Diet-resistant Mice Research Articles

High-fat diet and dyslipidemia synergistically contribute to T cell senescence in gut associated lymphoid tissue.

Obesity can compromise immune response and immune surveillance. Recent studies have linked obesity with systemic T cell senescence and thymus involution. However, these studies failed to distinguish the influence of obesity from the influence of diet composition on premature T cell senescence. High-fat diet (HFD) can influence the immunity of gut-associated lymphoid tissue (GALT) preceding the onset of obesity. Despite GALT is sensitive to the changes in the dietary composition and metabolic status, it still remains elusive how HFD and obesity contribute to T cell senescence in the GALT. In this study, we illustrated the interplay of the HFD, obesity and intestinal immunity by comparing the immune features of diet-induced obese (DIO) to those of diet-resistant (DR) mice. As expected, DIO mice exhibited increased serum lipid levels and liver steatosis whereas dyslipidemia was absent in DR mice. DIO mice demonstrated a shift from naïve to effector-memory (EM) phenotype in the T cells derived from the spleens and PPs. Moreover, DIO mice showed up-regulation of PD1 and KLRG1 on the T cells. Similar but mild trends were observed in the naïve and EM T cells from DR mice. Furthermore, we proved that the senescent-like changes in splenic and PPs-derived T cells positively correlated with the serum lipid concentrations. Taken together, our results indicate that HFD components function synergistically with dyslipidemia to induce T cell senescence in the GALT.

The Effect of Diet-Induced and Melanocortin Obesity on Expression of Tryptophan Hydroxylase 2 in the Dorsal Raphe Nucleus and Ventral Tegmental Area in Mice

Serotonin and its signaling pathways are involved in the central regulation of feeding behavior and energy metabolism. The most important role in this regulation is played by dopaminergic neurons of the dorsal raphe nucleus (DRN) and ventral tegmental area (VTA) which express tryptophan hydroxylase 2 (TPH2) catalyzing the rate-limiting step in serotonin synthesis. Changes in activity of the serotonin-synthesizing system in the DRN and VTA may significantly contribute to the development of metabolic disorders, including obesity. However, data on TPH2 expression in these brain regions in obesity are unavailable. The aim of this work was to compare immunohistochemically TPH2 expression in the DRN and VTA of C57Bl/6J (a/a) female mice with a diet-induced obesity (DIO) (caused by 8- and 16-week intake of high-calorie food) and C57Bl/6J (Ay/a) agouti mice with genetically predetermined melanocortin obesity. Double immunolabeling demonstrated that in DIO mice with a 16-week high-calorie diet TPH2 expression in the DRN (the main cerebral source of serotonin) decreases while in the VTA increases. In agouti mice, TPH2 expression in the DRN did not change while in the VTA increased. In DIO mice with a 8-week high-calorie diet and in C57Bl/6J (a/a) mice resistant to this diet, TPH2 expression did not change both in the DRN and VTA. Thus, DIO mice kept on a long-term (16 weeks) diet and agouti mice with melanocortin obesity exhibit specific changes in the serotonin synthesis that are lacking in diet-resistant mice. This finding indicates a crucial role of serotonin signaling in the organismal response to excessive caloric intake.

Alterations to the microbiota–colon–brain axis in high-fat-diet-induced obese mice compared to diet-resistant mice

Obesity is underpinned by both genetic and environmental factors, including a high-saturated-fat diet. Some mice develop diet-induced obesity (DIO), but others remain diet resistant (DR) despite intake of the same high-saturated-fat diet, a phenomenon that mimics characteristics of the human obese phenotype. Microbiota–colon–brain axis regulation is important for energy metabolism and cognition. Using DIO and DR mouse models, this study aimed to examine gut microbiota, colonic inflammation and cognitive function to elucidate the role of microbiota–gut–brain regulation in DIO. C57Bl6/J mice fed a chronic saturated-palmitic-acid diet for 22 weeks showed significant body weight gain differences, with the top one third gaining 48% heavier body weight than the lower one third. There was significant reduction in gut microbiota richness and diversity in DIO mice but not in DR mice. At the phylum level, DIO mice had increased abundance of Firmicutes and Antinobacteria, and decreased abundance of Bacterioides and Proteobacteria in gut microbiota. DIO mice exhibited reduced tight junction proteins, increased plasma endotoxin lipopolysaccharide (LPS) and increased inflammation in the colon and liver. Recognition memory and spatial memory were impaired in DIO mice, associated with decreased Bacteroidetes. Further examination showed that hippocampal brain-derived neurotrophic factor was significantly decreased in DIO mice (vs. DR). Conversely, DR mice showed no changes in the above parameters measured. Therefore, gut microbiota, colon inflammation and circulating LPS may play a major role in the development of the obese phenotype and cognitive decline associated with a chronic high-saturated-palmitic-acid diet.

