholic steatohepatitis in mice

microRNAs were recently suggested to contribute to the pathogenesis of nonalcoholic fatty liver disease (NAFLD), a disease lacking specific pharmacological treatments. In that regard, nuclear receptors are arising as key molecular targets for the treatment of nonalcoholic steatohepatitis (NASH). Here we show that, in a typical model of NASH-associated liver damage, microRNA-21 (miR-21) ablation results in a progressive decrease in steatosis, inflammation and lipoapoptosis, with impairment of fibrosis. In a complementary fast food (FF) diet NASH model, mimicking features of the metabolic syndrome, miR-21 levels increase in both liver and muscle, concomitantly with decreased expression of peroxisome proliferator-activated receptor α (PPARα), a key miR-21 target. Strikingly, miR-21 knockout mice fed the FF diet supplemented with farnesoid X receptor (FXR) agonist obeticholic acid (OCA) display minimal steatosis, inflammation, oxidative stress and cholesterol accumulation. In addition, lipoprotein metabolism was restored, including decreased fatty acid uptake and polyunsaturation, and liver and muscle insulin sensitivity fully reinstated. Finally, the miR-21/PPARα axis was found amplified in liver and muscle biopsies, and in serum, of NAFLD patients, co-substantiating its role in the development of the metabolic syndrome. By unveiling that miR-21 abrogation, together with FXR activation by OCA, significantly improves whole body metabolic parameters in NASH, our results highlight the therapeutic potential of nuclear receptor multi-targeting therapies for NAFLD.

Short Term Fast Food and Mediterranean Diet are able to Change the HDL lipidome with Different Patterns

HDL (high density lipoprotein) is the key mediator in reverse cholesterol transport. Recent research has also shown that it plays a role in immune modulation. Diet has a significant impact on the HDL lipidome, which is associated with HDL's function. In this present study, we examined the effects of two common diets, the fast food (FF) diet and the Mediterranean (Med) diet, on the HDL lipidome. Five male and five female (n=10) were recruited to participate in a randomized order cross‐over study. Subjects were healthy, non‐smokers, age 18–65, BMI 18.5–25. Each subject was randomized to consume either the Med or FF diet for 4 days in the first treatment period, followed by a 7‐day wash‐out, followed by the other diet for 4 days in the second treatment period. All food was provided to the subjects during the Med diet, and during the FF diet period subjects were given gift cards to purchase specific meals at fast food restaurants. Fasting blood draw was taken before and after each dietary intervention. HDL was isolated using a density‐based sequential ultra centrifugation method, using density 1.63 – 1.21 g/ml. HDL was then extracted using an MTBE‐methanol method and injected into a LC‐QTOF for lipidomic analysis. Feature annotation was done using MS‐DIAL. Quantification was done using lipid class‐specific standard curves. HDL cholesterol efflux capacity was measured using a commercial kit with THP monocytes transformed to macrophages. The Med diet and FF diet differentially altered the HDL lipidome. The FF diet increased lysophospholipids and triglycerides, and fatty acid saturation, while more unsaturated lipids were observed after the Med diet. These change in the HDL lipidome were correlated with cholesterol efflux capacity.

Long-term administration of tacrolimus and everolimus prevents high cholesterol-high fructose-induced steatosis in C57BL/6J mice by inhibiting de-novo lipogenesis

AimTo investigate the effects of tacrolimus (TC) and everolimus (EV) on non-alcoholic steatohepatitis (NASH) induced by high fat, high cholesterol and fructose (fast food) diet in C57BL/6J mice.Materials and MethodsC57BL/6J mice were divided into four groups (n=8). 1) Standard Chow (SC); 2) Fast food (FF) diet; 3) FF + Tacrolimus (TC, 1mg/kg) and; 4) FF + Everolimus (EV, 1mg/kg) and treated for 16 weeks. Serum and tissue samples were analyzed for evidence of inflammation, fibrosis, lipogenesis, and apoptosis.ResultsTC and EV treatments significantly reduced the hepatic lipid accumulation, improved liver-body weight ratio, blood biochemistry, and insulin resistance in mice fed with FF diet. However, inflammation, enlarged portal tracts, and fibrosis were pronounced in EV treated group. The lipogenic parameters, Peroxisome proliferator-activated receptor gamma (PPAR-γ), Sterol regulatory element-binding protein 1(SREBP-1), mammalian target of rapamycin (m-TOR), Stearoyl-CoA desaturase-1 (SCD-1) and fatty acid translocase (CD36) were significantly down-regulated in livers of TC and EV treated groups as compared to FF group. TC improved Bcl2/Bax ratio, decreased apoptosis, CYP2E1 protein expression and liver fibrosis levels, however, EV offered no such protection. Further, in an In-vitro model of lipotoxicity using the mouse hepatocyte (AML-12) cell line, treatment with TC and EV significantly reduced lipid accumulation and lipogenic and apoptotic markers induced with palmitic acid.ConclusionIn FF diet induced model of NASH, both TC and EV inhibited hepatic lipid accumulation and improved metabolic parameters such as insulin resistance and dyslipidemia. However, mice administered with EV exhibited inflammatory and fibrotic responses despite reduced hepatic steatosis.