Standard Laboratory Chow Research Articles

Abstract P138: Diet Influences the Expression of Renal Sodium-Glucose Cotransporters in Juvenile Male Rats

Poor nutrition and metabolic dysfunction are becoming more prevalent among children, as evident by increasing childhood obesity in recent years. The kidney sodium-glucose cotransporter 2 (SGLT2) is a contributor to overall metabolic homeostasis, with inhibition of SGLT2 significantly reducing circulating glucose and lipids. The study objective was to determine if diet influences the expression of renal glucose transporters in young rats. Weanling male Wistar Kyoto rats received 1) standard laboratory chow, 2) AIN-93G diet, or 3) a high-fat diet (60% kcal, HFD). Each dietary group was then further subdivided to receive either filtered water or 5% (w/v) sucrose solution ad libitum . Four weeks later, animals were euthanized and kidneys were collected and processed. mRNA expression of Sglt1 , Sglt2 , Sqstm1 , and Nlrp3 was determined using gene specific primers and RT-PCR. Renal SGLT2 protein expression was visualized using immunohistochemistry and quantified using Imagej software. Diet significantly influenced the renal expression of both Sglt1 ( P =0.005) and Sglt2 (P=0.046), while beverage did not have a significant effect on either ( P =0.7 and P =0.4, respectively). Rats fed the HFD had more than 30% reduction in Sglt1 mRNA expression as compared to rats fed chow, and ~20% reduction in Sglt2 as compared to rats fed AIN. Preliminary data suggest that diet and beverage significantly altered renal protein expression of SGLT2, with rats fed chow having the lowest renal expression that was slightly increased with sugar consumption (1.5% versus 2.7%, respectively). Rats fed AIN had the greatest expression, but was drastically reduced in rats consuming sugar (4% versus 1.2%, respectively). To determine if autophagy and inflammasome activation is involved, the expression of Sqstm1 and Nlrp3 was measured by RT-PCR. Similarly, rats fed HFD had the lowest expression of both Sqstm1 ( P<0.001) and Nlrp3 ( P=0.03). Sqstm1 and Nlrp3 mRNA expression was decreased with sugar consumption in rats fed chow and AIN, whereas Sqstm1 ( P =0.01) and Nlrp3 ( P <0.001) mRNA expression was increased in rats fed HFD with sugar beverage. Our data suggest that diet significantly influences renal glucose transporter expression in juvenile rats in a way that may be mediated by autophagic impairment. The modulatory effect of sugar consumption on SGLT2 protein, Sqstm1, and Nlrp3 was diet-dependent, highlighting the importance of nutrition to renal physiology within weeks after weaning.

SCD4 deficiency decreases cardiac steatosis and prevents cardiac remodeling in mice fed a high-fat diet

Stearoyl-CoA desaturase (SCD) is a lipogenic enzyme that catalyzes formation of the first double bond in the carbon chain of saturated fatty acids. Four isoforms of SCD have been identified in mice, the most poorly characterized of which is SCD4, which is cardiac-specific. In the present study, we investigated the role of SCD4 in systemic and cardiac metabolism. We used WT and global SCD4 KO mice that were fed standard laboratory chow or a high-fat diet (HFD). SCD4 deficiency reduced body adiposity and decreased hyperinsulinemia and hypercholesterolemia in HFD-fed mice. The loss of SCD4 preserved heart morphology in the HFD condition. Lipid accumulation decreased in the myocardium in SCD4-deficient mice and in HL-1 cardiomyocytes with knocked out Scd4 expression. This was associated with an increase in the rate of lipolysis and, more specifically, adipose triglyceride lipase (ATGL) activity. Possible mechanisms of ATGL activation by SCD4 deficiency include lower protein levels of the ATGL inhibitor G0/G1 switch protein 2 and greater activation by protein kinase A under lipid overload conditions. Moreover, we observed higher intracellular Ca2+ levels in HL-1 cells with silenced Scd4 expression. This may explain the activation of protein kinase A in response to higher Ca2+ levels. Additionally, the loss of SCD4 inhibited mitochondrial enlargement, NADH overactivation, and reactive oxygen species overproduction in the heart in HFD-fed mice. In conclusion, SCD4 deficiency activated lipolysis, resulting in a reduction of cardiac steatosis, prevented the induction of left ventricular hypertrophy, and reduced reactive oxygen species levels in the heart in HFD-fed mice.

