Fat Feeding Research Articles

Hyperinsulinemic Compensation for Insulin Resistance Occurs Independent of Elevated Glycemia in Male Dogs.

Insulin resistance engenders a compensatory increase in plasma insulin. Inadequate compensation is a primary element in the pathogenesis of type 2 diabetes. The signal that heralds developing insulin resistance and initiates hyperinsulinemic compensation is not known. It has often been assumed to be increased glucose. We tested this assumption by determining whether development of fasting and/or glucose-stimulated hyperinsulinemia with diet-induced insulin resistance occurs because of concomitant elevation of glycemia. Male dogs (n = 58) were fed a hypercaloric, fat-supplemented diet for 6 weeks. Dogs underwent magnetic resonance imaging to quantify total and regional (visceral, subcutaneous) adiposity as well as euglycemic hyperinsulinemic clamps. A subset of animals also underwent an insulin-modified intravenous glucose tolerance test to assess insulin sensitivity, acute insulin response (AIRg), and glucose effectiveness. Fat feeding caused modest weight gain, increased visceral and subcutaneous fat, and insulin resistance at both peripheral and hepatic levels. Hyperinsulinemic compensation was observed in fasting levels as well as increased AIRg. However, we observed absolutely no increase in carefully measured fasting, evening (6 to 8 pm) or nocturnal glycemia (2 to 4 am). Insulin resistance and hyperinsulinemia occurred despite no elevation in 24-hour glucose. Compensatory development of hyperinsulinemia during diet-induced insulin resistance occurs without elevated fasting or 24-hour glycemia. These data refute the idea that glucose itself is a requisite signal for β-cell upregulation. Alternative feedback mechanisms need to be identified.

Cellular and physiological circadian mechanisms drive diurnal cell proliferation and expansion of white adipose tissue

Hyperplastic expansion of white adipose tissue (WAT) relies in part on the proliferation of adipocyte precursor cells residing in the stromal vascular cell fraction (SVF) of WAT. This study reveals a circadian clock- and feeding-induced diurnal pattern of cell proliferation in the SVF of visceral and subcutaneous WAT in vivo, with higher proliferation of visceral adipocyte progenitor cells subsequent to feeding in lean mice. Fasting or loss of rhythmic feeding eliminates this diurnal proliferation, while high fat feeding or genetic disruption of the molecular circadian clock modifies the temporal expression of proliferation genes and impinges on diurnal SVF proliferation in eWAT. Surprisingly, high fat diet reversal, sufficient to reverse elevated SVF proliferation in eWAT, was insufficient in restoring diurnal patterns of SVF proliferation, suggesting that high fat diet induces a sustained disruption of the adipose circadian clock. In conclusion, the circadian clock and feeding simultaneously impart dynamic, regulatory control of adipocyte progenitor proliferation, which may be a critical determinant of adipose tissue expansion and health over time.

202-LB: Reduced Glucose Effectiveness Exacerbates Impaired Glucose Tolerance in Prediabetic Dogs

The prodromal prediabetic state is characterized by glucose intolerance. Both insulin-dependent (sensitivity, secretion, clearance) and -independent factors (“glucose effectiveness”) contribute to glucose homeostasis. The relative importance of glucose effectiveness is not known. Healthy dogs (n=18) were tested at baseline (BL), after 6 wk on a high-fat diet (HFD), and 2 wk, after a sub-diabetogenic dose of streptozotocin (STZ, 18.5 mg/kg); HFD was continued. We performed intravenous glucose tolerance tests (IVGTTs)at 0, 6, and 8 wks. Minimal model analysis quantified insulin sensitivity (SI), acute insulin response to glucose (AIRg), disposition index (DI, = SI x AIRg), and glucose tolerance (KG). Glucose effectiveness (SG) was measured both by minimal model analysis and directly by multi-step hyperglycemic clamps at basal insulin in a subset of the dogs (n=5). Consumption of HFD induced 7.7±1.0% weight gain (P<0.001) and reduced SI (BL: 3.33±0.35; HFD: 2.01±0.38 x 104 min-1 per µU/ml; P=0.02). HFD elicited an increased AIRg (BL: 596±42; HFD: 1865±476 µU/ml, P=0.03) while maintaining KG (BL: 3.33±0.25; HFD: 3.67±0.33 min-1, P=0.69) as well as β-cell function expressed as DI (BL: 1939±236; HFD: 1882±199, P=0.97). SG was not changed by fat feeding per se (P=0.88). Prediabetes (STZ) conserved fasting glucose, but KG deteriorated (-45% from HFD to STZ) due to sustained insulin resistance with compromised β-cell function (DI decreased by 45±8%, P<0.01). Additionally, minimal model-based SG declined 37±8% from HFD to STZ (0.044±0.003 vs. 0.027±0.003 min-1, P<0.01), a result confirmed by glucose clamps (-30%). Impairments in both insulin-dependent and insulin-independent factors contribute to the glucose intolerance of experimental prediabetes. Rather than compensating for insulin resistance and beta-cell dysfunction, glucose effectiveness also declines, demonstrating that it exacerbates and contributes to glucose intolerance in the canine model of prediabetes. Disclosure C. M. Kolka: Research Support; Self; AstraZeneca. J. Porter: None. M. Ader: None. R. N. Bergman: None. Funding National Institutes of Health (DK27619, DK29867)

