ABSTRACT
It had been observed that homozygous albumin knockout mice (Alb-/-) exhibit low plasma free fatty acid (FFA) concentration and improved blood glucose regulation. However, it was not yet known to what extent heterozygous albumin knockout (Alb+/-) mice would display a similar phenotype. Alb-/-, Alb+/-, and wild-type (WT) female mice were studied on a low-fat diet (LFD) or high-fat diet (HFD). On both diets, decreased plasma FFA concentration, and improved glucose tolerance test were observed in Alb-/-, but not in Alb+/-, compared to WT. Plasma adiponectin concentration showed greater elevation in Alb-/- than Alb+/-. Consistent with that, adiponectin gene expression was significantly higher in Alb-/- mice than in Alb+/- and WT mice. A dose-dependent response was observed for hepatic Acadl gene expression showing higher Acadl gene expression in Alb-/- mice than in Alb+/- and WT mice. In conclusion, although female Alb+/- mice exhibited some slight differences from WT mice (e.g., increased plasma adiponectin and hepatic Acadl gene expression), Alb+/- mice did not exhibit improved glucoregulation in comparison to WT mice, indicating that a minor suppression of albumin expression is not sufficient to improve glucoregulation. Furthermore, it is now clear that although the response of female mice to HFD might be unique from how males generally respond, still the complete albumin deficiency in Alb-/- mice and the associated FFA reduction is capable of improving glucoregulation in females on this diet. The present results have implications for the role of albumin and FFA in the regulation of metabolism.
Subject(s)
Adiponectin , Albumins , Blood Glucose , Diet, High-Fat , Fatty Acids, Nonesterified , Mice, Knockout , Animals , Female , Adiponectin/genetics , Adiponectin/metabolism , Adiponectin/blood , Mice , Fatty Acids, Nonesterified/blood , Fatty Acids, Nonesterified/metabolism , Diet, High-Fat/adverse effects , Albumins/metabolism , Albumins/genetics , Blood Glucose/metabolism , Liver/metabolism , Diet, Fat-Restricted , Glucose Tolerance Test , Serum Albumin/metabolism , Serum Albumin/genetics , Gene Expression Regulation , Mice, Inbred C57BLABSTRACT
BACKGROUND: Free fatty acids (FFAs) play vital roles as energy sources and substrates in organisms; however, the molecular mechanism regulating the homeostasis of FFA levels in various circumstances, such as feeding and nonfeeding stages, is not fully clarified. Holometabolous insects digest dietary triglycerides (TAGs) during larval feeding stages and degrade stored TAGs in the fat body during metamorphosis after feeding cessation, which presents a suitable model for this study. RESULTS: This study reported that two lipases are differentially regulated by hormones to maintain the homeostasis of FFA levels during the feeding and nonfeeding stages using the lepidopteran insect cotton bollworm Helicoverpa armigera as a model. Lipase member H-A-like (Lha-like), related to human pancreatic lipase (PTL), was abundantly expressed in the midgut during the feeding stage, while the monoacylglycerol lipase ABHD12-like (Abhd12-like), related to human monoacylglycerol lipase (MGL), was abundantly expressed in the fat body during the nonfeeding stage. Lha-like was upregulated by juvenile hormone (JH) via the JH intracellular receptor methoprene-tolerant 1 (MET1), and Abhd12-like was upregulated by 20-hydroxyecdysone (20E) via forkhead box O (FOXO) transcription factor. Knockdown of Lha-like decreased FFA levels in the hemolymph and reduced TAG levels in the fat body. Moreover, lipid droplets (LDs) were small, the brain morphology was abnormal, the size of the brain was small, and the larvae showed the phenotype of delayed pupation, small pupae, and delayed tissue remodeling. Knockdown of Abhd12-like decreased FFA levels in the hemolymph; however, TAG levels increased in the fat body, and LDs remained large. The development of the brain was arrested at the larval stage, and the larvae showed a delayed pupation phenotype and delayed tissue remodeling. CONCLUSIONS: The differential regulation of lipases expression by different hormones determines FFAs homeostasis and different TAG levels in the fat body during the feeding larval growth and nonfeeding stages of metamorphosis in the insect. The homeostasis of FFAs supports insect growth, brain development, and metamorphosis.