Resistin Is Associated with Blood-Brain Barrier Disruption in Mice Resistant to Diet-Induced Obesity and Treated with Topiramate

Resistin has been linked to diabetes, obesity, insulin resistance, atherosclerosis, and the development of cardiovascular disease. However, the effects and the molecular mechanisms of resistin on endothelial permeability, a key event in the development of atherosclerosis, inflammation, and vascular disease, have not been fully elucidated. Diabetes is associated with changes in the brain microvasculature leading to dysfunction and disruption of the blood-brain barrier (BBB), a specialized system of microvascular endothelial cells protecting the brain from toxic substances in circulation, while permitting CNS entry of nutrients and endocrine signals using active transport mechanisms and passive diffusion. As diabetes is associated with cognitive impairment, understanding the molecular mechanisms of endothelial permeability in the brain is important in developing appropriate therapeutic agents. Topiramate (TPM), a mitochondrial carbonic anhydrase inhibitor, prevents BBB disruption in mouse models of type I and II diabetes. Mice resistant to the development of diet-induced obesity, when treated with TPM, had increased serum levels of resistin and increased BBB permeability to all brain regions except the hippocampus and hypothalamus, brain regions paradoxically disrupted in models of type II diabetes. In vitro treatment of brain endothelial cells with resistin demonstrated a decrease in endothelial nitric oxide synthase (eNOS) along with increased permeability and decreased TEER, a measure of electrical resistance across a cell monolayer. This increase in permeability occurs because diet-resistant mice treated with TPM are unable to attenuate the effects of hyperglycemia-induced oxidative stress and inflammation. Topiramate, which is protective of the BBB in types I and II diabetes, seems paradoxically toxic to the BBB in mice resistant to diet-induced obesity and this toxicity is associated with increases in serum resistin. Disclosure T.S. Salameh: None. W. Mortell: None. W.A. Banks: None.

Depot-specific differences in angiogenic capacity of adipose tissue in differential susceptibility to diet-induced obesity

ObjectiveAdipose tissue (AT) expansion requires AT remodeling, which depends on AT angiogenesis. Modulation of AT angiogenesis could have therapeutic promise for the treatment of obesity. However, it is unclear how the capacity of angiogenesis in each adipose depot is affected by over-nutrition. Therefore, we investigated the angiogenic capacity (AC) of subcutaneous and visceral fats in lean and obese mice. MethodsWe compared the AC of epididymal fat (EF) and inguinal fat (IF) using an angiogenesis assay in diet-induced obese (DIO) mice and diet-resistant (DR) mice fed a high-fat diet (HFD). Furthermore, we compared the expression levels of genes related to angiogenesis, macrophage recruitment, and inflammation using RT-qPCR in the EF and IF of lean mice fed a low-fat diet (LFD), DIO mice, and DR mice fed a HFD. ResultsDIO mice showed a significant increase in the AC of EF only at 22 weeks of age compared to DR mice. The expression levels of genes related to angiogenesis, macrophage recruitment, and inflammation were significantly higher in the EF of DIO mice than in those of LFD mice and DR mice, while expression levels of genes related to macrophages and their recruitment were higher in the IF of DIO mice than in those of LFD and DR mice. Expression of genes related to angiogenesis (including Hif1a, Vegfa, Fgf1, Kdr, and Pecam1), macrophage recruitment, and inflammation (including Emr1, Ccr2, Itgax, Ccl2, Tnf, and Il1b) correlated more strongly with body weight in the EF of HFD-fed obese mice compared to that of IF. ConclusionsThese results suggest depot-specific differences in AT angiogenesis and a potential role in the susceptibility to diet-induced obesity.

High fat diet-induced obesity increases the formation of colon polyps induced by azoxymethane in mice.

Obesity has been found to be associated with colon cancer. However, the mechanism of this relationship is unclear and thus a good animal model is required. Our previous research showed that some mice developed diet-induced obesity (DIO) whilst others were diet-resistant (DR) when fed a high-fat diet. In the present study, we have tested the effects of a high-fat diet on the formation of colon polyps induced by azoxymethane (AOM) in both DIO and DR mice. We found that the DIO mice have developed 2.5 times of polyps compared to the DR mice (P<0.05) and 3.4 times of polyps compared to the low fat fed mice (P<0.05). Although the DR mice tended to have more polyps than the low-fat diet fed mice, this was not statistically significant. The DIO mice could have an increased polyp formation due to obesity-related cancer risk factors and different gene expression from DR mice. DIO mice could be used as an appropriate model for studying obesity-associated colon cancer; however DR mice are not suitable because they don't show any significant weight gains to indicate obesity.

