Common Dietary Fatty Acids Research Articles

The fa(c)ts that matter: Bumble bees differentially allocate and oxidate three common fatty acids in pollen

Pollen serves as a crucial source of protein and lipids for numerous insects. Despite the importance of pollen lipids for nutrient regulation in bees, the digestibility and absorption of different fatty acids (FAs) by bees remain poorly understood. We used 13C labeled fatty acids (FAs) to investigate the absorption and allocation of three common dietary FAs in pollen by bumble bees. Palmitic acid, the most common saturated FA in pollen, was poorly absorbed, even when supplied as tripalmitate, emulsified, or mixed in vegetable oil. In contrast, the essential linoleic acid was absorbed and allocated at the highest rate among the three FAs tested. Oleic acid, a non-essential monounsaturated FA, was absorbed and oxidized at lower rates than linoleic acid. Notably, a feeding rate experiment revealed that different fatty acids did not affect the consumption rate of pollen. This results suggests that the specific FA's absorption efficiency and allocation differ in bumble bees, impacting their utilization. These findings demonstrate the importance of considering the digestibility and absorption of different FAs. Furthermore, the study highlights the influence of pollen lipid composition on the nutritional content for pollinators and raises questions about the utilization of polyunsaturated FAs in insect metabolism.

Effects of different fatty acid ligands on the host-guest interaction of astaxanthin-bovine serum albumin: Thermodynamical analysis, binding site identification, and in vivo antioxidant evaluation

The multi-ligand binding properties of proteins make the introduction of small-molecule ligands an effective approach through which to improve the biological functions of protein-based carriers. In this study, the effect of fatty acids (FAs) as functional ligands on the self-assembly of astaxanthin (ASTA) and bovine serum albumin (BSA) and the in vivo antioxidant properties of their ternary ligand complexes were investigated. As the seven common dietary FAs, lauric acid (LAA), palmitic acid (PA), stearic acid (SA), oleic acid (OA), linoleic acid (LA), arachidonic acid (AA) and docosahexaenoic acid (DHA) were selected. The results indicated that, with the exception of DHA, six of these FAs were able to change the conformation of BSA while simultaneously improving its hydrophilicity. The introduction of FA ligands did not affect the binding mode (static quenching) or the type of intermolecular forces (hydrogen bonding and van der Waals forces) between ASTA and BSA. However, the involvement of FA ligands could cause the dominant binding domain of ASTA in BSA to migrate from the initial subdomain IIIA (site II) to subdomain IIA (site I). Simultaneously, the altered binding affinity between ASTA and BSA was also observed, and the effect of unsaturated FAs (except DHA) was more pronounced than that of saturated ones. As expected, compared with the ASTA-BSA binary complexes, the ASTA-FA-BSA terpolymer performed an increased total antioxidant capacity in vivo as well as an improved antioxidant protein expression. This study provides a new strategy for the efficient delivery of carotenoids using proteins and function ligands. • Different FA ligands altered the conformation of BSA to different degrees. • The effect of different FA ligands on the hydrophilicity of BSA was variable. • The FA ligands did not affect the binding mode between astaxanthin and BSA. • The FA ligands changed the dominant binding domain of astaxanthin in BSA. • The FA ligands improved the in vivo anti-oxidative activity of astaxanthin.

Inhibition of palmitic acid induced adipogenesis by natural polyphenols in 3T3-L1 adipocytes.