Mesenchymal Stem Cells Suppress Kidney Injury molecule-1 Associated with Inhibition of Renal PKC/ NF-Kβ / STAT3 Fibrotic Signaling Pathway in Rats with Diabetic Nephropathy

IntroductionBackground: Diabetes stands as the predominant etiology behind end-stage kidney disease, commonly referred to as renal failure. The intricate interplay among oxidative stress, inflammation, and renal fibrotic changes in diabetes-induced nephropathy, particularly in instances involving and not involving the administration of mesenchymal stem cells (MSCs), remains a subject less explored in existing research.Material and methodsMethods: Twenty-four male Wistar rats (180 and 200 grams) were randomly assigned to one of three groups (n = 8). The control group received standard laboratory chow, and the groups with T2DM received a single dose of streptozotocin, 45 mg/kg, after three weeks of pretreatment with a high-fat diet (HFD). Rats with T2DM were split into the T2DM model group and Bone marrow (BM) mesenchymal stem cells (MSCs) treated group (T2DM+MSCs) eight weeks after DM was confirmed. BM-MSCs were injected systemically at 2 × 106 cells/rat doses.ResultsResults: Diabetes significantly altered oxidative stress (MDA, SOD), inflammation (TNFα, IL-6), and kidney injury (KIM-1, NAGAL) biomarkers, a modulation that was mitigated by MSCs (p < 0.0001). Furthermore, diabetes-induced kidney fibrosis showed a noteworthy reduction in the presence of MSCs. A notable correlation emerged between body weight, systolic blood pressure (SBP), oxidative stress, inflammation, fibrosis, the PKC/NF-KB/STAT-3 axis, and hyperglycemia.ConclusionsConclusions: Our results suggest that diabetes was associated with elevated oxidative stress, inflammation, biomarkers of kidney injury, upregulation of the renal PKC/NF-KB/STAT-3 pathway, and hypertension, all countered by MSCs intervention.

The Effects of Eating a Traditional High Fat/High Carbohydrate or a Ketogenic Diet on Sensitivity of Female Rats to Morphine.

Patients diagnosed with obesity are prescribed opioid medications at a higher rate than the general population; however, it is not known if eating a high fat diet might impact individual sensitivity to these medications. To explore the hypothesis that eating a high fat diet increases sensitivity of rats to the effects of morphine, 24 female Sprague-Dawley rats (n = 8/diet) ate either a standard (low fat) laboratory chow (17% kcal from fat), a high fat/low carbohydrate (ketogenic) chow (90.5% kcal from fat), or a traditional high fat/high carbohydrate chow (60% kcal from fat). Morphine-induced antinociception was assessed using a warm water tail withdrawal procedure, during which latency (in seconds) for rats to remove their tail from warm water baths was recorded following saline or morphine (0.32-56 mg/kg, i.p.) injections. Morphine was administered acutely and chronically (involving 18 days of twice-daily injections, increasing in 1/4 log dose increments every 3 days: 3.2-56 mg/kg, i.p., to induce dependence and assess tolerance). The adverse effects of morphine (i.e., tolerance, withdrawal, and changes in body temperature) were assessed throughout the study. Acute morphine induced comparable antinociception in rats eating different diets, and all rats developed tolerance following chronic morphine exposure. Observable withdrawal signs and body temperature were also comparable among rats eating different diets; however, withdrawal-induced weight loss was less severe for rats eating ketogenic chow. These results suggest that dietary manipulation might modulate the severity of withdrawal-related weight loss in ways that could be relevant for patients.

Dietary Cinnamaldehyde Activation of TRPA1 Antagonizes High-Salt-Induced Hypertension Through Restoring Renal Tubular Mitochondrial Dysfunction.

The renal proximal tubule (RPT) plays a pivotal role in regulating sodium reabsorption and thus blood pressure (BP). Transient receptor potential ankyrin 1 (TRPA1) has been reported to protect against renal injury by modulating mitochondrial function. We hypothesize that the activation of TRPA1 by its agonist cinnamaldehyde may mitigate high-salt intake-induced hypertension by inhibiting urinary sodium reabsorption through restoration of renal tubular epithelial mitochondrial function. Trpa1-deficient (Trpa1-/-) mice and wild-type (WT) mice were fed standard laboratory chow [normal diet (ND) group, 0.4% salt], standard laboratory chow with 8% salt [high-salt diet (HS) group], or standard laboratory chow with 8% salt plus 0.015% cinnamaldehyde [high-salt plus cinnamaldehyde diet (HSC) group] for 6 months. Urinary sodium excretion, reactive oxygen species (ROS) production, mitochondrial function, and the expression of sodium hydrogen exchanger isoform 3 (NHE3) and Na+/K+-ATPase of RPTs were determined. Chronic dietary cinnamaldehyde supplementation reduced tail systolic BP and 24-hour ambulatory arterial pressure in HS-fed WT mice. Compared with the mice fed HS, cinnamaldehyde supplementation significantly increased urinary sodium excretion, inhibited excess ROS production, and alleviated mitochondrial dysfunction of RPTs in WT mice. However, these effects of cinnamaldehyde were absent in Trpa1-/- mice. Furthermore, chronic dietary cinnamaldehyde supplementation blunted HS-induced upregulation of NHE3 and Na+/K+-ATPase in WT mice but not Trpa1-/- mice. The present study demonstrated that chronic activation of Trpa1 attenuates HS-induced hypertension by inhibiting urinary sodium reabsorption through restoring renal tubular epithelial mitochondrial function. Renal TRPA1 may be a potential target for the management of excessive dietary salt intake-associated hypertension.