Increased oxidative stress and the apoptosis of regulatory T cells in obese mice but not resistant mice in response to a high-fat diet

High fat feeding induces a variety of obese and lean phenotypes in inbred rodents. Obesity is a pro-inflammatory state, and regulatory T cells (Tregs) are essential negative regulators of inflammation. We aimed to determine the involvement of Tregs in the mice susceptible or resistant to high-fat diet (HFD). In the study, diet-induced obese (DIO) mice experienced significant increases in weight gain, energy intake, fat masses, plasma lipid and proinflammatory cytokines in comparison with control and diet-resistant (DR) mice. Also, Tregs production decreased in DIO mice. HFD diminished mitochondrial transmembrane potential (MTP) in the spleen Tregs of DIO mice and reinforced apoptosis compared with that in DR mice. Moreover, HFD significantly decreased antioxidant enzymes expressions and increased reactive oxygen species (ROS) productions in the Tregs of DIO mice, but not in those of DR mice, which should provide valuable evidence for unraveling the pathogenesis of inflammation found in this obese mice model.

Resveratrol prevents suppression of regulatory T-cell production, oxidative stress, and inflammation of mice prone or resistant to high-fat diet–induced obesity

Consumption of a high-fat diet (HFD) is correlated with increased oxidative stress and chronic inflammation in many organs. Regulatory T cells (Tregs) are essential negative regulators of inflammation. We hypothesized that resveratrol (trans-3,5,4′-trihydroxystilbene) could protect against HFD-induced oxidative stress and inflammation. Therefore, we examined the effect of resveratrol on oxidative stress and the relevant peripheral immune-regulating mechanisms in HFD-induced obese (DIO) and diet-resistant mice. C57BL/6 mice were fed a normal diet and an HFD for 13 weeks. Then the experimental group was subdivided into DIO and diet-resistant groups according to their body weights, which were further supplemented with 0.03% resveratrol and 0.06% resveratrol, respectively, for an additional 13 weeks. Resveratrol prevented the accumulation of chronic oxidative stress and suppression of Tregs production in HFD mice, modulated changes of cytokines in the plasma and spleen, and decreased expressions of inflammatory mediators compared with those of the DIO group. Our results indicate that resveratrol, as a feasible effective supplement for HFD, can relieve oxidative stress, inhibit inflammatory genes expression, and increase Tregs number via aryl hydrocarbon receptor activation inhibited by HFD, especially in DIO mice.

Pharmacological Modulation of Dopamine Receptor D2-Mediated Transmission Alters the Metabolic Phenotype of Diet Induced Obese and Diet Resistant C57Bl6 Mice

High fat feeding induces a variety of obese and lean phenotypes in inbred rodents. Compared to Diet Resistant (DR) rodents, Diet Induced Obese (DIO) rodents are insulin resistant and have a reduced dopamine receptor D2 (DRD2) mediated tone. We hypothesized that this differing dopaminergic tone contributes to the distinct metabolic profiles of these animals. C57Bl6 mice were classified as DIO or DR based on their weight gain during 10 weeks of high fat feeding. Subsequently DIO mice were treated with the DRD2 agonist bromocriptine and DR mice with the DRD2 antagonist haloperidol for 2 weeks. Compared to DR mice, the bodyweight of DIO mice was higher and their insulin sensitivity decreased. Haloperidol treatment reduced the voluntary activity and energy expenditure of DR mice and induced insulin resistance in these mice. Conversely, bromocriptine treatment tended to reduce bodyweight and voluntary activity, and reinforce insulin action in DIO mice. These results show that DRD2 activation partly redirects high fat diet induced metabolic anomalies in obesity-prone mice. Conversely, blocking DRD2 induces an adverse metabolic profile in mice that are inherently resistant to the deleterious effects of high fat food. This suggests that dopaminergic neurotransmission is involved in the control of metabolic phenotype.

