Obese Minipigs Research Articles

Respiratory internal kinematics of the tongue base and soft palate in obese minipigs with obstructive sleep apnea.

It is largely unknown how the tongue base and soft palate deform to alter the configuration of the oropharyngeal airway during respiration. This study is to address this important gap. After live sleep monitoring of five Yucatan and two Panepinto minipigs to verify obstructive sleep apnea (OSA), eight and four ultrasonic crystals were implanted into the tongue base and soft palate to circumscribe a cubic and square region, respectively. The 3D and 2D deformational changes of the circumscribed regions were measured simultaneously with electromyographic activity of the oropharyngeal muscles during spontaneous respiration under sedated sleep. The results indicated that both obese Yucatan and Panepinto minipigs presented spontaneous OSA, but not in threenonobese Yucatan minipigs. During inspiration, the tongue base showed elongation in both dorsal and ventral regions but thinning and thickening in the anterior and posterior regions, respectively. The widths showed opposite directions, widening in the dorsal but narrowing in the ventral regions. The soft palate expanded in both length and width. Compared to normal controls, obese/OSA ones showed similar directions of deformational changes, but the magnitude of change was two times larger in the tongue base and soft palate, and obese/OSA Panepinto minipigs presented 10 times larger changes in all dimensions of both the tongue base and the soft palate. The distance changes between the dorsal surface of tongue base and soft palate during inspiration increased in normal but decreased in obese OSA minipigs.

Momordica charantia fruit reduces plasma fructosamine whereas stems and leaves increase plasma insulin in adult mildly diabetic obese Göttingen Minipigs.

Traditionally Momordica charantia (Bitter gourd) is known for its blood glucose lowering potential. This has been validated by many previous studies based on rodent models but human trials are less convincing and the physiological mechanisms underlying the bioactivity of Bitter gourd are still unclear. The present study compared the effects of whole fruit or stems-leaves from five different Bitter gourd cultivars on metabolic control in adult diabetic obese Göttingen Minipigs. Twenty streptozotocin-induced diabetic (D) obese Minipigs (body weight ~85 kg) were subdivided in mildly and overtly D pigs and fed 500 g of obesogenic diet per day for a period of three weeks, supplemented with 20 g dried powdered Bitter gourd or 20 g dried powdered grass as isoenergetic control in a cross-over, within-subject design. Bitter gourd fruit from the cultivars "Palee" and "Good healthy" reduced plasma fructosamine concentrations in all pigs combined (from 450±48 to 423±53 and 490±50 to 404±48 μmol/L, both p<0.03, respectively) indicating improved glycemic control by 6% and 17%. These effects were statistically confirmed in mildly D pigs but not in overtly D pigs. In mildly D pigs, the other three cultivars of fruit showed consistent numerical but no significant improvements in glycemic control. The composition of Bitter gourd fruit was studied by metabolomics profiling and analysis identified three metabolites from the class of triterpenoids (Xuedanoside H, Acutoside A, Karaviloside IX) that were increased in the cultivars "Palee" (>3.9-fold) and "Good healthy" (>8.9-fold) compared to the mean of the other three cultivars. Bitter gourd stems and leaves from the cultivar "Bilai" increased plasma insulin concentrations in all pigs combined by 28% (from 53±6 to 67±9 pmol/L, p<0.03). The other two cultivars of stems and leaves showed consistent numerical but no significant increases in plasma insulin concentrations. The effects on plasma insulin concentrations were confirmed in mildly D pigs but not in overtly D pigs. Fruits of Bitter gourd improve glycemic control and stems-leaves of Bitter gourd increase plasma insulin concentrations in an obese pig model for mild diabetes. The effects of Bitter gourd fruit on glycemic control seem consistent but relatively small and cultivar specific which may explain the varying results of human trials reported in the literature.