Histological and biochemical effects of an ethanolic extract of Myrtus communis leaf on the pancreases of rats fed high fat diets

ABSTRACT We investigated the effects of an ethanolic extract of Myrtus communis subsp. communis (MC) leaves on the pancreases of rats fed with a high fat diet (HFD). Wistar albino rats were fed either with standard lab chow (Control group) or with a 45% fat diet (HFD and HFD+MC groups) for 4 months, with the MC extract (100 mg/kg) being administered by orogastric gavage to rats in the HFD+MC group during the last month. Blood and pancreas samples were collected from all experimental groups at the end of the study. Insulin and leptin levels, and the lipid profile, were analyzed in the blood serum. Pancreatic injury was assessed histologically. Insulin, nuclear factor kappa beta (NF-κB), and alpha-smooth muscle actin (α-SMA) were assessed using immunohistochemistry. Apoptosis was assessed using terminal deoxynucleotidyl transferase dUTP nick end-labeling (TUNEL) immunohistochemistry. In addition, oxidant/antioxidant activity was analyzed by biochemical methods. Increased body weight, serum insulin and leptin levels, blood glucose level and pancreatic tissue malondialdehyde (MDA), 8-hydroxy-2’-deoxyguanosine (8-OHdG) levels, myeloperoxidase (MPO) activity and decreased tissue glutathione (GSH) level were observed in the HFD group compared to the Control group, in addition to dyslipidemia. An increased histopathological damage score, pancreatic islet area, insulin, TUNEL, NF-κB and α-SMA immunoreactivity were seen in animals from the HFD group compared to the Control group. However, such pathological changes were reduced in the HFD+MC group. Our data indicate further investigation of MC extract as a therapeutic adjuvant for HFD-induced pancreatic injury, acting via anti-inflammatory and antioxidant mechanisms, is worth carrying out.

Mesenchymal stem cells improve cardiac function in diabetic rats by reducing cardiac injury biomarkers and downregulating JAK/STAT/iNOS and iNOS/Apoptosis signaling pathways

Cardiovascular complications are prevalent manifestations of type 2 diabetes mellitus (T2DM) and are usually the main cause of death. This study aims to show the underlying mechanisms of the potential therapeutic effect of mesenchymal stem cells (MSCs) on diabetic cardiac dysfunction. Twenty-four male Wistar rats were randomly assigned to one of three groups The control group received standard laboratory chow, and the groups with T2DM received a single dose of 45 mg/kg body weight of streptozotocin (STZ) after 3 weeks of pretreatment with a high-fat diet (HFD). Eight weeks after the diagnosis of T2DM, rats were divided into two groups: the T2DM model group and the T2DM + MSCs group. BM-MSCs were administered systemically at 2 × 106 cells/rat doses.A Significant amelioration in Homeostatic Model Assessment of Insulin Resistance (HOMA-IR) and dyslipidemia was noted 2 weeks post-administration of MSCs. Administration of MSCs improved dyslipidemia, the altered cardiac injury biomarkers (p ≤ 0.0001), downregulated Janus kinase 2/signal transducer and activator of transcription 3(JAK2/STAT3)/inducible Nitric oxide synthase (iNOS) and iNOS/Apoptosis signaling pathways. This was associated with improved cardiac dysfunction (impaired left ventricular performance and decreased contractility index).Our results show that MSCs ameliorate cardiac dysfunction associated with diabetic cardiomyopathy by lowering dyslipidemia and insulin resistance, inhibiting oxidative stress, and inflammation, downregulating JAK2/STAT3/iNOS and iNOS/Apoptosis signaling pathways.

Effect of high-fat diet on isometric, concentric and eccentric contractile performance of skeletal muscle isolated from female CD-1 mice.

Despite evidence inferring muscle and contractile mode-specific effects of high-fat diet (HFD), no study has yet considered the impact of HFD directly on eccentric muscle function. The present work uniquely examined the effect of 20-week HFD on the isometric, concentric and eccentric muscle function of isolated mouse soleus (SOL) and extensor digitorum longus (EDL) muscles. CD-1 female mice were randomly split into a control (n=16) or HFD (n=17) group and for 20 weeks consumed standard lab chow or HFD. Following this period, SOL and EDL muscles were isolated and assessments of maximal isometric force and concentric work loop (WL) power were performed. Each muscle was then subjected to either multiple concentric or eccentric WL activations. Post-fatigue recovery, as an indicator of incurred damage, was measured via assessment of concentric WL power. In the EDL, absolute concentric power and concentric power normalised to muscle mass were reduced in the HFD group (P<0.038). HFD resulted in faster concentric fatigue and reduced eccentric activity-induced muscle damage (P<0.05). For the SOL, maximal isometric force was increased, and maximal eccentric power normalised to muscle mass and concentric fatigue were reduced in the HFD group (P<0.05). HFD effects on eccentric muscle function are muscle-specific and have little relationship with changes in isometric or concentric function. HFD has the potential to negatively affect the intrinsic concentric and eccentric power-producing capacity of skeletal muscle, but a lack of a within-muscle uniform response indicates disparate mechanisms of action which require further investigation.