Subject(s)
Brain , Fatty Acids, Nonesterified , Homeostasis , Animals , Brain/metabolism , Brain/growth & development , Fatty Acids, Nonesterified/metabolism , Lipase/metabolism , Lipase/genetics , Moths/growth & development , Moths/physiology , Moths/metabolism , Larva/growth & development , Larva/metabolism , Juvenile Hormones/metabolism , Insect Proteins/metabolism , Insect Proteins/genetics , Metamorphosis, Biological/physiology , Ecdysterone/metabolismABSTRACT
ATP6AP2 knockout in the renal nephron impairs receptor-mediated endocytosis, increasing urinary albumin and glucose excretion and impairing weight gain. Nonesterified fatty acids (NEFA) in urine are bound to albumin and reabsorbed in the proximal tubule through receptor-mediated endocytosis by the megalin-cubilin complex. We hypothesized that ATP6AP2 knockout increases urinary NEFA excretion through a reduction in megalin. Ten-week-old male C57BL/6 mice with nephron specific inducible ATP6AP2 knockout and noninduced controls were fed either normal diet (ND 12% fat) or high fat diet (HFD 45% fat) for 6 months. ATP6AP2 knockout significantly increased urine albumin:creatinine ratio in both ND and HFD fed mice while normalized urine NEFA concentration increased 489% and 259% in ND and HFD knockout mice compared to respective controls. Knockout decreased renal cortical megalin mRNA by 47% on ND and 49% on HFD while megalin protein expression decreased by 36% and 44% respectively. At the same time, markers of mTOR activity were increased while autophagy was impaired. Our results indicate that nephron specific ATP6AP2 knockout increases urinary NEFA excretion in the setting of impaired receptor-mediated endocytosis. Further investigation should determine whether ATP6AP2 contributes to obesity related ectopic lipid deposition in the proximal tubule.
Subject(s)
Low Density Lipoprotein Receptor-Related Protein-2 , Nephrons , Animals , Male , Mice , Diet, High-Fat , Fatty Acids/metabolism , Fatty Acids, Nonesterified/metabolism , Kidney Cortex/metabolism , Low Density Lipoprotein Receptor-Related Protein-2/genetics , Low Density Lipoprotein Receptor-Related Protein-2/metabolism , Mice, Inbred C57BL , Mice, Knockout , Nephrons/metabolism , Prorenin Receptor , Vacuolar Proton-Translocating ATPases/genetics , Vacuolar Proton-Translocating ATPases/metabolismABSTRACT
Short-chain fatty acids (SCFAs) produced in the rumen are key factors affecting dairy cows' energy balance (EB). This study aimed to quantitatively evaluate the effects of SCFAs production on EB in dairy cows. Primiparous dairy cows were divided into high non-esterified fatty acid (NEFA; group H) and low NEFA (group L) groups based on their blood NEFA levels at week 3 postpartum, which served as an indicator of EB. The amounts of SCFAs produced in the rumen, including acetate, propionate, and butyrate (SCFAsP), were calculated using the predicted rumen volume. Because there were no differences between the groups in SCFAsP/dry matter intake, whereas 4% fat-corrected milk (FCM)/SCFAsP was significantly higher in group H, it was suggested that more body fat was mobilized for milk production in group H. However, group L, which showed better EB, had propionate dominant and lower FCM/SCFAsP and milk energy/SCFAs energy at 3 and 7 weeks postpartum, indicating that group L had a better energy supply for milk production. These results suggest that SCFAsP produced by rumen fermentation and the composition of SCFAs in the rumen affect milk production and EB.
Subject(s)
Energy Metabolism , Fatty Acids, Nonesterified , Fatty Acids, Volatile , Fermentation , Lactation , Milk , Rumen , Animals , Rumen/metabolism , Cattle/metabolism , Cattle/physiology , Female , Fatty Acids, Volatile/metabolism , Lactation/metabolism , Lactation/physiology , Milk/metabolism , Fatty Acids, Nonesterified/metabolism , Fatty Acids, Nonesterified/blood , Pregnancy , Parity , Postpartum Period/metabolism , Propionates/metabolismABSTRACT
(1) Background: Proglucagon-derived peptides (PDGPs) including glucagon (Gcg), GLP-1, and GLP-2 regulate lipid metabolism in the liver, adipocytes, and intestine. However, the mechanism by which PGDPs participate in alterations in lipid metabolism induced by high-fat diet (HFD) feeding has not been elucidated. (2) Methods: Mice deficient in PGDP (GCGKO) and control mice were fed HFD for 7 days and analyzed, and differences in lipid metabolism in the liver, adipose tissue, and duodenum were investigated. (3) Results: GCGKO mice under HFD showed lower expression levels of the genes involved in free fatty acid (FFA) oxidation such as Hsl, Atgl, Cpt1a, Acox1 (p < 0.05), and Pparα (p = 0.05) mRNA in the liver than in control mice, and both FFA and triglycerides content in liver and adipose tissue weight were lower in the GCGKO mice. On the other hand, phosphorylation of hormone-sensitive lipase (HSL) in white adipose tissue did not differ between the two groups. GCGKO mice under HFD exhibited lower expression levels of Pparα and Cd36 mRNA in the duodenum as well as increased fecal cholesterol contents compared to HFD-controls. (4) Conclusions: GCGKO mice fed HFD exhibit a lesser increase in hepatic FFA and triglyceride contents and adipose tissue weight, despite reduced ß-oxidation in the liver, than in control mice. Thus, the absence of PGDP prevents dietary-induced fatty liver development due to decreased lipid uptake in the intestinal tract.