The Yucatan minipig model: A new preclinical model of malnutrition in obese patients with acute or chronic diseases

Background & aimsMalnutrition can develop in patients with obesity suffering from acute or chronic illness or after obesity surgery, promoting sarcopenic obesity. A better understanding of this pathophysiology and the development of new therapeutics for chronic diseases, that are often complicated with malnutrition and obesity, justify the development of new animal experimental models close to the human physiology. This study aims to characterize the effects of obesity and underfeeding on Yucatan obese minipigs, assessing its validity as a preclinical model for obesity-related malnutrition. MethodsSixteen 30-month-old Yucatan minipigs were divided into two groups for 8 weeks: a standard diet group (ST, n=5) and an obesogenic diet group (OB, n=11). After 8 weeks, the OB group was further divided into two sub-groups: a standard diet group (OB-ST, n=5) and a low-calorie/low-protein diet group (OB-LC/LP, n=6) for 8 weeks. Body composition by CT-Scan and blood parameters were monitored, and trapezius muscle biopsies were collected to analyse signaling pathways involved in protein turnover and energy metabolism. ResultsAt W8, OB-ST animals exhibited significantly higher body weight (+37.7%, p=0.03), muscle mass (+24.9%, p=0.02), and visceral fat (+192.0%, p=0.03) compared to ST. Trapezius cross sectional area (CSA) normalized to body weight was lower in OB-ST animals (-15.02%, p=0.017). At W16, no significant changes were observed in protein turnover markers, although REDD1 increased in OB-ST (96.4%, p=0.02). After 8 weeks of low-caloric/low protein diet, OB-LC/LP showed decreased body weight (-9.8%, p=0.03), muscle mass (-6.5%, p=0.03), and visceral fat (-41.5%, p=0.03) compared to OB-ST animals. Trapezius fiber CSA significantly decreased in OB-LC/LP (-36.1%, p<0.0001) and normalized to body weight (-25.4%, p<0.0001), combined to higher ubiquitinated protein content (+38.3%, p=0.02). ConclusionOur data support that the Yucatan minipig model mimics nutritional and skeletal muscle phenotypes observed in obese patients, with or without protein-energy malnutrition. It also reproduces muscle atrophy observed in chronic diseases or post-obesity surgery, making it a promising preclinical model for obesity-related malnutrition.

Development of a functional dairy snack containing oleoylethanolamide that reduces food intake in normal-weight and obese minipigs

Oleoylethanolamide (OEA) is a safe bioactive lipid that demonstrated strong anorexigenic properties in preclinical and clinical models. In order to evaluate the importance of OEA delivery kinetic on its anorectic properties, we developed OEA-containing dairy snacks with either a liquid or a semi-solid form. The OEA+ liquid snack, but not the semi-solid one, reduced by 14 and 18 % the amount of feed eaten by normal-weight and obese minipigs, respectively, in an eating behavior test performed 4 h after snack ingestion. In vitro digestion experiments revealed that OEA release in intestinal digesta was greatly enhanced when the snack was liquid compared to the semi-solid structure. Kinetic investigations of several plasma parameters after liquid snack ingestion points towards different potential mechanism depending upon the minipig weight status, with an effect of the OEA+ liquid snack likely on endocannabinoid and other related N-acylethanolamine metabolism in normal-weight minipigs and on ketogenesis in obese ones.

Exploration of fMRI brain responses to oral sucrose after Roux-en-Y gastric bypass in obese yucatan minipigs in relationship with microbiota and metabolomics profiles