Dietary free fatty acids induce preadipocyte differentiation in the presence of a hormonal cocktail in 3T3-L1 adipocytes. Plant polyphenols are curb adipocyte differentiation and protect from metabolic stress. In the present study, we examined the effects of the saturated fatty acid, palmitic acid (PA) in presence of flavonoids, chrysin (CR) and hesperidin (HD) and phenolic acid, syringic acid (SYA) and sinapic acid (SIA). Adipocytes were incubated for 10 d with 100μmol of PA along with 10-100µmol CR/HD and 100-1000µmol SYA/SIA. PA induced clonal expansion of preadipocytes, differentiation and oxidative stress in 3T3-L1 cells following 10 d of differentiation. Adipocytes treated with PA exhibited an increase of 300% in clonal population, 110% lipid and 172% reactive oxygen species accumulation. But treatment with CR, HD, SYA and SIA in the presence of PA concentration-dependent effect was observed. Concentrations of CR/HD and SYA/SIA inhibited PA-induced mRNA expression of PPARγ, C/EBPα, SREBP-1c, FAS and NOX4. Moreover, CR, HD, SYA and SIA did not exhibit toxicity in Drosophila DNA. In summary, these results suggest that dietary fatty acids act directly on adipocytes and addition of CR, HD, SYA and SIA resulted in reduction of PA-induced negative effects on 3T3-L1 adipocytes. HIGHLIGHTS: • Palmitic acid, the common dietary free fatty acid, is known to induce adipogenesis in 3T3-L1 adipocytes. • Treatment of differentiating adipocytes with flavonoids and phenolic acids reduced palmitic acid-induced clonal expansion of preadipocytes. • Phytocompounds reduced lipid accumulation and triglyceride production as well as ROS accumulation. • Thus, the phytocompounds showed effective anti-adipogenic activity even in palmitic acid challenged environment in adipocytes.

Effect-directed analysis and chemical identification of agonists of peroxisome proliferator-activated receptors in white button mushroom.

Obesity has a serious effect on human health. It relates to metabolic syndrome, including the associated disorders such as type 2 diabetes, heart disease, stroke and hyperemia. The peroxisome proliferator-activated receptors (PPARs) are important receptors to control fat metabolism in the human body. Because of the safety concerns of synthetic drugs targeting PPARs, ligands from natural sources have drawn interest. Earlier, we have found high PPAR activities in extracts from Agaricus bisporus (white button mushroom, WBM). WBM contains a wide range of candidate compounds which could be agonists of PPARs. To identify which compounds are responsible for PPAR activation by WBM extracts, we used fractionation coupled to effect-directed analysis with reporter gene assays specific for all three PPARs for purification and LC/MS-TOF and NMR for compound identification in purified active fractions. Surprisingly, we identified the relatively common dietary fatty acid, linoleic acid, as the main ligand of PPARs in WBM. Possibly, the relatively low levels of linoleic acid in WBM are sufficient and instrumental in inducing its anti-obesogenic effects, avoiding high energy intake and negative health effects associated with high levels of linoleic acid consumption. However, it could not be excluded that a minor relatively potent compound contributes towards PPAR activation, while the anti-obesity effects of WBM may be further enhanced by receptor expression modulating compounds or compounds with completely PPAR unrelated modes of action.

Near infrared aquaphotomics study on common dietary fatty acids in cow's liquid, thawed milk

This study was aimed at two objectives: 1) explore the possibility of near infrared (NIR) spectroscopy to determine the content of common dietary fatty acids (FAs) and fat content in cow's liquid milk for better individual cow's feed management; 2) using aquaphotomics obtain better understanding of interaction between water molecular structure and FAs and gain more insight into their functionality.For these purposes, 252 milk samples were collected from 9 cows during 14 weeks and FAs content measured using gas chromatography as a reference method. Applying aquaphotomics NIR spectral analysis, quantification was attempted using 1300–1850 nm region where both water and FAs absorbance bands are located. The results showed possibility of quantification for 8 common dietary FAs which was more accurate when spectral region was narrowed to 1600–1800 nm. In the external validation, good predictions (R2> 0.75, RPD>1.5), were obtained for caproic and caprylic and acceptable for capric, lauric, myristic, myristoleic, palmitoleic and oleic acid (0.55<R2<0.75, RPD>1.5). Interpretation of influential variables in regression models revealed contribution of water absorbance bands related to protonated and hydration water, which originate from the water-FAs interaction and are important for their self-organization into assemblies with different morphologies.

FATTY ACIDS AS CELL SIGNALS IN INGESTIVE BEHAVIORS

Common dietary fatty acids, including palmitic acid, stearic acid, oleic acid, and polyunsaturated fatty acids, have been studied in the context of overall dietary fat and shown to impact on several types of behaviors, most prominently cognitive behaviors and ingestive behaviors. The independent effects of these fatty acids have been less well-delineated. Several studies implicate these common fatty acids in modulation of the CNS immune/inflammatory response as a key mediator of behavioral effects. However, signaling actions of the fatty acids to regulate cell structure and neuronal or synaptic function have been identified in numerous studies, and the relevance or contribution(s) of these to ingestive behavioral outcomes represent an area for future study. Finally, fatty acids are precursors of endocannabinoids and their structural congeners. Being highly dynamic and complex, the endocannabinoid system plays a key role ingestive behavior via cellular and synaptic mechanisms, thus representing another important area for future study.