Subject(s)
CD36 Antigens , Diet, High-Fat , Intestinal Absorption , Lipid Metabolism , Liver , Mice, Knockout , PPAR alpha , Proglucagon , Animals , Male , Diet, High-Fat/adverse effects , PPAR alpha/metabolism , PPAR alpha/genetics , Liver/metabolism , Proglucagon/metabolism , Proglucagon/genetics , CD36 Antigens/metabolism , CD36 Antigens/genetics , Mice , Sterol Esterase/metabolism , Sterol Esterase/genetics , Triglycerides/metabolism , Mice, Inbred C57BL , Fatty Acids, Nonesterified/metabolism , Glucagon-Like Peptide 1/metabolism , Duodenum/metabolism , Carnitine O-Palmitoyltransferase/metabolism , Carnitine O-Palmitoyltransferase/genetics , Adipose Tissue/metabolism , Dietary Fats , Glucagon-Like Peptide 2/metabolism , Acyltransferases , LipaseABSTRACT
Recently, the incidence of NAFLD has exploded globally, but there are currently no officially approved medications for treating the condition. The regulation of NAFLD through plant-derived active substances has become a new area of interest. Quinoa (Chenopodium quinoa Willd.) has been discovered to contain a large quantity of bioactive compounds. In this study, we established a free fatty acid (FFA)-induced steatosis model and explored the effects of quinoa polyphenol extract (QPE) on the major hallmarks of NAFLD. The results indicated that QPE significantly reduced intracellular triglyceride (TG) and total cholesterol (TC) levels. Additionally, QPE remarkably elevated the levels of superoxide dismutase (SOD), catalase (CAT) and glutathione (GSH) and lowered levels of malondialdehyde (MDA). Further examination revealed that QPE attenuated intracellular inflammation, which was verified by the reduced levels of pro-inflammatory cytokines. Mechanistically, QPE inhibited fatty acid biosynthesis mainly by targeting de novo lipogenesis (DNL) via the AMPK/SREBP-1c signaling pathway. Moreover, network pharmacology was used to analyze key targets for NAFLD mitigation by ferulic acid (FA), a major component of QPE. Taken together, this study suggests that QPE could ameliorate NAFLD by modulating hepatic lipid metabolism and alleviating oxidative stress and inflammation.
Subject(s)
Chenopodium quinoa , Inflammation , Lipid Metabolism , Non-alcoholic Fatty Liver Disease , Oxidative Stress , Plant Extracts , Polyphenols , Non-alcoholic Fatty Liver Disease/drug therapy , Non-alcoholic Fatty Liver Disease/metabolism , Chenopodium quinoa/chemistry , Oxidative Stress/drug effects , Plant Extracts/pharmacology , Polyphenols/pharmacology , Lipid Metabolism/drug effects , Animals , Inflammation/drug therapy , Inflammation/metabolism , Liver/drug effects , Liver/metabolism , Male , Mice , Lipogenesis/drug effects , Humans , Mice, Inbred C57BL , Triglycerides/metabolism , Signal Transduction/drug effects , Cholesterol/metabolism , Fatty Acids, Nonesterified/metabolism , Disease Models, AnimalABSTRACT
The metabolic network's primary sources of free fatty acids (FFAs) are long- and medium-chain fatty acids of triglyceride origin and short-chain fatty acids produced by intestinal microorganisms through dietary fibre fermentation. Recent studies have demonstrated that FFAs not only serve as an energy source for the body's metabolism but also participate in regulating arterial function. Excess FFAs have been shown to lead to endothelial dysfunction, vascular hypertrophy, and vessel wall stiffness, which are important triggers of arterial hypertension and atherosclerosis. Nevertheless, free fatty acid receptors (FFARs) are involved in the regulation of arterial functions, including the proliferation, differentiation, migration, apoptosis, inflammation, and angiogenesis of vascular endothelial cells (VECs) and vascular smooth muscle cells (VSMCs). They actively regulate hypertension, endothelial dysfunction, and atherosclerosis. The objective of this review is to examine the roles and heterogeneity of FFAs and FFARs in the regulation of arterial function, with a view to identifying the points of intersection between their actions and providing new insights into the prevention and treatment of diseases associated with arterial dysfunction, as well as the development of targeted drugs.
Subject(s)
Arteries , Fatty Acids, Nonesterified , Humans , Animals , Fatty Acids, Nonesterified/metabolism , Arteries/metabolism , Atherosclerosis/metabolism , Atherosclerosis/pathology , Muscle, Smooth, Vascular/metabolism , Myocytes, Smooth Muscle/metabolism , Endothelial Cells/metabolismABSTRACT
The pituitary is the central endocrine gland with effects on metabolic dysfunction-associated steatotic liver disease (MASLD). However, it is not clear whether the pituitary responds to free fatty acid (FFA) toxicity, thus dysregulating hepatic lipid metabolism. Here, we demonstrate that decreased prolactin (PRL) levels are involved in the association between FFA and MASLD based on a liver biospecimen-based cohort. Moreover, overloaded FFAs decrease serum PRL levels, thus promoting liver steatosis in mice with both dynamic diet intervention and stereotactic pituitary FFA injection. Mechanistic studies show that excessive FFA sensing in pituitary lactotrophs inhibits the synthesis and secretion of PRL in a cell-autonomous manner. Notably, inhibiting excessive lipid uptake using pituitary stereotaxic virus injection or a specific drug delivery system effectively ameliorates hepatic lipid accumulation by improving PRL levels. Targeted inhibition of pituitary FFA sensing may be a potential therapeutic target for liver steatosis.