In most cases, Roux-en-Y gastric bypass (RYGBP) is an efficient intervention to lose weight, change eating behavior and improve metabolic outcomes in obese patients. We hypothesized that weight loss induced by RYGBP in obese Yucatan minipigs would induce specific modifications of the gut-brain axis and neurocognitive responses to oral sucrose stimulation in relationship with food intake control. An integrative study was performed after SHAM (n=8) or RYGBP (n=8) surgery to disentangle the physiological, metabolic and neurocognitive mechanisms of RYGBP. BOLD fMRI responses to sucrose stimulations at different concentrations, brain mRNA expression, cecal microbiota, and plasma metabolomics were explored 4 months after surgery and integrated with WGCNA analysis. We showed that weight loss induced by RYGBP or SHAM modulated differently the frontostriatal responses to oral sucrose stimulation, suggesting a different hedonic treatment and inhibitory control related to palatable food after RYGBP. The expression of brain genes involved in the serotoninergic and cannabinoid systems were impacted by RYGBP. Cecal microbiota was deeply modified and many metabolite features were differentially increased in RYGBP. Data integration with WGCNA identified interactions between key drivers of OTUs and metabolites features linked to RYGBP. This longitudinal study in the obese minipig model illustrates with a systemic and integrative analysis the mid-term consequences of RYGBP on brain mRNA expression, cecal microbiota and plasma metabolites. We confirmed the impact of RYGBP on functional brain responses related to food reward, hedonic evaluation and inhibitory control, which are key factors for the success of anti-obesity therapy and weight loss maintenance.

Despite similar clinical features metabolomics reveals distinct signatures in insulin resistant and progressively obese minipigs.

Obesity is a major contributor to the silent and progressive development of type 2 diabetes (T2D) whose prevention could be improved if individuals at risk were identified earlier. Our aim is to identify early phenotypes that precede T2D in diet-induced obese minipigs. We fed four groups of minipigs (n = 5-10) either normal-fat or high-fat high-sugar diet during 2, 4, or 6months. Morphometric features were recorded, and metabolomics and clinical parameters were assessed on fasting plasma samples. Multivariate statistical analysis on 46 morphometrical and clinical parameters allowed to differentiate 4 distinct phenotypes: NFC (control group) and three others (HF2M, HF4M, HF6M) corresponding to the different stages of the obesity progression. Compared to NFC, we observed a rapid progression of body weight and fat mass (4-, 7-, and tenfold) in obese phenotypes. Insulin resistance (IR; 2.5-fold increase of HOMA-IR) and mild dyslipidemia (1.2- and twofold increase in total cholesterol and HDL) were already present in the HF2M and remained stable in HF4M and HF6M. Plasma metabolome revealed subtle changes of 23 metabolites among the obese groups, including a progressive switch in energy metabolism from amino acids to lipids, and a transient increase in de novo lipogenesis and TCA-related metabolites in HF2M. Low anti-oxidative capacities and anti-inflammatory response metabolites were found in the HF4M, and a perturbed hexose metabolism was observed in HF6M. Overall, we show that IR and progressively obese minipigs reveal phenotype-specific metabolomic signatures for which some of the identified metabolites could be considered as potential biomarkers of early progression to TD2.

Acute Effects of Different Electroacupuncture Point Combinations to Modulate the Gut-Brain Axis in the Minipig Model.

This study aimed to compare the gut-brain axis responses to acute electroacupuncture (EA) at different acupoint combinations in the minipig model. Four adult Yucatan minipigs were subjected twice to four acute EA treatments (25-minute acute sessions) including sham (false acupoints) and control (no EA), during anesthesia and according to a Latin-square design paradigm. Acupoint combinations (4 loci each) are head-abdomen (#70 Dafengmen, #35 Sanwan), back (bilateral #27 Pishu, #28 Weishu), leg (bilateral #79 Hangou, #63 Housanli), and sham (2 bilateral points that are not acupoints). Electrocardiograms were performed to explore heart rate variability (HRV). Infrared thermography was used to measure skin temperature at the stimulation points. Saliva (cortisol) and blood samples (leptin, total/active ghrelin, insulin, and glucose) were collected for further analyses before and after acute EA. All animals were also subjected to BOLD fMRI to investigate the brain responses to EA. Acute EA significantly modulated several physiological and metabolic parameters compared to basal, sham, and/or control conditions, with contrasting effects in terms of BOLD responses in brain regions involved in the hedonic and cognitive control of food intake. The head-abdomen combination appeared to be the most promising combination in terms of brain modulation of the corticostriatal circuit, with upregulation of the dorsolateral prefrontal cortex, dorsal striatum, and anterior cingulate cortex. It also induced significantly lower plasma ghrelin levels compared to sham, suggesting anorectic effects, as well as no temperature drop at the stimulation site. This study opens the way to a further preclinical trial aimed at investigating chronic EA in obese minipigs.