Effects of common dietary fatty acids on milk yield and concentrations of fat and fatty acids in dairy cattle

The objective of the present article was to summarise the effects of five common dietary fatty acids (C16:0, C18:0, C18:1, C18:2 and C18:3) on the major milk fat groups (&lt;C16, C16 and C18). Forty published papers were reviewed to evaluate the effect of adding free fat or oil supplements rich in C16 and C18 fatty acids on the response of milk fat secretion and composition. From those 40 studies, 21 were used to investigate the effect of total dietary concentration of C16:0, C18:0, C18:1, C18:2 and C18:3 on milk secretion or concentrations of milk &lt;C16, C16 and C18 fatty acid groups. The results indicated that C16 supplementation increased total milk fatty acids, mainly by increasing milk C16 yield, without affecting milk &lt;C16 and C18 yield. Supplements rich in unsaturated fatty acid decreased total milk fatty acid by inhibiting secretion of milk fatty acids shorter than C18, with linoleic acid being the most inhibitory. Mixtures of feed fatty acid (C16:0 + C18:0 and C16:0 + C18:1) did not significantly affect total milk fatty acid yield. According to regression of milk C16 yield on dietary fatty acid, endogenous C16 contributes ~80% of total milk C16, but this proportion varies with the level and type of dietary fatty acid fed. Milk mid-infrared analysis can be used to routinely measure the presence of milk &lt;C16 fatty acid, the concentration of which provides a good indicator of inhibition of milk fatty acid secretion. In contrast, measurement of total milk fat content is less effective as a diagnostic tool due to the masking effect of the exogenous supply of C16 and C18 dietary fatty acids.

The Modulatory Effect of Common Dietary Fatty Acids on IPEC‐J2 Transport of Lipopolysaccharide and Monolayer Barrier Integrity in vitro

Recent evidence has demonstrated the ability of dietary fatty acid composition to influence postprandial endotoxemia in humans and swine. However, the mechanism(s) by which fatty acids affect transport of lipopolysaccharide (LPS) across the intestinal epithelium has not yet been fully elucidated. To study this, we utilized non‐transformed porcine jejunum‐derived IPEC‐J2 cells. This cell line is well‐recognized to form confluent polarized monolayers with apical and basolateral regions, allowing for an in vitro model of the intestinal epithelium. The present study investigated the effect dietary fatty acids and inflammation had on intestinal permeability of LPS and barrier integrity in IPEC‐J2 monolayers. We hypothesized that treatment of IPEC‐J2 monolayer with saturated or n‐6 polyunsaturated fatty acids (PUFA) would enhance fluorescein‐isothiocyanate (FITC)‐LPS apical‐to‐basolateral permeability and reduce transepithelial resistance (TER); whereas n‐3 PUFA would attenuate this response. Secondly, we hypothesized that lauric acid would amplify the reduction in monolayer integrity associated with pre‐treatment with LPS. IPEC‐J2 cell monolayers were grown to confluency (&gt;1kΩ) on 0.4 μm Transwell® inserts. In experiment 1, these monolayers were treated with either 50 or 100 μM of docosahexaenoic acid, lauric acid, arachidonic acid or vehicle for 48 hours before TER and FITC‐LPS determination of barrier integrity. In experiment 2, monolayers were treated with either 5 μg/mL LPS, LPS plus 100 μM lauric acid or saline vehicle for 24 hours and then TER and FITC‐LPS determination of barrier integrity assessed. In experiment 1, TER and FITC‐LPS permeability was not altered by fatty acid type or concentration. For experiment 2, compared to the saline control, LPS caused a significant time‐dependent reduction in TER (p&lt;0.001); however, the co‐inclusion of lauric acid fatty acids did not have any additive effect on TER (p&gt;0.05). Although TER was reduced, surprisingly we reported no treatment differences in apical to basolateral FITC‐LPS permeability. In conclusion, fatty acid treatment did not modulate barrier inflammation and permeability in in vitro IPEC‐J2 monolayers. However, live Gram‐negative bacteria or their bacterial cellular membrane fragments may be a better model to study the effects of fatty acids on LPS permeability.Support or Funding InformationResearch supported by the Agriculture and Food Research Initiative Grant No. 2014‐67017‐21778 from the USDA NIFA.