Subject(s)
Fatty Acids, Nonesterified , Fatty Liver , Lactotrophs , Prolactin , Animals , Prolactin/metabolism , Prolactin/blood , Fatty Acids, Nonesterified/blood , Fatty Acids, Nonesterified/metabolism , Fatty Liver/metabolism , Fatty Liver/pathology , Mice , Lactotrophs/metabolism , Lactotrophs/drug effects , Mice, Inbred C57BL , Humans , Male , Lipid Metabolism , Liver/metabolismABSTRACT
Plasma nonesterified fatty acids (NEFA) are elevated in cancer, because of decreased albumin levels and of fatty acid oxidation, and increased fatty acid synthesis and lipolysis. Albumin depletion and NEFA elevation maximally release albumin-bound tryptophan (Trp) and increase its flux down the kynurenine pathway, leading to increased production of proinflammatory kynurenine metabolites, which tumors use to undermine T-cell function and achieve immune escape. Activation of the aryl hydrocarbon receptor by kynurenic acid promotes extrahepatic Trp degradation by indoleamine 2,3-dioxygenase and leads to upregulation of poly (ADP-ribose) polymerase, activation of which and also of SIRT1 (silent mating type information regulation 2 homolog 1) could lead to depletion of NAD+ and ATP, resulting in cell death. NEFA also modulate heme synthesis and degradation, changes in which impact homocysteine metabolism and production of reduced glutathione and hydrogen sulphide. The significance of the interactions between heme and homocysteine metabolism in cancer biology has received little attention. Targeting Trp disposition in cancer to prevent the NEFA effects is suggested.
Subject(s)
Fatty Acids, Nonesterified , Neoplasms , Tryptophan , Humans , Neoplasms/metabolism , Tryptophan/metabolism , Fatty Acids, Nonesterified/metabolism , Animals , Receptors, Aryl Hydrocarbon/metabolism , Kynurenine/metabolism , Heme/metabolism , Indoleamine-Pyrrole 2,3,-Dioxygenase/metabolism , Homocysteine/metabolismABSTRACT
Intestinal disease is one of the earliest manifestations of cystic fibrosis (CF) in children and is closely tied to deficits in growth and nutrition, both of which are directly linked to future mortality. Patients are treated aggressively with pancreatic enzyme replacement therapy and a high-fat diet to circumvent fat malabsorption, but this does not reverse growth and nutritional defects. We hypothesized that defects in chylomicron production could explain why CF body weights and nutrition are so resistant to clinical treatments. We used gold standard intestinal lipid absorption and metabolism approaches, including mouse mesenteric lymph cannulation, in vivo chylomicron secretion kinetics, transmission electron microscopy, small intestinal organoids, and chylomicron metabolism assays to test this hypothesis. In mice expressing the G542X mutation in cystic fibrosis transmembrane conductance regulator (CFTR-/- mice), we find that defective FFA trafficking across the epithelium into enterocytes drives a chylomicron formation defect. Furthermore, G542X mice secrete small, triglyceride-poor chylomicrons into the lymph and blood. These defective chylomicrons are cleared into extraintestinal tissues at â¼10-fold faster than WT chylomicrons. This defect in FFA absorption resulting in dysfunctional chylomicrons cannot be explained by steatorrhea or pancreatic insufficiency and is maintained in primary small intestinal organoids treated with micellar lipids. These studies suggest that the ultrahigh-fat diet that most people with CF are counselled to follow may instead make steatorrhea and malabsorption defects worse by overloading the absorptive capacity of the CF small intestine.