Modeling microRNA-driven post-transcriptional regulation using exon-intron split analysis in pigs.

The contribution of microRNAs (miRNAs) to mRNA post-transcriptional regulation has often been explored by the post hoc selection of downregulated genes and determining whether they harbor binding sites for miRNAs of interest. This approach, however, does not discriminate whether these mRNAs are also downregulated at the transcriptional level. Here, we have characterized the transcriptional and post-transcriptional changes in mRNA expression in two porcine tissues: gluteus medius muscle of fasted and fed Duroc gilts and adipose tissue of lean and obese Duroc-Göttingen minipigs. Exon-intron split analysis of RNA-seq data allowed us to identify downregulated mRNAs with high post-transcriptional signals in fed or obese states, and we assessed whether they harbor binding sites for upregulated miRNAs in any of these two physiological states. We found 26 downregulated mRNAs with high post-transcriptional signals in the muscle of fed gilts and 21 of these were predicted targets of miRNAs upregulated in fed pigs. For adipose tissue, 44 downregulated mRNAs in obese minipigs displayed high post-transcriptional signals, and 25 of these were predicted targets of miRNAs upregulated in the obese state. These results suggest that the contribution of miRNAs to mRNA repression is more prominent in the skeletal muscle system. Finally, we identified several genes that may play relevant roles in the energy homeostasis of the pig skeletal muscle (DKK2 and PDK4) and adipose (SESN3 and ESRRG) tissues. By differentiating transcriptional from post-transcriptional changes in mRNA expression, exon-intron split analysis provides a valuable view of the regulation of gene expression, complementary to canonical differential expression analyses.

Magnesium stable isotope composition, but not concentration, responds to obesity and early insulin-resistant conditions in minipig

Hypomagnesemia is frequently associated with type 2 diabetes and generally correlates with unfavorable disease progression, but the magnesium status in pre-diabetic conditions remains unclear. Here, the magnesium metabolism is scrutinized in a minipig model of obesity and insulin resistance by measuring variations of the metallome—the set of inorganic elements—and the magnesium stable isotope composition in six organs of lean and obese minipigs raised on normal and Western-type diet, respectively. We found that metallomic variations are most generally insensitive to lean or obese phenotypes. The magnesium stable isotope composition of plasma, liver, kidney, and heart in lean minipigs are significantly heavier than in obese minipigs. For both lean and obese minipigs, the magnesium isotope composition of plasma and liver were negatively correlated to clinical phenotypes and plasma lipoproteins concentration as well as positively correlated to hyperinsulinemic-euglycemic clamp output. Because the magnesium isotope composition was not associated to insulin secretion, our results suggest that it is rather sensitive to whole body insulin sensitivity, opening perspectives to better comprehend the onset of insulin-resistant diabetic conditions.

How Does Your Body Deal With Fast-Food Meals?

Eating a less-than-healthy diet can increase the chances of becoming obese, which usually happens when body organs have difficulties using or storing the excess fat and sugar that we eat. To better understand how what we eat affects the functioning of our organs, we measured substances called metabolites, which come from food, in the blood of obese minipigs. We then used computers to figure out what happens to these metabolites inside the liver following normal meals or a fast-food meal. We found that the liver fights against unhealthy food by finding ways to use or remove the excess fat and sugar. However, some of the paths normally followed by healthy food might become blocked when we eat too much unhealthy food. Our work showed that a better understanding of how the liver processes the metabolites from unhealthy diets could help people suffering from the effects of obesity.