3T3-L1 Preadipocytes Exhibit Heightened Monocyte-Chemoattractant Protein-1 Response to Acute Fatty Acid Exposure

Preadipocytes contribute to the inflammatory responses within adipose tissue. Whilst fatty acids are known to elicit an inflammatory response within adipose tissue, the relative contribution of preadipocytes and mature adipocytes to this is yet to be determined. We aimed to examine the actions of common dietary fatty acids on the acute inflammatory and adipokine response in 3T3-L1 preadipocytes and differentiated mature adipocytes. Gene expression levels of key adipokines in 3T3-L1 preadipocytes and adipocytes were determined following incubation with palmitic acid, myristic acid or oleic acid and positive inflammatory control, lipopolysaccharide for 2 and 4 h. Inflammatory kinase signalling was assessed by analysis of nuclear factor-κB, p38-mitogen-activated protein kinase and c-jun amino-terminal kinase phosphorylation. Under basal conditions, intracellular monocyte chemoattractant protein-1 and interleukin-6 gene expression levels were increased in preadipocytes, whereas mature adipocytes expressed increased gene expression levels of leptin and adiponectin. Fatty acid exposure at 2 and 4 h increased both monocyte chemoattractant protein-1 and interleukin-6 gene expression levels in preadipocytes to greater levels than in mature adipocytes. There was an accompanying increase of inhibitor of κB-α degradation and nuclear factor-κB (p65) (Ser536) phosphorylation with fatty acid exposure in the preadipocytes only. The current study points to preadipocytes rather than the adipocytes as the contributors to both immune cell recruitment and inflammatory adipokine secretion with acute increases in fatty acids.

Transient and permanent experience with fatty acids changes Drosophila melanogaster preference and fitness.

Food and host-preference relies on genetic adaptation and sensory experience. In vertebrates, experience with food-related cues during early development can change adult preference. This is also true in holometabolous insects, which undergo a drastic nervous system remodelling during their complete metamorphosis, but remains uncertain in Drosophila melanogaster. We have conditioned D. melanogaster with oleic (C18∶1) and stearic (C18∶0) acids, two common dietary fatty acids, respectively preferred by larvae and adult. Wild-type individuals exposed either during a transient period of development–from embryo to adult–or more permanently–during one to ten generation cycles–were affected by such conditioning. In particular, the oviposition preference of females exposed to each fatty acid during larval development was affected without cross-effect indicating the specificity of each substance. Permanent exposure to each fatty acid also drastically changed oviposition preference as well as major fitness traits (development duration, sex-ratio, fecundity, adult lethality). This suggests that D. melanogaster ability to adapt to new food sources is determined by its genetic and sensory plasticity both of which may explain the success of this generalist-diet species.

Induction of Autophagy by Palmitic Acid via Protein Kinase C-mediated Signaling Pathway Independent of mTOR (Mammalian Target of Rapamycin)

Lipotoxicity refers to the cytotoxic effects of excess fat accumulation in cells and has been implicated as one of the contributing factors to diseases like obesity, diabetes, and non-alcoholic fatty liver. In this study we sought to examine effects of palmitic acid (PA) and oleic acid, two of the common dietary fatty acids on the autophagic process. We found that PA, but not oleic acid, was able to cause an increase in autophagic flux, evidenced by LC3-II accumulation and formation of GFP-LC3 puncta. Notably, PA-induced autophagy was found to be independent of mTOR regulation. Next, in search of the mechanism mediating PA-induced autophagy, we found increased levels of diacylglycerol species and protein kinase C (PKC) activation in PA-treated cells. More importantly, inhibition of classical PKC isoforms (PKC-α) was able to effectively suppress PA-induced autophagy. Finally, we showed that inhibition of autophagy sensitized the cells to PA-induced apoptosis, suggesting the pro-survival function of autophagy induced by PA. Taken together, results from this study reveal a novel mechanism underlying free fatty acid-mediated autophagy. Furthermore, the pro-survival function of autophagy suggests modulation of autophagy as a potential therapeutic strategy in protection of cells against lipotoxicity and lipid-related metabolic diseases.