1139-P: CD36/FABP4/PPAR-d Axis-Induced Inflammation Is Dependent on ACSL1 under the Influence of Acute High-Fat Feeding

Elevated levels of circulating inflammatory macrophages are associated with several metabolic syndrome (MetS) such as obesity and type II diabetes (T2D). The ingestion of high-fat meals was proven by several groups to act as a stimulus for systemic inflammatory responses via activation of peroxisome proliferator-activated receptors (PPARs). However, little is known about the lipid metabolism involved in macrophage-mediated inflammation under the influence of acute high fat feeding (AHFF). ACSL1 is a member of the long chain acyl-CoA synthetase (ACSL) family that plays key roles in fatty acid metabolism in various tissues. Our previous studies showed that attenuation of ACSL1 was sufficient to dampen inflammatory responses in macrophages challenged with TNF-α or LPS. In this study, we investigated the molecular mechanisms underlying AHFF-induced ACSL1 gene transcription in macrophages. In our work, we showed that four hours feeding of palmitic acid induced significant up-regulation of ACSL1 at both gene and protein levels. Both the inhibition and the deficiency of ACSL1, showed a significant impact on downregulating AHFF-induced inflammatory responses. Furthermore, the inhibition ACSL1 reduced mRNA levels of fatty acid uptake-related genes cluster of differentiation 36 (CD36) and fatty acid transport protein 4 (FATP4). However, inhibition of ACSL1 had no impact on lipogenesis related genes; SREBS, FAS and ACACA or fatty acid-oxidation genes; CPT-1a, CPT-1b and CPT2. Our data also showed that ACSL1 inhibition reduced PPARδ gene expression, while PPARα and PPARγ were not affected. Of note, macrophage intracellular lipid accumulation was attenuated during ACSL inhibition. Our findings suggest that targeting ACSL1 has a therapeutic potential to prevent AHFF-induced macrophage inflammation. Disclosure F. Alrashed: None. A. Al madhoun: None. S. T. Sindhu: None. A. Al-sayyar: None. F. Almulla: None. R. Ahmad: None. Funding Kuwait Foundation for the Advancement of Sciences (CB-2020-004)

1206-P: Effects of Short-Term Administration of Delta-9-tetrahydrocannabinol on Metabolic Parameters of C57bl/6 Mice

Cancer and HIV-related cachexias are among the qualifying conditions for medical cannabis use. Although effective to alleviate nausea and vomiting, clinical trials demonstrated little to no impact of cannabinoids on weight gain in patients. Consistent with this, cannabis users have a lower prevalence of obesity in epidemiological studies. Since cannabinoid-1 receptors (CB1R) regulate food intake and adipose tissue thermogenesis, we studied the metabolic effects of a low dose of delta-9-tetrahydrocannabinol (THC), a CB1R agonist and the main bioactive chemical in cannabis, in 8-week old male mice maintained on a 10%-fat-calorie diet. Mice received daily intraperitoneal injections of THC (3 mg/Kg, n = 15) or vehicle (n = 12) just before the dark cycle for 7 days. THC-treated mice gained an average of 1.6 grams compared to 2.6 grams gained by the vehicle-treated mice (p = 0.03). Despite the attenuated weight gain, automated home cage phenotyping revealed an increase in food intake in the 90 minutes after injections in the THC-treated mice (mean = 0.78 vs. 0.53 grams; p = 0.001). No differences, however, were detected in overall light (p = 0.39) or dark-cycle food intake (p = 0.07). THC-treated mice also showed reduced locomotor activity in the dark cycle compared to vehicle-treated mice (p <0.001). There were no robust differences in indirect calorimetry parameters between groups in the dark cycle, whereas the respiratory exchange rates were lower among THC-treated mice during the light cycle (p = 0.01). Gene expression studies revealed reduced CB1R mRNA levels in brown adipose (mean = 0.61; p = 0.02) and liver (mean = 0.30; p = 0.04) of THC-treated mice. In summary, administration of a low dose of THC was associated with attenuated weight gain in male C57BL/6 mice, possibly through downregulation of CB1R in metabolically active tissues. Future studies will elucidate the effects of chronic THC administration on CB1R expression and thermogenesis under high fat feeding and obesity. Disclosure E. Kurjan: n/a. L. K. Olson: None. O. Alshaarawy: None. Funding National Institutes of Health (R00AT009156)