The effect of fatty diacid acylation of human PYY3-36 on Y2 receptor potency and half-life in minipigs

Peptides are notoriously known to display very short in vivo half-lives often measured in minutes which in many cases greatly reduces or eliminates sufficient in vivo efficacy. To obtain long half-lives allowing for up to once-weekly dosing regimen, fatty acid acylation (lipidation) have been used to non-covalently associate the peptide to serum albumin thus serving as a circulating depot. This approach is generally considered in the scientific and patent community as a standard approach to protract almost any given peptide. However, it is not trivial to prolong the half-life of peptides by lipidation and still maintain high potency and good formulation properties. Here we show that attaching a fatty acid to the obesity-drug relevant peptide PYY3-36 is not sufficient for long pharmacokinetics (PK), since the position in the backbone, but also type of fatty acid and linker strongly influences PK and potency. Furthermore, understanding the proteolytic stability of the backbone is key to obtain long half-lives by lipidation, since backbone cleavage still occurs while associated to albumin. Having identified a PYY analogue with a sufficient half-life, we show that in combination with a GLP-1 analogue, liraglutide, additional weight loss can be achieved in the obese minipig model.

A pilot study about the development and characterization of a Roux en Y gastric bypass model in obese Yucatan minipigs

Performing the Roux-en-Y gastric bypass (RYGBP) in obese Yucatan minipigs provides an opportunity to explore the mechanisms behind the effects of this surgery in controlled environmental and nutritional conditions. We hypothesized that RYGBP in these minipigs would induce changes at multiple levels, as in obese humans. We sought to characterize RYGBP in a diet-induced obese minipig model, compared with a pair-fed sham group. After inducing obesity with an ad libitum high-fat/high-sugar diet, we performed RYGBP (n = 7) or sham surgery (n = 6). Oral glucose tolerance tests (OGTT) were performed before and after surgery. Histological analyses were conducted to compare the alimentary limb at sacrifice with tissue sampled during RYGBP surgery. One death occurred in the RYGBP group at postoperative day (POD) 3. Before sacrifice, weight loss was the same across groups. GLP-1 secretion (OGTT) was significantly higher at 15, 30 and 60 min at POD 7, and at 30 and 60 min at POD 30 in the RYGBP group. Incremental insulin area under the curve increased significantly after RYGBP (p = 0.02). RYGBP induced extensive remodeling of the alimentary limb. Results show that RYGBP can be safely performed in obese minipigs, and changes mimic those observed in humans.

Determination of mitochondrial functions and damage in kidney in female LeeSung minipigs with a high-fat diet-induced obesity

The purpose of this study was to investigate the nexus between mitochondrial function and kidney injury by using a dietary-induced obese minipig model. Female Lee-Sung minipigs feeding a high-fat diet (HFD) for 6 months exhibited obesity, hyperglycaemia and dyslipidemia. HFD elevated the levels of plasma biomarkers related to renal injury, including symmetric dimethylarginine, creatinine and urea nitrogen. An extensive structural change in tubules and glomeruli was observed in HFD-fed pigs. A great amount of triacylglycerol was accumulated in HFD kidney compared to control kidney, whereas a reduction of ATP level and antioxidant capacity were exhibited in HFD kidney. Moreover, HFD altered the expressions of mitochondrial-related protein in renal cortex. To conclude, long-term HFD feeding to Lee-Sung minipigs induced obesity and kidney injury accompanied by abnormal mitochondrial functions in the renal cortex, suggesting an interrelationship with renal disease progression.