C&amp;lt;sub&amp;gt;16&amp;lt;/sub&amp;gt; Saturated Fatty Acid Induced Autophagy in A549 Cells through Topoisomerase I Inhibition

C16 saturated fatty acid (Palmitic acid) is one of the most common dietary fatty acids which played an important role in the cellular biological functions. Palmitic acid (PA) was tested for potential inhibition of DNA topoisomerase I (topo I) and it exhibited inhibitory activity in the nanomolar range. Treatment of lung adenocarcinoma cell line A549 with PA resulted in a decrease in cell viability in a concentration-dependent manner, and PA showed cytotoxicity with an IC50 value of 150 μM. DNA fragmentation assay and caspase activity indicated that PA does not induce apoptotic cell death in A549 cells. Finally, we found that PA was able to cause an increase in autophagic flux in a time-dependent manner, evidenced by the accumulation of LC3 through monodansylcadaverine (MDC) staining. More importantly, inhibition of autophagy by blocking autophagosome formation via the inhibition of type III Phosphatidylinositol 3-kinases (PI-3K), by 3-Methyladenine (3-MA) was able to effectively suppress PA-induced autophagy. We showed that inhibition of autophagy sensitized the cells signal to PA-induced apoptosis, suggesting the pro-survival function of autophagy induced by PA. Taken together, results from this study reveal that PA as a topo I inhibitor induced autophagic cell death in A549 cells.

Liver type fatty acid binding protein (L-FABP) gene ablation reduces nuclear ligand distribution and peroxisome proliferator-activated receptor-α activity in cultured primary hepatocytes

The effect of liver type fatty acid binding protein (L-FABP) gene ablation on the uptake and distribution of long chain fatty acids (LCFA) to the nucleus by real-time laser scanning confocal imaging and peroxisome proliferator-activated receptor-α (PPARα) activity was examined in cultured primary hepatocytes from livers wild-type L-FABP+/+ and gene ablated L-FABP−/− mice. Cultured primary hepatocytes from livers of L-FABP−/− mice exhibited: (i) reduced oxidation of palmitic acid, a common dietary long chain fatty acid (LCFA); (ii) reduced expression of fatty acid oxidative enzymes–proteins transcriptionally regulated by PPARα; (iii) reduced palmitic acid-induced PPARα co-immunoprecipitation with coactivator SRC-1 concomitant with increased PPARα co-immunoprecipitation with coinhibitor N-CoR; (iv) reduced palmitic acid-induced PPARα. Diminished PPARα activation in L-FABP null hepatocytes was associated with lower uptake of common dietary LCFA (palmitic acid as well as its fluorescent derivative BODIPY FL C 16), reduced level of total unesterified LCFA, and real-time redistribution of BODIPY FL C 16 from the central nucleoplasm to the nuclear envelope. Taken together, these studies support the hypothesis that L-FABP may facilitate ligand (LCFA)-activated PPARα transcriptional activity at least in part by increasing total LCFA ligand available to PPARα for inducing PPARα-mediated transcription of proteins involved in LCFA metabolism.

Oleate Reverses Palmitate-induced Insulin Resistance and Inflammation in Skeletal Muscle Cells

Here we report that in skeletal muscle cells the contribution to insulin resistance and inflammation of two common dietary long-chain fatty acids depends on the channeling of these lipids to distinct cellular metabolic fates. Exposure of cells to the saturated fatty acid palmitate led to enhanced diacylglycerol levels and the consequent activation of the protein kinase C/nuclear factor kappaB pathway, finally resulting in enhanced interleukin 6 secretion and down-regulation of the expression of genes involved in the control of the oxidative capacity of skeletal muscle (peroxisome proliferator-activated receptor (PPAR)gamma-coactivator 1alpha) and triglyceride synthesis (acyl-coenzyme A: diacylglycerol acyltransferase 2). In contrast, exposure to the monounsaturated fatty acid oleate did not lead to these changes. Interestingly, co-incubation of cells with palmitate and oleate reversed both inflammation and impairment of insulin signaling by channeling palmitate into triglycerides and by up-regulating the expression of genes involved in mitochondrial beta-oxidation, thus reducing its incorporation into diacylglycerol. Our findings support a model of cellular lipid metabolism in which oleate protects against palmitate-induced inflammation and insulin resistance in skeletal muscle cells by promoting triglyceride accumulation and mitochondrial beta-oxidation through PPARalpha- and protein kinase A-dependent mechanisms.