Differential expression of hypothalamic CART mRNA in response to body weight change following different dietary interventions

Cocaine- and amphetamine-regulated transcript (CART) peptide is widely expressed in the hypothalamus and is involved in the central regulation of energy balance. Using in situ hybridization, this study examined the roles of CART peptide in the hypothalamus of diet-induced obese (DIO) or diet-resistant (DR) mice under different dietary interventions including high-fat (HF), low-fat (LF) and pair-feeding (PF) diet for 6 weeks. Pair feeding the energy intake of the DIO and DR mice was used to determine whether there is an inherent difference in baseline CART expression that may cause the DIO and DR phenotypes. The results demonstrated that CART mRNA expression in the hypothalamus of the DIO mice responded differently on the high-fat diet compared to DR mice. The arcuate nucleus and paraventricular nucleus showed a significant reduction in CART mRNA expression in DIO mice compared to DR mice on the HF diet (−19.6%, p = 0.019; −26.1%, p = 0.003); whilst a profound increase in CART mRNA expression was observed in the dorsomedial nucleus and lateral hypothalamic area (+44.5%, p = 0.007; +37.4%, p = 0.033). Our study suggests that the decrease of CART mRNA expression in Arc and PVN regions of DIO mice may contribute to the development of high-fat diet-induced obesity. In addition, CART in the dorsomedial nucleus (DM) of hypothalamus and lateral hypothalamus (LH) may be involved in the activation of an orexigenic effect. Since pair feeding of the high-fat diet eliminated both the body weight and CART mRNA differences between the DIO and DR mice, it is likely that their alterations in gene expression were a consequence of their dissimilar body weight levels.

Ventromedial Hypothalamic NPY Y2 Receptor in the Maintenance of Body Weight in Diet-Induced Obesity in Mice

This study examined changes in neuropeptide Y (NPY) Y2 receptor binding in the brains of C57BL/6 mice in response to different levels of high-fat diets via three dietary intervention methods: high-fat diet, switching from high- to low-fat diet and finally, energy restricted high-fat diet. Forty-five C57Bl/6 male mice were fed a high-fat diet for 8 weeks and then classified as diet-induced obese (DIO) or diet-resistant (DR) mice according to the highest and lowest body weight gainers, respectively. The DIO and DR mice were then randomly divided into three groups each and either continued on their high-fat diet ad libitum (DIO-H and DR-H), changed to a low-fat diet (DIO-L and DR-L) or pair-fed via energy restricted high-fat diet (DIO-P and DR-P) for a further 6 weeks. During the course of this study, body weight, energy intake and plasma peptide YY (PYY) were measured. The study revealed that the replacement of a high-fat diet with a low-fat diet was associated with a significant lowering of ventromedial hypothalamic (VMH) Y2 receptor binding in both the DIO-L and DR-L mice (-37%, -36%), and also a lowered plasma PYY level in the DIO-L mice (-25%). Despite a continued consumption of the high-fat diet, energy restricted pair feeding caused a lower VMH Y2 receptor binding in the obese mice (DIO-P) following weight loss compared to the DR-P mice (-14%). In conclusion, this study showed that changing diets from high- to low-fat can significantly lower the VMH Y2 receptor binding irrespective to the obesity phenotype. Energy restriction, even while on high-fat feeding, can cause a lower VMH Y2 receptor binding compared to DR mice even after body weight loss to similar levels. This suggests either a possible intrinsic nature of the DIO mice or a body weight set-point re-establishment to drive body weight regain.

Role of fat amount and type in ameliorating diet-induced obesity: insights at the level of hypothalamic arcuate nucleus leptin receptor, neuropeptide Y and pro-opiomelanocortin mRNA expression.

Dietary fatty acid profile, independent of caloric percent of fat, is a major regulator of body adiposity. This study examined the effects of dietary fat amount and types on fat storage and hypothalamic gene expression in the mouse model of chronic diet-induced obesity. The dietary interventions were in twofold: (1) the obesity was induced by a 13-week obesogenic fat diet compared with a low-fat (LF) diet, and (2) the reversibility was tested by using high n-3 polyunsaturated fat (PUFA) and LF diets. Fifty-four C57Bl/6 mice were fed a high-fat (59% in kcal) diet for 13 weeks and then classified as diet-induced obese (DIO) or diet-resistant (DR) mice according to upper and lower tertiles of body weight gain. The DIO mice were then subdivided into three groups for a 6-week secondary dietary intervention. Two of the groups were switched to either a high n-3 PUFA (DIO-n3) or a low-fat (10% in kcal, DIO-LF) diet, whereas the third (controls) and DR mice continued on the initial high-fat diet. Food efficiency was calculated as weekly body weight gain per gram of food intake. After switching the DIO mice to the n-3 PUFA or LF diet, their body weights were reduced to the level of the DR and LF mice. The food efficiencies were, from the highest to lowest, in the order: DIO>LF>DR>DIO-LF>DIO-n3. Using quantitative in situ hybridization, we found that the DIO mice had higher levels of leptin receptor (LR, +290%, p<0.005) and neuropeptide Y (NPY, +25%, p<0.05) mRNA expression in the hypothalamic arcuate nucleus (Arc) than the DR mice, whereas the level of pro-opiomelanocortin (POMC) mRNA expression was significantly reduced (-45%, p<0.01). All effects that were essentially returned to DR levels by the change to the n-3 PUFA diet and, with the exception of a failure to normalize Arc NPY mRNA levels, by the change to LF diet. Taken together, the present results show that both change in level and quality of dietary fat can potently alter hypothalamic neuropeptide expression and result in effective amelioration of diet-induced obesity. Interestingly, the n-3 PUFA diet when fed to already obese mice produced a pattern of hypothalamic gene expression similar to that in obesity resistant (DR) mice. It remains to be determined if the effects of n-3 fatty acids on brain neuropeptide gene expression are direct or indirect.