Abstract P478: Short-Term Voluntary Exercise Significantly Influences the Diet-Induced Changes in Cecal Microbial Content in Male Mice

Two leading environmental factors that influence the overall health of humans are diet and physical exercise. However, it is still unclear how these factors influence health and disease susceptibility. Both diet and exercise have been shown to modulate the microbial flora that may significantly influence health and disease; however, it is unclear if diet modulates the effects of exercise on gut microflora. Therefore, this study aimed to determine how diet and exercise modulate the microbial content of the mouse cecum. Male, weanling, C57Bl/6 mice were randomly assigned to receive one of three diets ad libitum: standard laboratory chow, AIN93-G, and our novel Americanized diet (AD). Within each diet, mice were then assigned to a voluntary exercise regimen (wheel access, 16-hours per day, 3-days per week) or a control sedentary group (individually housed in cage without wheel for the same duration and frequency). After 2-weeks of exercise, mice were euthanized and cecal microbial content was determined by16S rDNA sequencing and RT-PCR for Firmicutes (FIRM), Bifidobacterium (GBIFD), Lactobacillus (LB) , and Bacteroides (BACT). Data were analyzed using GLM Procedures in SPSS with significance determined at P&lt;0.05. Pilot results found that diet, exercise, and their interaction significantly influenced FIRM expression. Mice fed the standard chow diet had the lowest expression of FIRM , which was significantly lower than mice fed AIN (P&lt;0.001) and AD (P=0.001). Exercise caused a significant decrease (P&lt;0.001) in FIRM presence, especially in mice fed AIN and AD. Diet and exercise also significantly influenced GBIFD , with mice fed AIN having significantly more GBIFD than mice fed chow and AD (P&lt;0.001 for both). Exercise also caused a significant reduction (P=0.016) in GBIFD , and a reduction in LB expression in mice fed chow and AD, but not AIN (P=0.03). The results of this study highlight how two key aspects of lifestyle, diet and exercise, independently and additively influence the gastrointestinal flora in a preclinical mouse model. Additional studies are needed to validate these findings and determine their physiological significance.

Abstract P485: Exercise Influences Metabolic Parameters and Expression of Markers of Endoplasmic Reticulum Stress in a Diet-Dependent Manner

The relationship between diet, exercise, and metabolic dysfunction is widely established; however the mechanisms responsible remain unclear. Endoplasmic reticulum stress (ERS) has been shown to play a significant role in metabolic homeostasis and disease processes; however it is unclear how diet and exercise interact to influence ERS. Therefore, the objective of this project was to determine how diet and exercise influence metabolic markers and mRNA expression of ERS-related genes. Weanling C57Bl/6 mice were randomly assigned to receive ad libitum access to standard laboratory chow, AIN93 diet, and our novel “Americanized” diet (AD) for 4-weeks. After 2-weeks, mice from each diet group were randomly assigned to exercise (n=7) or sedentary control groups (n=3). The exercise condition consisted of access to an exercise wheel, three times per week, for 16 hours per day. Wheel utilization was recorded using a bicycle computer/sensor. Insulin sensitivity was determined using standardized insulin-tolerance test. Mice were fasted, euthanized, and blood and liver tissues were collected. Circulating glucose and lipid levels were quantified and hepatic mRNA expression of genes related to ERS ( Ddit3 , Hspa5 , and Erdj4 ) were determined. Data were analyzed using GLM procedures in SPSS. Despite having no influence on body weight, diet (P=0.04) significantly influenced liver weight, with mice fed AD having greater (P = 0.01) liver weight compared to mice fed chow. Exercise tended to increase liver weight compared to the control (P=0.095), while reducing liver lipid content in mice fed chow and AD. AD fed mice had the highest circulating glucose values compared to mice fed chow (P=0.013) and also tended to be greater than mice fed AIN (P=0.056). Exercise significantly increased circulating triglycerides (P=0.03) and tended to reduce circulating glucose levels (P=0.07). Diet significantly influenced insulin sensitivity (P = 0.003), with mice fed AD having less insulin sensitivity as compared to mice fed chow (P&lt;0.001) and also tended to be lower than mice fed AIN (P=0.06). While non-significant (P=0.15), exercising mice tended to have less insulin sensitivity than controls. Exercise caused an increase in Erdj4 mRNA expression (P=0.02), most notably in mice fed AD, which had a greater ER-stress response with exercise than other diet groups (P=0.04). Our data suggests that the physiological responses to acute exercise is dependent upon the quality of diet consumed, and these effects are associated with changes of markers of ERS. Additional studies are needed to further elucidate the mechanisms by which diet, exercise, and ERS interact to influence metabolic health.