Omega-3 Fatty Acids and Cardiac Arrhythmias: Prior Studies and Recommendations for Future Research

Compared with prehistoric times, the ratio of n-6 to n-3 fatty acids in the modern diet has increased ≈10-fold to 20:1.1,2 A substantial body of evidence suggests that n-3 polyunsaturated fatty acids (PUFAs) provide cardiovascular protection and prevent arrhythmias.3–5 This has led to the recommendation by the American Heart Association that all adults eat fatty fish at least 2 times per week and that patients with coronary heart disease (CHD) are advised to consume ≈1 g/d of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) combined.6,7 The evidence base is not entirely consistent, and a number of randomized trials have failed to show a protective effect of n-3 PUFAs against arrhythmias.8–10 This has led to some uncertainty regarding the appropriate recommendations for their use.11 The present review originates from the Omega-3 Fatty Acids and Their Role in Cardiac Arrhythmogenesis Workshop sponsored by the National Heart, Lung, and Blood Institute and the Office of Dietary Supplements on August 29–30, 2005, and includes the findings from the recently published trials. Data from epidemiological studies, randomized clinical trials, animal studies, and basic science mechanistic studies on the role of n-3 PUFAs in arrhythmia prevention are examined. Areas in which the data are conflicting or our current knowledge is lacking are emphasized. Fatty acids are classified by the length of the carbon chain (long chain, n=20 to 22; intermediate chain, n=18) and the number of double bonds (saturated, monounsaturated, polyunsaturated).1,2 For PUFAs, the location of the first double bond relative to the -CH3 or omega (n-) end is given. Long- and intermediate-chain fatty acids must be ingested as part of the diet because they cannot be synthesized by humans and are therefore referred to as essential. The most common dietary fatty acids include (1) the omega-6 linoleic acid …

A Model for Fatty Acid Transport into the Brain

A key function of fatty acid (FA) transport into the brain is to supply polyunsaturated fatty acids (PUFA) that are not synthesized in brain cells but are essential signaling molecules and components of the phospholipid membrane. In addition, common dietary FAs such as palmitic acid are also rapidly taken up by the brain and esterified to phospholipids or oxidized to provide cellular energy. Most evidence shows that FA crossing the blood brain barrier (BBB) is derived mainly from FA/albumin complexes and, to a lesser extent, from circulating lipoproteins. Our model proposes that FA diffuse across the lipid bilayer of the BBB without specific transporters to reach brain cells. They cross the luminal and transluminal leaflets of the endothelial cells and the plasma membrane of neural cells by reversible flip-flop. Acyl-CoA synthetases trap FA by forming acyl-CoA, which cannot diffuse out of the cell. Selection of FA is controlled largely by enzymes in the pathways of intracellular metabolism, beginning with the acyl-CoA synthetase.

Liver Fatty Acid-binding Protein Gene Ablation Inhibits Branched-chain Fatty Acid Metabolism in Cultured Primary Hepatocytes

Whereas the role of liver fatty acid-binding protein (L-FABP) in the uptake, transport, mitochondrial oxidation, and esterification of normal straight-chain fatty acids has been studied extensively, almost nothing is known regarding the function of L-FABP in peroxisomal oxidation and metabolism of branched-chain fatty acids. Therefore, phytanic acid (most common dietary branched-chain fatty acid) was chosen to address these issues in cultured primary hepatocytes isolated from livers of L-FABP gene-ablated (-/-) and wild type (+/+) mice. These studies provided three new insights: First, L-FABP gene ablation reduced maximal, but not initial, uptake of phytanic acid 3.2-fold. Initial uptake of phytanic acid uptake was unaltered apparently due to concomitant 5.3-, 1.6-, and 1.4-fold up-regulation of plasma membrane fatty acid transporter/translocase proteins (glutamic-oxaloacetic transaminase, fatty acid transport protein, and fatty acid translocase, respectively). Second, L-FABP gene ablation inhibited phytanic acid peroxisomal oxidation and microsomal esterification. These effects were consistent with reduced cytoplasmic fatty acid transport as evidenced by multiphoton fluorescence photobleaching recovery, where L-FABP gene ablation reduced the cytoplasmic, but not membrane, diffusional component of NBD-stearic acid movement 2-fold. Third, lipid analysis of the L-FABP gene-ablated hepatocytes revealed an altered fatty acid phenotype. Free fatty acid and triglyceride levels were decreased 1.9- and 1.6-fold, respectively. In summary, results with cultured primary hepatocytes isolated from L-FABP (+/+) and L-FABP (-/-) mice demonstrated for the first time a physiological role of L-FABP in the uptake and metabolism of branched-chain fatty acids.