Altered levels of POMC, AgRP and MC4-R mRNA expression in the hypothalamus and other parts of the limbic system of mice prone or resistant to chronic high-energy diet-induced obesity

The melanocortinergic system plays an important role in promoting negative energy balance and preventing excessive fat deposition. This study has investigated the levels of mRNA expression of proopiomelanocortin (POMC), agouti-related protein (AgRP) and the melanocortin-4 receptor (MC4-R) in diet-induced obese (DIO) and diet-resistant (DR) mice. Thirty C57 mice were used in this study. Twenty-four mice were fed with a high-fat diet (HF: 40% of calories from fat, 20% from saturated fat) for 4 weeks and then classified as DIO and DR according to their body weight gain. Six mice were placed on a low-fat diet (LF: 10% of calories from fat, 1% from saturated fat) and were used as controls. After 22 weeks of feeding, visceral fat deposits were more than twice as heavy in the DIO mice as in the DR and LF mice, while the latter two groups had no significant difference. Using quantitative in situ hybridization techniques, this study found that the DIO mice had a significantly lower level of Arc POMC (−29%) and AgRP (−31%) mRNA expression than the DR and LF mice, respectively. The mice on high-fat diets had higher levels of AgRP mRNA expression in the bed nucleus of stria terminalis (BST), and ventral part of the lateral septal nucleus (LSV) than the LF mice. Furthermore, the DIO mice had a 40% higher level of MC4-R mRNA expression in the ventromedial hypothalamic nucleus (VMH) and posterodorsal part of the medial amygdaloid nucleus (MePD) than the LF mice. In conclusion, this study has demonstrated that differential expression of POMC, AgRP and MC4-R mRNA levels exists in DIO, DR and LF mice. These differences were shown to occur in the specific nuclei of the hypothalamus and other parts of the limbic system. These findings may assist in understanding the involvement of the melanocortinergic system in the regulation of body weight via the autonomic and limbic systems.

Hypothalamic c- fos-like immunoreactivity in high-fat diet-induced obese and resistant mice

Some C57Bl/6 mice become obese, whereas others remain lean when raised on a high-fat diet. The mechanisms underlying this interindividual susceptibility to diet-induced obesity remain unknown. Because hypothalamus plays a major role in the regulation of body weight, this study was conducted to identify the differences of hypothalamic neuronal activity between diet-induced obese and diet-resistant mice. Using c- fos as a marker, this study showed that diet-induced obese mice significantly increased c- fos-like immunoreactive neurons in the dorsal part of lateral hypothalamus (+183%) and dorsomedial hypothalamic nucleus (+87.5%) compared with diet-resistant mice. Furthermore, switching from high fat to low fat, or high n-3 polyunsaturated fatty acid diet, significantly decreased body weight gain (−35.7% and −31.0%), overall fat storage (−63.4% and −59.6%), and c- fos-like immunoreactive neurons in the dorsal part of lateral hypothalamus (−76.5% and −64.7%) and dorsomedial hypothalamic nucleus (−73.3% and −56.7%) in diet-induced obese mice, respectively. The present study also showed that the ratio of serum leptin/fat mass was threefold higher in the diet-resistant mice than in the diet-induced obese mice, which may be responsible for the less fat storage in the diet-resistant mice. The current data further confirm that the increased neuronal activity in the key autonomic regulatory centers may contribute to the excessive fat storage in diet-induced obese mice. Moreover, both high-fat diet-induced excessive fat storage and the altered hypothalamic neuronal activity may be largely corrected by reducing dietary fat content or replacing it with non-obesogenic fat.