284-OR: Hepatocyte-Specific CCN2 Gene Deletion Prevents NASH Fibrosis in a Mouse Model of High-Fat Feeding and Diabetes

Introduction: CCN2, formerly known as CTGF, is a pro-fibrotic protein that is implicated in several diabetes-related complications. The role of CCN2 in mediating NASH fibrosis in diabetes has not been fully elucidated. We hypothesize that targeting liver CCN2 can prevent NASH fibrosis in diabetes. Aim: To examine whether hepatocyte specific homozygous gene deletion of CCN2 leads to less severe liver fibrosis compared with control mice exposed to high fat feeding and diabetes. Methods: Male control mice (B6.Cg-Tg(AlbCre)21Mgn/J (AlbCre) and homozygous CCN2fl/fl (CCN2fl/fl)), and hepatocyte specific CCN2 knockout mice (CCN2flxB6.Cg-Tg(AlbCre)21Mgn/J (CCN2KO)) were fed either standard chow or high fat diet (HFD; 45% kCal fat). HFD mice were rendered diabetic (HFD+DM) after 15 wks with low dose streptozotocin (2-3 x 0.65 mg/kg ip) then maintained for a further 10 wks. Liver fibrosis was assessed histologically with Picro Sirius Red (PSR) staining. Gene expression of fibrosis markers were measured by RT-qPCR. Biochemical assays were undertaken. Results: There was less fibrosis by PSR staining in the HFD+DM CCN2KO mice at the central vein (24% reduction, 2.20±0.84 controls vs. 1.66±0.82 CCN2-KO, p=0.03, one tailed Mann-Whitney test) and portal tract (18% reduction, 2.51±0.85 controls vs. 2.06±0.84 CCN2KO, p=0.04) compared with HFD+DM control mice. In HFD+DM CCN2KO mice, there were significant reductions in gene expression of collagen-I (44%, p=0.04), -III (44%, p=0.04) and -IVα1 (40%, p=0.01) but not in collagen-VI or fibronectin. Compared with chow fed mice, HFD+DM mice had higher body weight, BGLs, insulin levels, fat mass and AST and ALT activity but there was no difference in metabolic outcomes between control strains and CCN2KO mice. Conclusion: Our study has shown partial prevention of NASH fibrosis in diabetes in a novel model with hepatocyte specific CCN2 knockout mice. CCN2 is increasingly a potential target for limiting progression of NASH fibrosis in diabetes. Disclosure S. N. Parry: None. X. Wang: None. D. Min: None. J. S. Lourdesamy: None. P. F. Williams: None. A. Leask: Stock/Shareholder; Self; FibroGen. S. M. Twigg: Advisory Panel; Self; Abbott Diabetes, AstraZeneca, Boehringer Ingelheim Pharmaceuticals, Inc., Sanofi-Aventis, Consultant; Self; Aktivo Labs, Sanofi-Aventis, Research Support; Self; Abbott Diabetes, Speaker’s Bureau; Self; Abbott Diabetes, AstraZeneca.

Liquid Smoke as Fat Protector and Its Effect on Rumen Fermentation Characteristics and Microbial Activity

This study was conducted to determine the effect of liquid smoke as a fat protector on unsaturated fatty acids (UFAs) and its effect on rumen fermentation characteristics and microbial activity. Crude palm oil (CPO) was mixed with Prosteo skim milk (1:2), then divided into three treatments i.e., crude palm oil without protection by liquid smoke as a control (P0), crude palm oil protected by 2.5% of liquid smoke (P1), and crude falm oil protected by 5.0% of liquid smoke (P2). For in vitro testing, 300 mg of the feed substrate (elephant grass and bran with the ratio of 60:40) was added with 5% of each crude palm oil preparation of P0, P1, and P2 and put in a fermentor syringe. Then, 30 mL of the mixture of rumen fluid and buffer-minerals solution (1:2) was added into each syringe fermentor and flushed with CO2. The fermentor syringes were incubated in a water bath at 39ᵒC for 48 hours. Variables measured were fatty acid composition, fermentation characteristics, and rumen microbial activity. The data were analyzed by the analysis of variance with a completely randomized design. The results showed that the protection of CPO with liquid smoke in P1 and P2 groups decreased saturated fatty acids (SFAs), but increased (p<0.01) monounsaturated fatty acids (MUFAs), polyunsaturated fatty acids (PUFAs), and CMCase activity. Protection of CPO with 2.5% of liquid smoke (P1) significantly increased (p<0.01) fermentation characteristics (NH3 content and pH). It can be concluded that the use of 2.5% of liquid smoke has a better effect on feed fats protection, because it can reduce hydrogenation, increase UFAs, and has no negative effects on fermentation characteristics and microbial activity.