Early Low-Grade Inflammation Induced by High-Salt Diet in Sprague Dawley Rats Involves Th17/Treg Axis Dysregulation, Vascular Wall Remodeling, and a Shift in the Fatty Acid Profile.

Unrestricted increased table salt (NaCl) intake is associated with oxidative stress and inflammation, leading to endothelial dysfunction and atherosclerosis. However, data on salt-induced immunomodulatory effects in the earliest phase of salt loading are scarce. In the present study, an animal model of short-term salt loading was employed, including male Sprague Dawley rats consuming a high-salt diet (HSD; 4% NaCl) or standard laboratory chow (low-salt; LSD; 0.4% NaCl) during a 7-day period. The contribution of angiotensin II (ANGII) suppression was tested by adding a group of rats on a high-salt diet receiving ANGII infusions. Samples of peripheral blood/mesenteric lymph node leukocytes, brain blood vessels, and serum samples were processed for flow cytometry, quantitative real-time PCR, total proteome analysis, and multiplex immunoassay. Data analysis revealed the up-regulation of Il 6 gene in the microcirculation of high-salt-fed rats, accompanied by an increased serum level of TNF-alpha cytokine. The high-salt diet resulted in increased proportion of serum mono-unsaturated fatty acids and saturated fatty acids, reduced levels of linoleic (C18:2 ω-6) and α-linolenic (C18:3 ω-3) acid, and increased levels of palmitoleic acid (C16:1 ω-7). The high-salt diet had distinct, lymphoid compartment-specific effects on leukocyte subpopulations, which could be attributed to the increased expression of salt-sensitive SGK-1 kinase. Complete proteome analysis revealed high-salt-diet-induced vascular tissue remodeling and perturbations in energy metabolism. Interestingly, many of the observed effects were reversed by ANGII supplementation. Low-grade systemic inflammation induced by a HSD could be related to suppressed ANGII levels. The effects of HSD involved changes in Th17 and Treg cell distribution, vascular wall remodeling, and a shift in lipid and arachidonic acid metabolism.

Western diet consumption does not impact the rewarding and aversive effects of morphine in male Sprague-Dawley rats

The impacts of high-fat and/or high-sugar diets on opioid-induced effects are well documented; however, little is known about the effect of such diet on the affective responses to opiates. To address this issue, in the present experiment male Sprague-Dawley rats were given ad libitum access to a western-style diet (high in saturated fat and sugar) or a standard laboratory chow diet beginning in adolescence and continuing into adulthood at which point they were trained in a combined conditioned taste avoidance (CTA)/conditioned place preference (CPP) procedure to assess the aversive and rewarding effects of morphine, respectively. On four conditioning cycles, animals were given access to a novel saccharin solution, injected with morphine (1 mg/kg or 5 mg/kg), and then placed on one side of a place preference chamber. Animals were then tested for place preference and saccharin preference. All subjects injected with morphine displayed significant avoidance of the morphine-associated solution (CTA) and preferred the side associated with the drug (CPP). Furthermore, there were no differences between the two diet groups, indicating that chronic exposure to the western diet had no impact on the affective properties of morphine (despite increasing caloric intake, body weight, body fat and lean body mass). Given previously reported increases in drug self-administration in animals with a history of western-diet consumption, this study suggests that western-diet exposure may increase drug intake via mechanisms other than changes in the rewarding or aversive effects of the drug.

Insulin sensitivity is preserved in mice made obese by feeding a high starch diet.

Obesity is generally associated with insulin resistance in liver and muscle and increased risk of developing type 2 diabetes, however there is a population of obese people that remain insulin sensitive. Similarly, recent work suggests that mice fed high carbohydrate diets can become obese without apparent glucose intolerance. To investigate this phenomenon further, we fed mice either a high fat (Hi-F) or high starch (Hi-ST) diet and measured adiposity, glucose tolerance, insulin sensitivity, and tissue lipids compared to control mice fed a standard laboratory chow. Both Hi-ST and Hi-F mice accumulated a similar amount of fat and tissue triglyceride compared to chow-fed mice. However, while Hi-F diet mice developed glucose intolerance as well as liver and muscle insulin resistance (assessed via euglycaemic/hyperinsulinaemic clamp), obese Hi-ST mice maintained glucose tolerance and insulin action similar to lean, chow-fed controls. This preservation of insulin action despite obesity in Hi-ST mice was associated with differences in de novo lipogenesis and levels of C22:0 ceramide in liver and C18:0 ceramide in muscle. This indicates that dietary manipulation can influence insulin action independently of the level of adiposity and that the presence of specific ceramide species correlates with these differences.