Subject(s)
Chylomicrons , Cystic Fibrosis , Cystic Fibrosis/metabolism , Cystic Fibrosis/pathology , Cystic Fibrosis/genetics , Animals , Chylomicrons/metabolism , Mice , Fatty Acids, Nonesterified/metabolism , Cystic Fibrosis Transmembrane Conductance Regulator/metabolism , Cystic Fibrosis Transmembrane Conductance Regulator/genetics , Cystic Fibrosis Transmembrane Conductance Regulator/deficiency , Exocrine Pancreatic Insufficiency/metabolism , Exocrine Pancreatic Insufficiency/genetics , Exocrine Pancreatic Insufficiency/pathology , Biological Transport , Humans , Intestinal Mucosa/metabolismABSTRACT
OBJECTIVE: Obesity-induced kidney injury contributes to the development of diabetic nephropathy (DN). Here, we identified the functions of ubiquitin-specific peptidase 19 (USP19) in HK-2 cells exposed to a combination of high glucose (HG) and free fatty acid (FFA) and determined its association with TGF-beta-activated kinase 1 (TAK1). METHODS: HK-2 cells were exposed to a combination of HG and FFA. USP19 mRNA expression was detected by quantitative RT-PCR (qRT-PCR), and protein analysis was performed by immunoblotting (IB). Cell growth was assessed by Cell Counting Kit-8 (CCK-8) viability and 5-ethynyl-2'-deoxyuridine (EdU) proliferation assays. Cell cycle distribution and apoptosis were detected by flow cytometry. The USP19/TAK1 interaction and ubiquitinated TAK1 levels were assayed by coimmunoprecipitation (Co-IP) assays and IB. RESULTS: In HG+FFA-challenged HK-2 cells, USP19 was highly expressed. USP19 knockdown attenuated HG+FFA-triggered growth inhibition and apoptosis promotion in HK-2 cells. Moreover, USP19 knockdown alleviated HG+FFA-mediated PTEN-induced putative kinase 1 (PINK1)/Parkin pathway inactivation and increased mitochondrial reactive oxygen species (ROS) generation in HK-2 cells. Mechanistically, USP19 stabilized the TAK1 protein through deubiquitination. Importantly, increased TAK1 expression reversed the USP19 knockdown-mediated phenotypic changes and PINK1/Parkin pathway activation in HG+FFA-challenged HK-2 cells. CONCLUSION: The findings revealed that USP19 plays a crucial role in promoting HK-2 cell dysfunction induced by combined stimulation with HG and FFAs by stabilizing TAK1, providing a potential therapeutic strategy for combating DN.
Subject(s)
Apoptosis , Glucose , MAP Kinase Kinase Kinases , Humans , MAP Kinase Kinase Kinases/metabolism , MAP Kinase Kinase Kinases/genetics , Glucose/pharmacology , Apoptosis/drug effects , Cell Line , Fatty Acids, Nonesterified/metabolism , Fatty Acids, Nonesterified/pharmacology , Fatty Acids, Nonesterified/adverse effects , Cell Proliferation/drug effects , Ubiquitination/drug effects , Diabetic Nephropathies/metabolism , Diabetic Nephropathies/genetics , Endopeptidases/metabolism , Endopeptidases/genetics , Protein KinasesABSTRACT
Colorectal cancer (CRC) is one of the leading causes of cancer-related death. Currently, dietary factors are being emphasized in the pathogenesis of CRC. There is strong evidence that fatty acids (FAs) and free FA receptors (FFARs) are involved in CRC. This comprehensive review discusses the role of FAs and their receptors in CRC pathophysiology, development, and treatment. In particular, butyrate and n-3 polyunsaturated fatty acids have been found to exert anticancer properties by, among others, inhibiting proliferation and metastasis and inducing apoptosis in tumor cells. Consequently, they are used in conjunction with conventional therapies. Furthermore, FFAR gene expression is down-regulated in CRC, suggesting their suppressive character. Recent studies showed that the FFAR4 agonist, GW9508, can inhibit tumor growth. In conclusion, natural as well as synthetic FFAR ligands are considered promising candidates for CRC therapy.
Subject(s)
Colorectal Neoplasms , Fatty Acids, Nonesterified , Fatty Acids, Omega-3 , Receptors, G-Protein-Coupled , Humans , Colorectal Neoplasms/drug therapy , Colorectal Neoplasms/metabolism , Receptors, G-Protein-Coupled/metabolism , Fatty Acids, Nonesterified/metabolism , Fatty Acids, Omega-3/therapeutic use , Fatty Acids, Omega-3/pharmacology , Butyrates/therapeutic use , Butyrates/pharmacology , Animals , Antineoplastic Agents/therapeutic use , Antineoplastic Agents/pharmacology , Methylamines/metabolism , Apoptosis/drug effects , Cell Proliferation/drug effects , PropionatesABSTRACT
Spontaneous lipolysis results in the breakdown of milk fat by the lipoprotein lipase (EC: 3.1.1.34), an enzyme present in milk. Free fatty acids (FFAs) and by-products released in milk during lipolysis can alter both the organoleptic value of milk (off-flavors release) and technological properties of dairy products (decrease in creaming capabilities). Current climate change is having significant impacts on the feeding of grazing animals, with negative consequences on the availability and quality of grass. We and others have demonstrated that dietary restriction increases milk lipolysis in the cow species. However, no data about the impact of feed restriction on milk lipolysis is available in the ewe species. Thus, this paper aims to investigate the effect of feed restriction on milk characteristics with regard to lipolysis values in dairy ewes. Two groups of 24 multiparous Lacaune ewes in mid-lactation received a "non-restricted" control diet (100% of ad libitum DM intake) or a "restricted" (RESTR) diet (65% of ad libitum DM intake) according to a 2 × 2 crossover design. Milk gross composition together with lipolysis analyses were performed. Blood samples were also screened for metabolites or hormone concentrations. The RESTR treatment induced a decrease in milk production (- 21% compared with control treatment) and a modification of the metabolism of dairy ewes characterized by an increase in plasma non-esterified fatty acids (NEFAs), which represents the balance between adipose tissue mobilization and the use of NEFA by other tissues (+153%), cholesterol (+17%) and ß-hydroxybutyrate (+4 %) levels. As a result, a decrease in BW of dairy ewes was observed (-7%). Feed restriction also resulted in a decrease in milk lipolysis estimated by the milk FFA measured by the copper-soap method (-63 and -62%, respectively, for morning and evening milking) or by the reference Bureau of Dairy Industry method (-51 and -57%, respectively, for morning and evening milking). The decrease in milk spontaneous lipolysis under feed restriction was not associated with a decrease in lipoprotein lipase activity in ewes. These results will be completed with proteomic and lipidomic studies in milk samples to better understand mechanisms initiated in the ewe species specifically with regard to lipolysis in milk.