Does Consumption of Baker's Yeast (Saccharomyces cerevisiae) by Black Soldier Fly (Diptera: Stratiomyidae) Larvae Affect Their Fatty Acid Composition?

Fatty acids are important compounds for insects, but the requirements for essential fatty acids may differ between insect species. Most of the fatty acids are acquired through the insect’s diet; therefore, supplementing the diet with baker’s yeast (Saccharomyces cerevisiae Meyen ex E.C. Hansen), which produces unsaturated fatty acids, was predicted to affect the fatty acid composition of the insect. The tested insect was the black soldier fly (BSF) (Hermetia illucens L.), that is used as a source of protein and fat in feed. Therefore, there is importance for BSF larvae (BSFL) nutritional composition, especially the unsaturated fatty acids content, which is one of the nutritional limitations for mammalian diets. The dominant fatty acids of the tested BSFL were the saturated fatty acids: lauric, myristic, and palmitic acids, as found in other BSF studies. Oleic acid (c18:1) and linoleic acid (C18:2) were the abundant unsaturated fatty acids in the BSFL. The proportion of linoleic acid was higher in the substrate with the supplemental yeast; however, this did not affect its proportion in the larvae. The higher proportion of linoleic acid may have been exploited as a source for production of saturated lauric acid. Therefore, providing unsaturated fatty acids to the substrate through supplemental baker’s yeast is not the most efficient way to increase the proportion of unsaturated fatty acids in the larvae.

Low‐dose lithium supplementation augments mitochondrial biogenesis in murine skeletal muscle after high fat feeding

Promotion of oxidative metabolism in skeletal muscle improves muscle performance, fatigue resistance, and may help combat the obesogenic effects of a high fat diet. Lithium is a well-known glycogen synthase kinase (GSK3) inhibitor and we have recently shown that low-dose lithium supplementation (10 mg kg-1 day-1) augments skeletal muscle oxidative capacity in mice. In the soleus muscle, lithium mediated GSK3 inhibition increased the expression of oxidative markers such as peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC-1a) and myosin heavy chain I (MHCI) leading to improvements in muscle contractility and fatigue resistance. Interestingly, this same dose of lithium (10 mg kg-1 day-1) has been previously shown to attenuate high fat diet induced weight gain and metabolic efficiency (weight gained per calories consumed). However, the impact of lithium supplementation on muscle oxidative phenotype after high fat feeding has yet to be investigated. In this study, 36 wild-type male C57BL/6J mice were randomly assigned to low-fat standard chow diet (LFD, 4% kcal fat), high-fat diet (HFD, 60% kcal fat), or HFD with lithium chloride (HFD + Li; 10 mg kg-1 day-1) treatment groups for 12 weeks. Soleus muscles were collected at the end for Western blot analysis of total and phosphorylated GSK3 (inhibitory sites serine9 for GSK3b and serine21 for GSK3a), PGC-1a, cytochrome c oxidase subunit IV (COXIV), and MHCI protein content. Our results showed that the level of phosphorylated GSK3a and GSK3b (relative to total GSK3) were significantly lower after the HFD (-41 and -44%, respectively, p < 0.05), but in the HFD + Li group levels were similar to the LFD group. Therefore, the HFD enhanced soleus muscle GSK3 activation, and lithium supplementation was able to reduce this effect. No significant changes in MHCI content were observed. When examining PGC-1a, we found that the HFD + Li group had protein levels that were nearly 2-fold higher vs LFD, albeit only trending towards significance (p = 0.06). Further, COXIV content progressively increased from LFD, to HFD, and to HFD + Li (linear trend, r2=0.26, p = 0.002) with both HFD and HFD + Li groups having significantly higher COXIV content when compared to LFD. Altogether, our data shows that GSK3 inhibition via low dose lithium supplementation can augment mitochondrial biogenesis in the presence of a HFD. Future studies will further examine whether GSK3 inhibition via lithium can augment muscle oxidative metabolism after high fat feeding.