Gestational protein restriction alters early amygdala neurochemistry in male offspring

ABSTRACT Background Gestational protein intake restriction-induced long-lasting harmful outcomes in the offspring's organs and systems. However, few studies have focused on this event's impact on the brain's structures and neurochemical compounds. Aim The present study investigated the effects on the amygdala neurochemical composition and neuronal structure in gestational protein-restricted male rats' offspring. Methods Dams were maintained on isocaloric standard rodent laboratory chow with regular protein [NP, 17%] or low protein content [LP, 6%]. Total cells were quantified using the Isotropic fractionator method, Neuronal 3D reconstruction, and dendritic tree analysis using the Golgi–Cox technique. Western blot and high-performance liquid chromatography performed neurochemical studies. Results The gestational low-protein feeding offspring showed a significant decrease in birth weight up to day 14, associated with unaltered brain weight in youth or adult progenies. The amygdala cell numbers were unchanged, and the dendrites length and dendritic ramifications 3D analysis in LP compared to age-matched NP progeny. However, the current study shows reduced amygdala content of norepinephrine, epinephrine, and dopamine in LP progeny. These offspring observed a significant reduction in the amygdala glucocorticoid (GR) and mineralocorticoid (MR) receptor protein levels. Also corticotrophin-releasing factor (CRF) amygdala protein content was reduced in 7 and 14-day-old LP rats. Conclusion The observed amygdala neurochemical changes may represent adaptation during embryonic development in response to elevated fetal exposure to maternal corticosteroid levels. In this way, gestational malnutrition stress can alter the amygdala's neurochemical content and may contribute to known behavioral changes induced by gestational protein restriction.

Abstract P3067: Sleep Deprivation In Obesogenic Setting Alters Lipidome And Microbiome Toward Suboptimal Inflammation In Acute Heart Failure

Introduction: Sleep is fundamental medicine for cardiac homeostasis and the immune fitness of human health. Sleep-deprived individuals are prone to higher incidences of a heart attack. The lipid-dense diet (obesogenic diet-OBD) is a cumulative risk factor for chronic inflammation in cardiovascular disease, thus understanding how sleep fragmentation (SF) in obesity setting impacts immune and cardiac health is an unmet medical need to develop novel therapeutics. Hypothesis: We hypothesized whether the co-existence of SF with OBD dysregulates gut homeostasis and leukocyte-derived reparative mediators thereby impairing cardiac repair. Methods: Two months old male C57BL/6J mice were randomized into two main groups; control (fed standard lab chow) and OBD (fed 10% w/w safflower oil-enriched diet) for 2 months. For SF, mice were subjected to SF chamber (2-minute interval bar sweep in light period (0700–1900 h) for 3 days. Mice of control, control+SF, OBD control, OBD+SF groups were further subjected to permanent coronary artery ligation to induce acute heart failure. Results: OBD fed mice had higher levels of plasma linolenic acid with a decrease in eicosapentaenoic acid and docosahexaenoic acid. The OBD mice had lower levels of Lactobacillus johnsonii indicating loss of probiotic microbiota. SF in OBD mice increased Firmicutes/Bacteroidetes (F/B) ratio indicative of a detrimental change in SF-directed microbiome signatures. OBD+SF group increased in the neutrophil: lymphocyte ratio suggestive of suboptimal inflammation. As a result of SF, resolution mediators (RvD2, RvD3, RvD5, LXA 4 , PD1, and MaR1) decreased and inflammatory mediators (PGD 2 , PGE 2 , PGF 2a , 6k-PGF 1a ) were increased post- MI in OBD fed mice. At the site of infarction, the proinflammatory cytokines; Ccl2, IL1β , and IL-6 were amplified in OBD+SF indicating a robust proinflammatory milieu. OBD+SF group showed signs of neutropenia in the spleen with pre-activation of CD169 macrophages. Also, brain circadian genes ( Bmal1, Clock) were downregulated in SF subjected control mice but remained elevated in OBD mice post-MI. Conclusion: SF superimposed on obesity dysregulated physiological inflammation and disrupted resolving response thereby impaired cardiac repair and signs of pathological inflammation.

Chrysin reduces hypercholesterolemia-mediated atherosclerosis through modulating oxidative stress, microflora, and apoptosis in experimental rats.