Subject(s)
Animal Feed , Fatty Acids, Nonesterified , Lactation , Lipolysis , Milk , Animals , Milk/chemistry , Milk/metabolism , Female , Lactation/physiology , Sheep/physiology , Animal Feed/analysis , Fatty Acids, Nonesterified/blood , Fatty Acids, Nonesterified/metabolism , Diet/veterinary , Dairying , Cross-Over Studies , Food Deprivation/physiologyABSTRACT
BACKGROUND: A major limitation of osteochondral allografts (OCA) is the deterioration of cartilage health associated with cell death during prolonged storage. However, little is known about the mechanisms that contribute to chondrocyte death during storage. PURPOSE/HYPOTHESIS: This study aimed to determine whether bioactive lipid metabolites accumulate in the storage media of OCA and whether they are associated with a loss of chondrocyte viability during prolonged storage. It was hypothesized that free fatty acids (FFAs) would accumulate over time in the storage media of OCA and adversely affect cartilage health during storage. STUDY DESIGN: Controlled laboratory study. METHODS: A group of 21 (n = 6-8 OCA/treatment group) fresh human hemicondylar OCA tissues and media were analyzed after 7, 28, and 68 days of prolonged cold (4°C) storage. Targeted mass spectrometry analysis was used to quantify bioactive FFAs, as well as primary (lipid hydroperoxide [ROOH]) and secondary (malondialdehyde) lipid oxidation products. Chondrocyte viability was measured using a fluorescence-based live/dead assay and confocal microscopy. RESULTS: The concentration of all targeted fatty acid metabolites in storage media was significantly increased with increased cold storage time (P < .05). ROOH was significantly higher on day 28 of cold storage. No difference in secondary ROOH products in storage media was observed. Chondrocyte viability significantly declined in both the en face and the vertical cross-sectional analysis with increased cold storage time and inversely correlated with fatty acid metabolites (P < .05). CONCLUSION: It is well established that elevated levels of certain FFAs and lipid oxidation products can alter cell function and cause cell death via lipotoxicity and other mechanisms. This work is the first to identify elevated levels of FFA metabolites and primary oxidation lipid products in the storage media from clinical OCA. The concentrations of FFA metabolites were measured at levels (>100 µM) known to induce cell death and were directly correlated with chondrocyte viability. CLINICAL RELEVANCE: These findings provide important targets for understanding why cartilage health declines during cold storage, which can be used to optimize media formulations and improve graft health.
Subject(s)
Cell Death , Chondrocytes , Humans , Chondrocytes/metabolism , Fatty Acids, Nonesterified/metabolism , Cell Survival , Allografts , Adult , Middle Aged , Male , Cartilage, Articular/metabolism , Female , Lipid MetabolismABSTRACT
This study aimed to determine whether obese men with nonalcoholic fatty liver disease (NAFLD) display differences between those with simple steatosis versus steatohepatitis (NASH) in splanchnic and hepatic FFA and VLDL-triglycerides (VLDL-TG) balances. The study involved 17 obese men with biopsy-proven NAFLD (9 with NASH and 8 with simple steatosis). We used hepatic vein catheterization in combination with [3H]palmitate and [14C]VLDL-TG tracers to measure splanchnic palmitate and VLDL-TG uptake and release rates during basal and hyperinsulinemic conditions. Indocyanine green was used to measure splanchnic plasma flow. Splanchnic palmitate uptake was similar in the two groups and significantly reduced during hyperinsulinemia (NASH: 62 (48-77) versus 38 (18-58) µmol/min; simple steatosis: 62 (46-78) versus 45 (25-65) µmol/min, mean (95% CI), basal versus clamp periods, respectively, P = 0.02 time-effect). Splanchnic palmitate release was also comparable between groups and nonsignificantly diminished during hyperinsulinemia. The percent palmitate delivered to the liver originating from visceral adipose tissue lipolysis was similar and unchanged by hyperinsulinemia. Splanchnic uptake and release of VLDL-TG were similar between groups. Hyperinsulinemia suppressed VLDL-TG release (P <0.05 time-effect) in both groups. Insulin-mediated glucose disposal was similar in the two groups (P = 0.54). Obese men with NASH and simple steatosis have similar splanchnic uptake and release of FFA and VLDL-TG and a similar proportion of FFA from visceral adipose tissue lipolysis delivered to the liver. These results demonstrate that the splanchnic balances of FFA and VLDL-TG do not differ between obese men with NASH and those with simple steatosis.