Neuronatin Promotes SERCA Uncoupling and Its Expression is Inversely Associated with High Fat Diet Induced Weight Gain in vivo

Neuronatin (NNAT) is a paternally imprinted gene involved in many aspects of metabolism but the underlying cellular mechanism remains unknown. Its sequence homology with phospholamban (PLN) and sarcolipin (SLN) suggest that NNAT has a putative role in regulating the enzymatic activity of sarco(endo)plasmic reticulum Ca2+-ATPase (SERCA) in striated muscles. Both PLN and SLN physically interact and functionally inhibit SERCA by lowering its affinity for Ca2+, but only SLN has the unique ability to uncouple SERCA-mediated Ca2+ transport from ATP hydrolysis in mouse soleus muscles. Uncoupling the SERCA pump increases energy expenditure and may be beneficial in combatting energy imbalances, such as obesity. The objective of this study was to test our hypothesis that like SLN, NNAT would also promote SERCA uncoupling. In addition, we also examined whether NNAT expression would be altered in soleus muscles obtained from C57BL/6 mice fed a high fat diet (HFD). Using human embryonic kidney (HEK) cells co-transfected with SERCA and NNAT cDNA, we measured SERCA coupling ratio using an Indo-1 based fluorometric Ca2+ uptake assay and an enzyme-linked spectrophotometric ATPase assay. NNAT promoted uncoupling of both SERCA1a and SERCA2a isoforms in HEK cells in a dose-dependent manner with SERCA1a showing increased sensitivity when compared to SERCA2a. Western blotting was used to confirm HEK cell transfection as well as to assess NNAT and SERCA content in soleus muscles from mice fed a low (LFD) or HFD (60% kcal) for 12 weeks. A ~40% reduction in SERCA1a and a ~50% increase in SERCA2a protein content was observed in the soleus muscles of mice fed a HFD compared to LFD. Total NNAT content and its levels relative to SERCA (SERCA1a + SERCA2a) were reduced by ~40% and ~45%, respectively in HFD mice compared to LFD mice. Plotting muscle NNAT content (relative to SERCA) against total weight gained after 12 weeks of high fat feeding also revealed a significant inverse relationship (r2=0.49, p=0.01), suggesting that higher NNAT content (relative to SERCA) is related to reduced weight gained from a HFD in vivo. This is the first study to demonstrate NNAT as a SERCA uncoupler and that its expression in muscle is negatively associated with HFD induced weight gain. These findings suggest that NNAT may regulate whole-body metabolism and energy balance via SERCA uncoupling.

Exploring the role of ATP10A in diet‐induced obesity, insulin resistance, and type 2 diabetes

Type IV P-type ATPases (P4-ATPases) are enzymes that catalyze the translocation of lipids across plasma membranes. Our lab recently discovered that Atp10a, a mammalian P4-ATPase, can translocate glucosylceramide, a sphingolipid. Both Atp10a and glucosylceramide have been independently implicated in metabolic dysfunction. To explore the role of Atp10a in metabolic dysfunction and sphingolipid metabolism, we created a novel Atp10a knockout (KO) mouse model. Atp10a KO mice display a female-specific excess weight gain on standard chow that is exacerbated by 12 weeks of high fat feeding and this is attributable to increased adiposity. Additionally, they display an accumulation of neutral lipids and altered ceramide metabolism in their livers. After a 5 hour fast, these mice exhibit elevated blood glucose levels that are not coupled to a proportional elevation in insulin levels. Atp10a KO mice also exhibit elevated plasma free fatty acids, cholesterol, and triglycerides. Thus far, our studies have shown that knocking out Atp10a in a mouse results in sex-specific perturbations to body weight and composition, glucose homeostasis, plasma lipid levels, and liver metabolism.

Adult rats from undernourished dams show sex-dependent impaired expression in taste papillae and hypothalamus of genes responsible for sweet and fat detection and signalling

ABSTRACT Aim Individuals undernourished in utero or during early life are at high risk of developing obesity and metabolic disorders and show an increased preference for consuming sugary and fatty food. This study aimed at determining whether impaired taste detection and signalling in the lingual epithelium and the brain might contribute to this altered pattern of food intake. Methods The preference for feeding fat and sweet food and the expression in circumvallate papillae and hypothalamus of genes coding for sweet and fat receptors and transducing pathways were evaluated in adult rats born to control or calorie-restricted dams. Expression in the hypothalamus and the brain's reward system of genes involved in the homeostatic and hedonic control of food intake was also determined. Results Male and female undernourished animals exhibited increased expression in taste papillae and hypothalamus of T1R1, T1R2, CD36, gustducin, TRMP5 and PLC-β2 genes, all of which modulate sweet and fat detection and intracellular signalling. However, the severity of the effect was greater in females than in males. Moreover, male, but not female, undernourished rats consumed more standard and sweetened food than their control counterparts and presented increased hypothalamic AgRP and NPY mRNAs levels together with enhanced dopamine transporter and dopamine receptor D2 expression in the ventral tegmental area. Conclusions Maternal undernutrition induces sex-specific changes in food preferences and gene expression in taste papillae, hypothalamus and brain reward regions. The gene expression alterations in the male offspring are in line with their preference for consuming sugary and fatty food.