Chrysin (Chy) is known for various biological proprieties such as inhibitory effects on inflammation, cancer, oxidative stress, aging, and atherosclerosis. However, the hypolipidemic activity of Chy and its mechanistic action remains unclear in cardiovascular diseases (CVD). In this study, we focused on the hypolipidemic proprieties of Chy in hypercholesterolemia-induced atherosclerosis. Male Wistar rats (150-220 g) were divided into four groups as follows: Group I control was fed with standard laboratory chow. Rats in Group II were fed a high-fat diet (HFD) for 60 days. After 60 days of HFD, Group III rats received Chy (100 mg/kg body weight); Group IV rats received Atorvastatin (Atv; 10mg/kg body weight) for 30 days. Biochemical studies showed Chy, Atv treatment decreased the activities of liver marker enzymes and the levels of Reactive Oxygen Species (ROS) and lipid profile. Gene expression analysis on nuclear factor erythroid 2-related factor 2 (Nrf2) and its regulated genes were significantly reduced in the intestine and increased in the aorta by Chy and Atv. Gut microbial species such as Bacteroidetes, Lactobacillus, Enterococcus, and Clostridium leptum copy numbers were significantly increased by Chy and Atv treatment. In addition, Chy and Atv modulated the expression of inflammatory genes including TLR4, TNFα, NLRP3, and IL-17 in the aorta and intestine compared with hypercholesterolemic control rats. Chy and Atv effectively increased the caspase-3 mRNA expression in the intestine, but these decreased in the aorta. The present study concludes that by reducing oxidative stress and increasing gut microbial colonization, Chy may provide an effective therapeutic approach for the prevention of hypercholesterolemia-mediated atherosclerosis. PRACTICAL APPLICATIONS: Our study focused on a therapeutic model representing the clinical presentation of atherosclerosis in humans. Statins are commonly used in the treatment of cardiovascular complications, patients with hypercholesterolemia face difficulties in the continuation of statin therapy. The reason for statin discontinuation has been associated with toxicological effects. It is necessary to investigate the potentiality of the natural compound as an alternative medicine to statin with fewer side effects. The main theme of our study is to compare the therapeutic potential of Chy and Atv. Chy is a natural bioflavonoid that could be considered as an alternative medicinal compound to statins and to avoid toxicity problems associated with statins. Chy is a bioflavonoid present in Passiflora caerulea (blue passion flower), Oroxylum indicum (Indian trumpet flower), Pelargonium crispum, propolis, and honey. Consuming Chy-rich foods will reduce hypercholesterolemia-mediated cardiovascular complications. Overall, the present studies provided a key to developing bioactive compounds-based foods for CVD patients.

Glutamine energy substrate anaplerosis increases bone density in the Pahenu2 classical PKU mouse in the absence of phenylalanine restriction.

Osteopenia is an under‐investigated clinical presentation of phenylalanine hydroxylase (PAH)‐deficient phenylketonuria (PKU). While osteopenia is not fully penetrant in human PKU, the Pahenu2 mouse is universally osteopenic and ideal to study the phenotype. We determined Pahenu2 mesenchymal stem cells (MSCs) are developmentally impaired in the osteoblast lineage. Moreover, we determined energy dysregulation and oxidative stress contribute to the osteoblast developmental deficit. The MSC preferred substrate glutamine (Gln) was applied to enhance energy homeostasis. In vitro Pahenu2 MSCs, in the context of 1200 μM Phe, respond to Gln with increased in situ alkaline phosphatase activity indicating augmented osteoblast differentiation. Oximetry applied to Pahenu2 MSCs in osteoblast differentiation show Gln energy substrate increases oxygen consumption, specifically maximum respiration and respiratory reserve. For 60 days post‐weaning, Pahenu2 animals received either no intervention (standard lab chow), amino acid defined chow maintaining plasma Phe at ~200 μM, or standard lab chow where ad libitum water was a 2% Gln solution. Bone density was assessed by microcomputed tomography and bone growth assessed by dye labeling. Bone density and dye labeling in Phe‐restricted Pahenu2 was indistinguishable from untreated Pahenu2. Gln energy substrate provided to Pahenu2, in the context of uncontrolled hyperphenylalaninemia, present increased bone density and dye labeling. These data provide further evidence that Pahenu2 MSCs experience a secondary energy deficit that is responsive both in vitro and in vivo to Gln energy substrate and independent of hyperphenylalaninemia. Energy support may have effect to treat human PKU osteopenia and elements of PKU neurologic disease resistant to standard of care systemic Phe reduction. Glutamine energy substrate anaplerosis increased Pahenu2 bone density and improved in vitro MSC function in the context of hyperphenylalaninemia in the classical PKU range.

Influence of Maternal Diet and Intergenerational Change in Diet Type on Ovarian and Adipose Tissue Morphology in Female Rat Offspring.

Background and Objectives: A high-fat diet causes inflammation in the organism and many metabolic disorders. Adipose tissue secretes adipokines that affect the function of many organs. The health status of the mother before and during pregnancy affects the health of the offspring. The aim of this study was to determine how the type of maternal diet and the change in the type of diet in the offspring affects the histological characteristics of the ovaries and subcutaneous and perigonadal adipose tissue in female rat offspring. Materials and Methods: Ten female rats were divided into two groups. One group was fed standard laboratory chow, and the other was fed a high-fat diet and mated with a male of the same breed. The offspring of both groups of dams were divided into four subgroups with different feeding protocols. At 22 weeks of age, the offspring were sacrificed. Ovaries and subcutaneous and perigonadal adipose tissue were isolated. In the ovaries, the presence of cystic formations was investigated. Histomorphometric analysis was performed in two types of adipose tissue. Results: The weight of the ovaries of the offspring of mothers fed a high-fat diet was significantly higher than that of the offspring of mothers fed standard laboratory diets. Cystic formations were found in the ovaries of the offspring of mothers fed a high-fat diet. In subcutaneous adipose tissue, the percentage of small-sized adipocytes was significantly higher in the offspring of mothers fed standard laboratory diets. There were no significant differences in adipocyte surface area and adipocyte number between groups. Conclusion: Maternal diet influences the morphology of the ovaries and adipose tissue of the offspring.