Subject(s)
Insulin , Lipoproteins, VLDL , Non-alcoholic Fatty Liver Disease , Triglycerides , Humans , Male , Lipoproteins, VLDL/metabolism , Non-alcoholic Fatty Liver Disease/metabolism , Non-alcoholic Fatty Liver Disease/pathology , Middle Aged , Triglycerides/metabolism , Triglycerides/blood , Insulin/metabolism , Fatty Acids, Nonesterified/metabolism , Fatty Acids, Nonesterified/blood , Adult , Fatty Liver/metabolism , Fatty Liver/pathology , Liver/metabolism , Obesity/metabolism , Obesity/complicationsABSTRACT
Fat and its hydrolysis products, fatty acids, are indispensable nutritional components; however, prolonged excessive fat consumption, particularly in western diets, contributes to the onset of obesity and multiple metabolic disorders. In this study, we propose a daily-ingestible hydrogel (denoted as ßC-MA hydrogel) composed of natural ß-glucan and sodium carboxymethylcellulose crosslinked by malic acid at 120 °C. This hydrogel exhibits rapid swelling performance, up to 24-fold within 1 min and 176-fold after 1 h in deionized water. It also lengthens gastric retention and increases endogenous satiety signal levels, potentially controlling appetite and reducing food intake. Furthermore, ßC-MA hydrogels that enter the small intestine can effectively inhibit fat hydrolysis and decrease triglyceride synthesis and transport. Specifically, the hydrogels inhibit the release of free fatty acids (FFAs) by approximately 50 % during digestion, influence the translocation of triglycerides and FFAs across the intestinal epithelium, and reduce the serum triglyceride levels by 22.2 %. These findings suggest that ßC-MA hydrogels could serve as a noninvasive gastrointestinal device for weight control, with the advantage of reducing food intake and restoring lipid metabolism homeostasis.
Subject(s)
Hydrogels , beta-Glucans , Hydrogels/chemistry , beta-Glucans/chemistry , beta-Glucans/pharmacology , Hydrolysis , Animals , Triglycerides/metabolism , Male , Mice , Lipid Metabolism/drug effects , Fatty Acids, Nonesterified/metabolism , Gastrointestinal Tract/metabolism , Gastrointestinal Tract/drug effects , Satiety Response/drug effects , RatsABSTRACT
Exploring the contribution of common microorganisms to spoilage is of great significance in inhibiting spoilage in lamb. This work investigated the extent of protein degradation and profile changes of free amino acids (FAAs), free fatty acids (FFAs) and volatile organic compounds (VOCs) in lamb caused by single- and co-culture of the common aerobic spoilage bacteria, P. paralactis, Ac. MN21 and S. maltophilia. Meanwhile, some key VOCs produced by the three bacteria during lamb spoilage were also screened by orthogonal partial least square discriminant analysis and difference value in VOCs content between inoculated groups and sterile group. Lamb inoculated with P. paralactis had the higher total viable counts, pH, total volatile base nitrogen and TCA-soluble peptides than those with the other two bacteria. Some FAAs and FFAs could be uniquely degraded by P. paralactis but not Ac. MN21 and S. maltophilia, such as Arg, Glu, C15:0, C18:0 and C18:1n9t. Co-culture of the three bacteria significantly promoted the overall spoilage, including bacterial growth, proteolysis and lipolysis. Key VOCs produced by P. paralactis were 2, 3-octanedione, those by Ac. MN21 were 1-octanol, octanal, hexanoic acid, 1-pentanol and hexanoic acid methyl ester, and that by S. maltophilia were hexanoic acid. The production of extensive key-VOCs was significantly and negatively correlated with C20:0, C23:0 and C18:ln9t degradation. This study can provide a basis for inhibiting common spoilage bacteria and promoting high-quality processing of fresh lamb.
Subject(s)
Acinetobacter , Coculture Techniques , Food Microbiology , Pseudomonas , Red Meat , Stenotrophomonas maltophilia , Volatile Organic Compounds , Animals , Volatile Organic Compounds/analysis , Volatile Organic Compounds/metabolism , Pseudomonas/metabolism , Pseudomonas/growth & development , Acinetobacter/growth & development , Acinetobacter/metabolism , Stenotrophomonas maltophilia/growth & development , Stenotrophomonas maltophilia/metabolism , Red Meat/microbiology , Red Meat/analysis , Sheep , Food Storage , Cold Temperature , Fatty Acids, Nonesterified/metabolism , Fatty Acids, Nonesterified/analysis , Amino Acids/metabolism , Amino Acids/analysis , Sheep, Domestic/microbiology , ProteolysisABSTRACT
Lipids play a pivotal role in the nutrition of preterm infants, acting as a primary energy source. Due to their underdeveloped gastrointestinal systems, lipid malabsorption is common, leading to insufficient energy intake and slowed growth. Therefore, it is critical to explore the reasons behind the low lipid absorption rate in formulas for preterm infants. This study utilized a simulated in intro gastrointestinal digestion model to assess the differences in lipid digestion between preterm human milk and various infant formulas. Results showed that the fatty acid release rates for formulas IF3, IF5, and IF7 were 58.90 %, 56.58 %, and 66.71 %, respectively, lower than human milk's 72.31 %. The primary free fatty acids (FFA) and 2-monoacylglycerol (2-MAG) released during digestion were C14:0, C16:0, C18:0, C18:1n-9, and C18:2n-6, in both human milk and formulas. Notably, the higher release of C16:0 in formulas may disrupt fatty acid balance, impacting lipid absorption. Further investigations are necessary to elucidate lipid absorption differences, which will inform the optimization of lipid content in preterm infant formulas.