Ethanol Extract Combinations Effect of Celery Herb (&lt;i&gt;Apium graveolens&lt;/i&gt; L.) and Bay Leaf (&lt;i&gt;Syzygium polyanthum &lt;/i&gt;W.) Toward Hypertensive Mice Induced by Sodium Chloride and High Fat Feed

The combination of herb medicine is alternative option in hypertension because it has more potential for treatment with complications like hyperlipida. Plant which can be used for anti-hypertension therapy is combination of celery herb (Apium graveolens) and bay leaf ethanol extract (Syzygium polyanthum). This research aims to determine the activity of a combination of celery herb ethanol extract (CHEE) and bay leaf ethanol extract (BLEE) and find out how much the decrease of blood pressure on the combination of both toward hypertension with high fat Wistar mice. The design of this research used an experimental design with pre-post control group design. Hypertensive mice are induced with high fat feed and orally with sodium chloride 8%, then the mice are supplied with combinations extract with the dose of 1.125 ; 6.25 ; 2.25 ; 12.5 and 4.5 ; 25 mg/kg, hydrochlorotiazide 2.25 mg/kg, Simvastatin 0.9 mg/kg, CHEE 4.5 mg/kg, BLEE 25 mg/kg and CMC-Na 0.5%. Research result shows that the combination can decrease systole blood pressure in the 22nd day. The extract combination has anti-hypertension effect (it is able to decrease systole blood pressure ≥ 20 mmHg) and it is not significantly different with normal group (p&lt;0.05). Based on the research, it can be concluded that ethanol extract combination can decrease systole blood pressure with high fat complications after using it for 22 days.

Test Activity Combinations of Celery Herb (&lt;i&gt;Apium graveolens&lt;/i&gt; L.) and Bay Leaf (&lt;i&gt;Syzygium polyanthum&lt;/i&gt; W.) Ethanol Extract Toward DecreasedLipid Profile Levels in Hypertensive Mice

Celery herb contains flavonoid which can hamper cholesterol synthesis through inhibitor HMG-CoA reductase. Bay leaf has a working mechanism that stimulates bile fluid secretion and stimulates blood circulation. This research aims to know the activity combinations of bay leaf ethanol extract (BLEE) and celery herb ethanol extract (CHEE) and to know which dose mostly can decrease lipid profile levels. The design of this research uses experimental withpre-post control group design.The mice are rendered with NaCl 8% solution and High Fat Feed. The mice are divided into 9 groups, in each group there are 5 mice which are normal group, control group, HCT group, Simvastatin group, and CHEE 4.50 mg/kgBW, BLEE 25.00 mg/kgBW, combination 1, combination 2, and combination 3. Data analysis is conducted with Normality test,One Way ANOVAtest,Kruskal-Wallis,Mann-Whitney. Based on the research after they are induced with NaCl 8% solvent and given high-fat feed, the extract can increase total cholesterol levels, triglycerides, LDL, and also significantly decrease HDL (p &lt;0.05). After giving a test extract solution, it can reduce total cholesterol, triglycerides, LDL, and also significantly increase HDL levels (p &lt;0.05). The conclusion, the three variations combinations dose can decrease total cholesterol, Triglycerides, LDL, and also increase HDL levels, and combination dose 3 has the greatest decrease in total cholesterol, triglyceride, LDL, and the highest increase in HDL levels. So the lower the total cholesterol level, the lower the possibility of hypertension.

МОРФО-БИОХИМИЧЕСКИЙ СТАТУС МОЛОДНЯКА СВИНЕЙ НА ОТКОРМЕ ПРИ ИСПОЛЬЗОВАНИИ В ИХ РАЦИОНАХ КОРМОВЫХ ДОБАВОК «ПОЛИСОЛ ОМЕГА-3» И «ОМЕГА-3 АКТИВ»