Airflow dynamics in obese minipigs with obstructive sleep apnea

ObjectivesObstructive sleep apnea (OSA) is associated with anatomical restrictions of pharyngeal airway, but the mechanism of airflow dynamics in OSA is largely unknown. This study utilized computational flow dynamics (CFD) to build a 3D model of the pharynx and to test the hypothesis that an increased restriction in the pharynx in OSA/obese minipigs leads to higher resistance, which in turn creates turbulence to induce temporary blockage of pharyngeal airway patency. DesignOf five 9-11-months-old Yucatan minipigs, 3 were non-obese (BMI<35) and two obese (BMI>51). After natural sleep monitoring using BioRadio system, pigs were sedated to collect MRI images and airflow parameters. The MRI images were processed to create 3D configurations of pharynx. These 3D configurations were meshed to create finite element models (FEM) of CFD. The obtained airflow parameters were input into the configurations to identify turbulent airflow and its location. ResultsHeavy snoring and multiple >5s hypopnea/apnea episodes (AHI = 32–35) were identified in both obese minipigs during sleep. Compared to the non-obese/non-OSA controls, obese/OSA minipigs showed much lower respiratory tidal volumes and inspiratory airflow speed. FEM simulation found that turbulence was not present in the pharynx in either model. However, a 25% increase of airflow velocity was observed at the narrowest part of the nasal pharynx in the obese/OSA minipig model. ConclusionsDespite the narrower pharyngeal airway and the higher velocity of airflow, FEM simulation indicated that turbulence was not produced in the obese/OSA minipigs.

The impact of DRP1 on myocardial fibrosis in the obese minipig.

The heart is a highly oxidative tissue, thus mitochondria play a major role in maintaining optimal cardiac function. Our previous study established a dietary-induced obese minipig with cardiac fibrosis. The aim of this study was to elucidate the role of mitochondrial dynamics in cardiac fibrosis of obese minipigs. Four-month-old Lee-Sung minipigs were randomly divided into two groups: a control group (C) and an obese group (O) by feeding a control diet or a high-fat diet (HFD) for 6months. Exposure of H9c2 cardiomyoblasts to palmitate was used to explore the effects of high-fat on induction of myocardial injury in vitro. The O pigs displayed greater heart weight and cardiac collagen accumulation. Obese pigs exhibited a lower antioxidant capacity, ATP concentration, and higher oxidative stress in the left ventricle (LV). The HFD caused downregulation in protein expression of PGC-1α and OPA1, and upregulation of DRP1, FIS1, and PINK1 in the LV of O compared to C pigs. Furthermore, palmitate induced apoptosis and decreased ATP content in H9c2 cells. Palmitate elevated the protein expression of DRP1 and PINK1 in these cells. Inhibition of DRP1 protein expression by siDRP1 in H9c2 cells resulted in enhanced ATP and decreased palmitate-induced apoptosis. These results suggest that mitochondrial dynamics were linked to the progression of obesity-related cardiac injury. Inhibition of DRP1 after palmitate exposure in H9c2 cells resulted in improved ATP level and decreased apoptosis in vitro suggesting that mitochondrial fission serves a key role in progression of obesity-induced cardiac fibrosis.

Obstructive sleep apnea in obese minipigs.

Objective:Obesity has reached epidemic proportions and is a strong risk factor for obstructive sleep apnea (OSA). However, the underlying mechanisms are poorly understood and current treatment strategies for OSA and obesity have critical limitations. Thus, establishment of an obesity-related large animal model with spontaneous OSA is imperative.Materials and methods:Natural and sedated sleep were monitored and characterized in 4 obese (body mass index - BMI>48) and 3 non-obese (BMI<40) minipigs. These minipigs were instrumented with the BioRadio system under sedation for the wireless recording of respiratory airflow, snoring, abdominal and chest respiratory movements, electroencephalogram, electrooclulogram, electromyogram, and oxygen saturation. After instrumentation, the minipigs were placed in a dark room with a remote night-vision camera for monitoring all behaviors. Wakefulness and different sleep stages were classified, and episodes of apneas and/or hypopneas were identified during natural and/or sedated sleep.Results:No hypopnea episodes were observed in two of the non-obese minipigs, but one non-obese minipig had 5 hypopnea events. Heavy snoring and 27-58 apnea and/or hypopnea episodes were identified in all 4 obese minipigs. Most of these episodes occurred in the rapid eye movement stage during natural sleep and/or sedated sleep in Yucatan minipigs.Conclusions:Obese minipigs can experience naturally occurring OSA, thus are an ideal large animal model for obese-related OSA studies.