Triglyceride accumulation protects against fatty acid-induced lipotoxicity.

Excess lipid accumulation in non-adipose tissues is associated with insulin resistance, pancreatic beta-cell apoptosis and heart failure. Here, we demonstrate in cultured cells that the relative toxicity of two common dietary long chain fatty acids is related to channeling of these lipids to distinct cellular metabolic fates. Oleic acid supplementation leads to triglyceride accumulation and is well tolerated, whereas excess palmitic acid is poorly incorporated into triglyceride and causes apoptosis. Unsaturated fatty acids rescue palmitate-induced apoptosis by channeling palmitate into triglyceride pools and away from pathways leading to apoptosis. Moreover, in the setting of impaired triglyceride synthesis, oleate induces lipotoxicity. Our findings support a model of cellular lipid metabolism in which unsaturated fatty acids serve a protective function against lipotoxicity though promotion of triglyceride accumulation.

Oleic acid-induced mucosal injury in developing piglet intestine.

A role for luminal nutrients, in particular products of lipid digestion, in the pathogenesis of mucosal injury to developing intestine has been postulated. We evaluated changes in mucosal permeability and light and electron microscopic histology induced by luminal perfusion with the long-chain fatty acid oleate in developing piglet intestine as a function of age and concentration of the fatty acid. 51Cr-labeled EDTA plasma-to-lumen clearance was measured in jejunum and ileum of 1-day-, 3-day-, 2-wk-, and 1-mo-old piglets during sequential perfusion with saline control (20 min); 0, 1, 5, and 10 mM oleic acid/10 mM taurocholate in saline (20 min); and normal saline (60 min). The jejunum of piglets < or = 2 wk showed significantly greater increases in mucosal permeability compared with 1-mo-old animals after perfusion with oleic acid. This effect was dependent on the luminal concentration of the fatty acid and was associated with mucosal injury evident under light and electron microscopy. In contrast, the overall response in ileum was more attenuated compared with jejunum. Thus oleic acid, a common dietary fatty acid, induces dose- and age-dependent injury in developing piglet intestine. Investigation of the mechanisms of this injury may provide the basis for dietary modifications directed at decreasing the risk of mucosal injury during enteral feeding in neonatal intestine.

Incorporation of arachidonic, dihomogamma linolenic and eicosapentaenoic acids into cultured V79 cells

The uptake and distribution of three common dietary polyunsaturated fatty acids was studied using Chinese hamster lung fibroblasts (V79 cells). Treatment of V79 cells with arachidonic (20:4), eicosapentaenoic (20:5) and dihomogammalinolenic (20.3) acids for 24 hr produced a marked uptake of 20:3 and 20:4, both of which were assimilated to a considerably greater degree than 20:5. All polyunsaturated fatty acids were incorporated primarily into phospholipids; however, there were considerable differences in their distribution into individual phospholipid species. Although 20:4 was incorporated primarily into phosphatidylcholine, 20:3 entered largely into phosphatidylethanolamine and phosphatidylglycerol, and 20:5 was distributed about equally between phosphatidylcholine, phosphatidylethanolamine and phosphatidylinositol. A marked conversion of 20:3 to 20:4 was found after 24 hr and, in several phospholipids, there was as much derived-radiolabeled 20:4 as there was radiolabeled 20:3. There was little evidence of 20:4 and 20:5 metabolism. V79 cells undergo substantial changes in phospholipid fatty acid composition following supplementation with these polyunsaturated fatty acids; however, these fatty acids are assimilated to different degrees and their distribution among cellular phospholipids is distinct, suggesting incorporation via independent mechanisms.