The article describes results of application of «Polysol Omega-3» and «Omega-3 Active» feed additives in rations of young pigs during fattening period and their influence on hematological and biochemical parameters of animals‘ blood. The research was carried out at the pig breeding complex SPK named after N.K. Krupskaya of Melekesskyi district of Ulyanovsk region. The basis of «Omega-3 Active» supplement is flaxseed oil, which contains 60% of polyunsaturated fatty acids and the prevailing proportion of Omega-3. The feed mixture «Polysol Omega-3» includes fatty acids in combination with a complex of biologically active and mineral substances. In addition to studying the productivity and quality of pig production, the study of the effect of these additives on morpho- biochemical status of blood and, in particular, lipid metabolism is of scientific interest. The results of the analysis showed that in case of application of these feed additives in the rations of young pigs, the concentration of erythrocytes in their blood was higher in comparison with control analogs by 1.13 ... 3.56 %, hemoglobin by 7.97 ... 14.15 % (P &lt;0.01), the total protein content by 6 , 75 ... 7.88 %. This is confirmed by improvement of redox processes and metabolism in their bodies. The ration enrichment with the analyzed additives contributed to better usage of fats in feed, which is confirmed by the studied parametres of lipid metabolism (cholesterol, HDL, LDL, triglycerides). The activation of biosynthetic processes had a positive effect on average daily gain of pigs, as a result of which the early maturity of animals increased by 9 ... 23 days

High fat induces activation of the tryptophan-ERK-CREB pathway and promotes bone absorption in cage layers

Cage layer fatigue is a common metabolic disease associated with a calcium and phosphorus imbalance, but recently we found this disease can be led by high fat diet. In order to elucidate the pathogenesis induced by a high fat diet, we randomly divided 88 White Shell Roman layers into 2 groups. There were 44 layers in each group. The control group was fed by a standard layer rations, and the high fat group was fed by completed rations mixing with 3% soybean oil. This study successfully constructed an animal model of osteoporosis caused by high fat. Bone samples were collected for bone mineral density, bone biomechanical properties which are all decreased at 26, 30, 34, and 38 wk old. We found the pathway of tryptophan-ERK-CREB from the perspective of metabonomics which promote the bone absorption. By metabolomics, we screened the significantly activated tryptophan pathway in high fat feed and detected the elevated tryptophan metabolite serum 5-HT at 26, 30, 34 and 38 wk old in the high fat group. At 38 wk old, we detected significantly elevated protein and mRNA levels of ERK/CREB/C-fos in bone tissue in the high fat group. So we concluded that high-fat were associated with a decrease in bone density and bone biomechanical index by disrupting tryptophan-5-HT-ERK1/2-CREB metabolism signaling pathways.

Peptidomics of enteroendocrine cells and characterisation of potential effects of a novel preprogastrin derived-peptide on glucose tolerance in lean mice

ObjectivesTo analyse the peptidomics of mouse enteroendocrine cells (EECs) and human gastrointestinal (GI) tissue and identify novel gut derived peptides. MethodsHigh resolution nano-flow liquid chromatography mass spectrometry (LC–MS/MS) was performed on (i) flow-cytometry purified NeuroD1 positive cells from mouse and homogenised human intestinal biopsies, (ii) supernatants from primary murine intestinal cultures, (iii) intestinal homogenates from mice fed high fat diet. Candidate bioactive peptides were selected on the basis of species conservation, high expression/biosynthesis in EECs and evidence of regulated secretionin vitro. Candidate novel gut-derived peptides were chronically administered to mice to assess effects on food intake and glucose tolerance. ResultsA large number of peptide fragments were identified from human and mouse, including known full-length gut hormones and enzymatic degradation products. EEC-specific peptides were largely from vesicular proteins, particularly prohormones, granins and processing enzymes, of which several exhibited regulated secretion in vitro. No regulated peptides were identified from previously unknown genes. High fat feeding particularly affected the distal colon, resulting in reduced peptide levels from GCG, PYY and INSL5. Of the two candidate novel peptides tested in vivo, a peptide from Chromogranin A (ChgA 435−462a) had no measurable effect, but a progastrin-derived peptide (Gast p59−79), modestly improved glucose tolerance in lean mice. ConclusionLC–MS/MS peptidomic analysis of murine EECs and human GI tissue identified the spectrum of peptides produced by EECs, including a potential novel gut hormone, Gast p59−79, with minor effects on glucose tolerance.