Effects of dietary fibre and protein content on intestinal fibre degradation, short-chain fatty acid and microbiota composition in a high-fat fructose-rich diet induced obese Göttingen Minipig model.

Obesity-related metabolic syndrome has been linked with gut microbiome dysbiosis while dietary fibre (DF) and protein can modify the gut microbial ecosystem and metabolism. After 20-weeks of high-fat fructose-rich diet feeding for the development of obesity, forty-three 30-week old Göttingen Minipigs (31 ± 4.0 kg body weight) were allocated to one of the four diets with low or high DF and protein contents in a two by two factorial design and digesta were collected from the intestinal segments of minipigs after 8 weeks at libitum feeding. High DF content increased (P < 0.001) while high protein content decreased (P = 0.004) the content of non-starch polysaccharides (NSP) in all intestinal segments. Arabinoxylan (AX) as proportion of NSP was higher with high DF (P < 0.001) but decreased from the distal small intestine to the mid colon (P < 0.001). High DF increased the relative abundance of Blautia, Faecalibacterium and Peptococcus in the caecum, the mid colon and faeces, reduced the intestinal concentrations of total short-chain fatty acids (SCFA) (P = 0.020) and acetate (P = 0.011) but slightly increased butyrate pools in the large intestine (P≤ 0.050) compared to low DF. High protein increased the SCFA (P = 0.026) and propionate (P = 0.044) concentrations in the gut. High DF induced a lower increase in the BCFA concentration and proportion throughout the colon (P < 0.001). The butyrate concentrations in plasma from the jugular vein were increased with high DF diets (P = 0.031), whereas the propionate concentrations were increased (P < 0.001) and succinate were decreased (P = 0.001) with high protein diets compared with low protein diets. In conclusion, AX in the high DF diets was continuously degraded up to the mid-colon, associated with enriched butyrate-producing bacteria and slightly improved butyrate production, while protein fermentation was attenuated by high DF and high protein did not show prebiotic effects in this obese minipig model.

Preclinical Assessment of Leptin Transport into the Cerebrospinal Fluid in Diet-Induced Obese Minipigs.

ObjectiveA minipig model was employed to explore the changes in endogenous leptin transport into the central nervous system and in hypothalamic sensitivity to exogenous leptin when individuals are placed on high‐fat diet (HFD) compared with standard diet.MethodsSerum and cerebrospinal fluid (CSF) leptin concentrations during 10 weeks of HFD versus standard diet and exogenous leptin‐induced STAT3 phosphorylation in the hypothalamus of minipigs were assessed, and the hypothalamic leptin‐sensitive cells were characterized by immunofluorescence.ResultsThe efficiency of the passage of endogenous blood‐borne leptin into the CSF (measured as the log [CSF:serum leptin ratio]) decreased over time in minipigs fed a HFD (β = –0.04 ± 0.005 per kilogram of weight gain in HFD; P < 0.0001), while it remained stable in minipigs fed a standard diet. However, the ability of peripherally administered leptin to activate its receptor in hypothalamic neurons was preserved in obese minipigs at 10 weeks of HFD.ConclusionsTogether, these data are consistent with the existence of an early‐onset tranport deficiency for endogenous circulating leptin into the brain in individuals developing obesity, preceding the acquisition of hypothalamic leptin resistance. Although additional studies are required to identify the underlying mechanisms, our study paves the way for the development of new preclinical pharmacological models targeting the restoration of the shuttling of peripheral leptin into the central nervous system to manage obesity.