Eating a High Fat Diet Enhances Sensitivity of Rats to 5‐HT1A Receptor‐Mediated Behavioral Effects of Serotonergic Drugs

Diet (e.g., type and amount of food consumed) has been shown to impact drug sensitivity, in ways that might be relevant for the therapeutic effects and side effects of medications. For example, previous studies have demonstrated that rats eating a high fat diet are more sensitive to some of the effects of dopaminergic drugs. Considerably less is known about how eating a high fat diet might impact sensitivity to drugs that act on serotonin (5‐HT) systems, many of which might be beneficial for the treatment of obesity, since they can decrease feeding. Drugs with agonist effects at 5‐HT receptors can produce a collection of adverse effects, known as 5‐HT syndrome. Animal models of 5‐HT syndrome include lower lip retraction, flat body posture, penile erections, and forepaw treading. To test the hypothesis that eating a high fat diet enhances sensitivity of rats to 5‐HT syndrome, male (n=16) and female (n=16) rats eating high fat (60% kcal from fat) or standard (17% kcal from fat) laboratory chow were tested once weekly with cumulative doses of 5‐HT receptor agonists, including 8‐OH‐DPAT (0.01‐1.0 mg/kg, s.c.), lorcaserin (1.0‐32.0 mg/kg, i.p.) and WAY 163909 (1.0‐32.0 mg/kg, i.p.). Agonists were also studied in combination with antagonists selective for 5‐HT1A (WAY 100635; 0.178 mg/kg, s.c.), or 5‐HT2C (SB 242084; 1.0 mg/kg; i.p.) receptors. Food consumption and body weight were measured throughout the study. Results were analyzed using either two‐way or three‐way mixed model ANOVAs with diet and dose, diet and day, or diet, sex, and drug as factors, and Bonferroni post hoc comparisons where appropriate. Effects mediated by the 5‐HT2C receptor (e.g, penile erections as well as decreased food consumption), were only observed following injections of lorcaserin and WAY 163909 (but not 8‐OH‐DPAT), and were not different between rats eating standard or high fat chow. Regarding 5‐HT1A receptor‐mediated effects, all three 5‐HT receptor agonists induced forepaw treading in rats; however, flat body posture and lower lip retraction were only induced by 8‐OH‐DPAT. Rats eating high fat chow were more sensitive to forepaw treading, regardless of which drug induced this behavior; however, WAY 163909 induced less forepaw treading as compared to the other two drugs, regardless of diet. Rats eating high fat chow were also more sensitive to 8‐OH‐DPAT‐induced lower lip retraction, but not flat body posture. These results suggest that eating a high fat diet can impact sensitivity of rats to some of the effects of serotonergic drugs. Additionally, these results demonstrate that while both lorcaserin and WAY 163909 decrease feeding, WAY 163909 produces less 5‐HT1A‐mediated forepaw treading than lorcaserin, which might indicate that it will produce fewer side effects in humans as well. Finally, these results suggest that dietary history could influence sensitivity of individuals to the adverse effects of serotonergic drugs, such as 5‐HT syndrome.

PHB Improves Metabolic Homeostasis in Obese C57BL6 Male Mice Fed a High Fat Diet

Pharmacological approaches to combat metabolic syndrome have been met with limited success. Given the critical role AMPK plays in metabolic homeostasis, targeting this protein as a potential treatment and prevention strategy for metabolic syndrome could be of clinical interest. Our current study investigated the effects of a well‐defined poly‐herbal blend (PHB) on high fat diet (HFD)‐induced body mass dysregulation and alterations in metabolic homeostasis and glycemic control in mice. Age‐matched mice were fed standard lab chow for six weeks, then switched to HFD supplemented with PHB. Preliminary data demonstrate that body weights of control and PHB‐treated mice were comparable in chow fed animals. However, mice fed HFD supplemented with PHB gained less weight compared to controls. Furthermore, PHB‐fed obese mice exhibited improved glucose tolerance and insulin sensitivity. Food intake of control and PHB‐treated mice were comparable on both diets. Consistent with the decreased body weight, PHB supplementation resulted in decreased fat mass. Mechanistic studies in adipocytes and muscle cells demonstrated that these beneficial effects were mediated, at least in part, through regulation of the AMPK signaling pathway. Taken together, our preliminary studies identify PHB as a natural and sustainable approach for the treatment and prevention of metabolic syndrome and warrant additional investigation.