Restoring Perivascular Adipose Tissue Function in Obesity Using Exercise

PurposePerivascular adipose tissue (PVAT) exerts an anti-contractile effect which is vital in regulating vascular tone. This effect is mediated via sympathetic nervous stimulation of PVAT by a mechanism which involves noradrenaline uptake through organic cation transporter 3 (OCT3) and β3-adrenoceptor-mediated adiponectin release. In obesity, autonomic dysfunction occurs, which may result in a loss of PVAT function and subsequent vascular disease. Accordingly, we have investigated abnormalities in obese PVAT, and the potential for exercise in restoring function.MethodsVascular contractility to electrical field stimulation (EFS) was assessed ex vivo in the presence of pharmacological tools in ±PVAT vessels from obese and exercised obese mice. Immunohistochemistry was used to detect changes in expression of β3-adrenoceptors, OCT3 and tumour necrosis factor-α (TNFα) in PVAT.ResultsHigh fat feeding induced hypertension, hyperglycaemia, and hyperinsulinaemia, which was reversed using exercise, independent of weight loss. Obesity induced a loss of the PVAT anti-contractile effect, which could not be restored via β3-adrenoceptor activation. Moreover, adiponectin no longer exerts vasodilation. Additionally, exercise reversed PVAT dysfunction in obesity by reducing inflammation of PVAT and increasing β3-adrenoceptor and OCT3 expression, which were downregulated in obesity. Furthermore, the vasodilator effects of adiponectin were restored.ConclusionLoss of neutrally mediated PVAT anti-contractile function in obesity will contribute to the development of hypertension and type II diabetes. Exercise training will restore function and treat the vascular complications of obesity.

Impact of obesity on day-night differences in cardiac metabolism.

An intrinsic property of the heart is an ability to rapidly and coordinately adjust flux through metabolic pathways in response to physiologic stimuli (termed metabolic flexibility). Cardiac metabolism also fluctuates across the 24‐hours day, in association with diurnal sleep‐wake and fasting‐feeding cycles. Although loss of metabolic flexibility has been proposed to play a causal role in the pathogenesis of cardiac disease, it is currently unknown whether day‐night variations in cardiac metabolism are altered during disease states. Here, we tested the hypothesis that diet‐induced obesity disrupts cardiac “diurnal metabolic flexibility”, which is normalized by time‐of‐day‐restricted feeding. Chronic high fat feeding (20‐wk)‐induced obesity in mice, abolished diurnal rhythms in whole body metabolic flexibility, and increased markers of adverse cardiac remodeling (hypertrophy, fibrosis, and steatosis). RNAseq analysis revealed that 24‐hours rhythms in the cardiac transcriptome were dramatically altered during obesity; only 22% of rhythmic transcripts in control hearts were unaffected by obesity. However, day‐night differences in cardiac substrate oxidation were essentially identical in control and high fat fed mice. In contrast, day‐night differences in both cardiac triglyceride synthesis and lipidome were abolished during obesity. Next, a subset of obese mice (induced by 18‐wks ad libitum high fat feeding) were allowed access to the high fat diet only during the 12‐hours dark (active) phase, for a 2‐wk period. Dark phase restricted feeding partially restored whole body metabolic flexibility, as well as day‐night differences in cardiac triglyceride synthesis and lipidome. Moreover, this intervention partially reversed adverse cardiac remodeling in obese mice. Collectively, these studies reveal diurnal metabolic inflexibility of the heart during obesity specifically for nonoxidative lipid metabolism (but not for substrate oxidation), and that restricting food intake to the active period partially reverses obesity‐induced cardiac lipid metabolism abnormalities and adverse remodeling of the heart.

Hyperinsulinemia Induced Altered Insulin Signaling Pathway in Muscle of High Fat- and Carbohydrate-Fed Rats: Effect of Exercise.

Insulin resistance is a state of impaired responsiveness to insulin action. This condition not only results in deficient glucose uptake but increases the risk for cardiovascular diseases (CVD), stroke, and obesity. The present work investigates the molecular mechanisms of high carbohydrate and fat diet in inducing prediabetic hyperinsulinemia and effect of exercise on InsR signaling events, muscular AChE, and lactate dehydrogenase activity. Adult male Wistar rats were divided into the control (C) diet group, high-carbohydrate diet (HCD) group, high-fat diet (HFD) group, and HCD and HFD groups with exercise (HCD Ex and HFD Ex, respectively). Acetyl choline esterase activity, lactate dehydrogenase activity, total lactate levels, IRS1 phosphorylations, and Glut4 expression patterns were studied in the muscle tissue among these groups. High carbohydrate and fat feeding led to hyperinsulinemic status with reduced acetylcholine esterase (AChE) activity and impaired phosphorylation of IRS1 along with increased lactate concentrations in the muscle. Exercise significantly upregulated phosphoinositide 3 kinase (PI3K) docking site phosphorylation and downregulated the negative IRS1 phosphorylations thereby increasing the glucose transporter (GLUT) expressions and reducing the lactate accumulation. Also, the levels of second messengers like IP3 and cAMP were increased with exercise. Increased second messenger levels induce calcium release thereby activating the downstream pathway promoting the translocation of GLUT4 to the plasma membrane. Our results showed that the metabolic and signaling pathway dysregulations seen during diet-induced hyperinsulinemia, a metabolic condition seen during the early stages in the development of prediabetes, were improved with vigorous physical exercise. Thus, exercise can be considered as an excellent management approach over drug therapy for diabetes and its complications.