High‐Fat Diet Causes Mitochondrial Dysfunction and Renal Fibrosis in Minipigs

Obesity is a common and complex health problem which has been dramatically increasing in the world. It is also considered as an independent risk factor for chronic kidney disease. Kidney is one of the most energy‐demanding organs in the human body, which has the second highest mitochondrial content and oxygen consumption only next to the heart. The purpose of this study was to investigate the nexus between mitochondrial function and kidney injury by using dietary‐induced obese minipig model.Materials and methodsSix‐month‐old Lee‐Sung minipigs were assigned randomly into 2 groups and fed either a control diet (C) or a high‐fat diet (HFD) for six months, the kidney was collected for the analysis.ResultsCompared with C group, HFD group had heavier body weight, thicker back fat, higher plasma levels of glucose, triacylglycerol and total cholesterol. Furthermore, the biomarkers of kidney injury, including blood creatinine, urine creatinine, urine urea, and urine protein were higher in the HFD group than the C ones. A heavier kidney was observed in the HFD pigs; whereas a smaller ratio of kidney weight to body weight was found in the HFD group than C group, implying a less effective loading in the kidney function of HFD pig. In histologic section of kidney, disarranged structure and more glomerular fibrosis were observed in the HFD pigs. Moreover, HFD caused a decreased antioxidant capacity in the kidney, indicating an oxidative stress. Whether HFD caused the renal injury through mitochondrial mechanisms, we analyzed the ATP generation and found that HFD kidney displayed a lower ATP production than C group did. The mitochondrial‐related genes in kidney were analyzed as well. The mRNA expression of SIRT1, which has a role in variety of mitochondrial processes, including redox homeostasis and mitochondrial biogenesis was downregulated by HFD. PGC1‐1a, a nutritional sensor in the kidney, was inhibited by HFD, suggesting a decrease in mitochondrial biogenesis. UCP2, activated by mitochondrial ROS, was upregulated in HFD group compare with C group. The mRNA expressions of Opa1 and Fis1, markers of mitochondrial dynamics, were downregulated by HFD, suggesting a mitochondrial dysfunction.In conclusion, long term HFD feeding in Lee‐Sung minipigs induced obesity, dyslipidemia and renal injury. Mitochondrial dysfunction might partially contribute to the progression of obesity‐related renal damage.Support or Funding InformationThis study was sponsored by the Ministry of Science and Technology, Taiwan (MOST107‐2313‐B‐002‐049).This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Involvement of pericardial adipose tissue in cardiac fibrosis of dietary-induced obese minipigs— Role of mitochondrial function

BackgroundHeart is a high energy demand organ and cardiac fat is the main local energy source for heart. Alteration in cardiac fat may affect cardiac energy and contribute to heart dysfunction. We previously observed a link between alteration in pericardial fat (PAT) and local adverse effects on myocardial fibrosis in obese minipigs. This study investigated the role of PAT on cardiac energy and mitochondrial function, and elucidated a potential mechanism for PAT in cardiac fibrosis. Materials and methodsFive-month-old Lee-Sung minipigs were made obese by feeding a high-fat diet (HFD) for 6 months. The conditioned medium from PAT of obese minipigs (PAT-CM) was collected and H9C2 cells were treated with it to study mechanisms. ResultsHFD caused a cardiac energy deficit and fibrosis in the left ventricle. An elevated content of IL6 and malondialdehyde was found in the PAT of obese pigs. Obese pigs exhibited an increased level of oleic acid and a reduced level of saturated fatty acids in PAT compared to control pigs. HFD did not alter the metabolic characteristics of epicardial fat. PAT-CM caused apoptosis of H9C2 cells and inhibited basal mitochondrial respiration and ATP production. Protein expressions for mitochondrial dynamics- (Mfn2, Opa1, Drp1, and Fis1) and a mitophagy-related protein (Parkin) were suppressed by PAT-CM. PAT-CM enhanced the protein expression of LC3II, and the ratio of LC3II/LC3I.To conclude, PAT was involved in cardiac fibrosis of HFD-fed minipigs. The secretomes of PAT impaired mitochondrial functions and caused cardiomyocyte apoptosis in a paracrine manner.