Subject(s)
Digestion , Infant Formula , Infant, Premature , Milk, Human , Milk, Human/chemistry , Milk, Human/metabolism , Humans , Infant Formula/chemistry , Infant, Newborn , Fatty Acids/analysis , Fatty Acids/metabolism , Lipids/analysis , Fatty Acids, Nonesterified/analysis , Fatty Acids, Nonesterified/metabolism , Lipid Metabolism , Gastrointestinal Tract/metabolism , Models, Biological , Monoglycerides/metabolism , Monoglycerides/analysis , Dietary Fats/metabolism , Dietary Fats/analysisABSTRACT
Diabetic patients present increased volume and functional alterations in epicardial adipose tissue (EAT). We aimed to analyze EAT from type 2 diabetic patients and the inflammatory and cytotoxic effects induced on cardiomyocytes. Furthermore, we analyzed the cardioprotective role of apolipoprotein J (apoJ). EAT explants were obtained from nondiabetic patients (ND), diabetic patients without coronary disease (DM), and DM patients with coronary disease (DM-C) after heart surgery. Morphological characteristics and gene expression were evaluated. Explants were cultured for 24â¯h and the content of nonesterified fatty acids (NEFA) and sphingolipid species in secretomes was evaluated by lipidomic analysis. Afterwards, secretomes were added to AC16 human cardiomyocytes for 24â¯h in the presence or absence of cardioprotective molecules (apoJ and HDL). Cytokine release and apoptosis/necrosis were assessed by ELISA and flow cytometry. The EAT from the diabetic samples showed altered expression of genes related to lipid accumulation, insulin resistance, and inflammation. The secretomes from the DM samples presented an increased ratio of pro/antiatherogenic ceramide (Cer) species, while those from DM-C contained the highest concentration of saturated NEFA. DM and DM-C secretomes promoted inflammation and cytotoxicity on AC16 cardiomyocytes. Exogenous Cer16:0, Cer24:1, and palmitic acid reproduced deleterious effects in AC16 cells. These effects were attenuated by exogenous apoJ. Diabetic secretomes promoted inflammation and cytotoxicity in cardiomyocytes. This effect was exacerbated in the secretomes of the DM-C samples. The increased content of specific NEFA and ceramide species seems to play a key role in inducing such deleterious effects, which are attenuated by apoJ.
Subject(s)
Adipose Tissue , Diabetes Mellitus, Type 2 , Inflammation , Myocytes, Cardiac , Pericardium , Humans , Adipose Tissue/metabolism , Adipose Tissue/pathology , Adipose Tissue/drug effects , Myocytes, Cardiac/metabolism , Myocytes, Cardiac/drug effects , Myocytes, Cardiac/pathology , Pericardium/metabolism , Pericardium/pathology , Diabetes Mellitus, Type 2/metabolism , Inflammation/pathology , Inflammation/metabolism , Male , Female , Middle Aged , Aged , Apoptosis/drug effects , Lipid Metabolism/drug effects , Fatty Acids, Nonesterified/metabolism , Fatty Acids, Nonesterified/pharmacology , Epicardial Adipose TissueABSTRACT
A predigested product from arachidonic acid oil (ARA) and docosahexaenoic acid (DHA) oil in a 2:1 (w/w) ratio has been developed and evaluated in an in vitro digestion model. To produce this predigested lipid mixture, first, the two oils were enzymatically hydrolyzed up to 90% of free fatty acids (FFAs) were achieved. Then, these two fatty acid (FA) mixtures were mixed in a 2:1 ARA-to-DHA ratio (w/w) and enzymatically esterified with glycerol to produce a mixture of FFAs, mono-, di-, and triacylglycerides. Different glycerol ratios and temperatures were evaluated. The best results were attained at 10 °C and a glycerol-to-FA molar ratio of 3:1. The bio-accessibility of this predigested mixture was studied in an in vitro digestion model. A total of 90% of the digestion product was found in the micellar phase, which contained 30% monoacylglycerides, more than 50% FFAs, and a very small amount of triacylglycerols (3% w/w). All these data indicate an excellent bio-accessibility of this predigested mixture.