RESUMO
A key factor limiting the study of the origin and metabolism of brain fatty acids is the lack of cost-efficient methods available to trace fatty acids. Here, through the application of compound-specific isotope analysis (CSIA), a novel, cost-efficient method, we successfully differentiated between brain DHA originating directly from dietary omega (n)-3 polyunsaturated fatty acids (PUFA), and brain DHA biochemically synthesized to determine the origin of brain DHA in fat-1 mice. Fat-1 mice and their wild-type littermates were either weaned onto n-6 PUFA rich, n-3 PUFA deficient diets or diets rich in both n-3 and n-6 PUFA. Isotopic analysis of fatty acid methyl esters from brain and liver tissue was conducted via gas chromatography- isotope ratio mass spectrometry. Our data demonstrates that in the presence of n-3 and n-6 PUFA, fat-1 mice obtain their brain DHA solely from n-3 PUFA sources. This study reflects the first application of CSIA to a complex multivariate model to determine the origin of brain fatty acids.
Assuntos
Encéfalo/metabolismo , Isótopos de Carbono/análise , Gorduras na Dieta/administração & dosagem , Ácidos Docosa-Hexaenoicos/análise , Animais , Ácidos Docosa-Hexaenoicos/biossíntese , Ácidos Graxos Ômega-3/administração & dosagem , Ácidos Graxos Ômega-6/administração & dosagem , Feminino , Cromatografia Gasosa-Espectrometria de Massas , Fígado/metabolismo , Masculino , CamundongosRESUMO
Saturated fatty acids are the most abundant fatty acids in the brain, however, there has been some debate regarding the ability of intact dietary saturated fatty acids to be incorporated into the brain. In the present study, we use compound specific isotope analysis to measure the natural abundance carbon isotopic signature of brain, liver, and blood palmitic acid (PAM) and compare it to the dietary PAM and sugar isotopic signatures to calculate the relative contribution of both the incorporation of intact and endogenously synthesized PAM to these pools. Mice were equilibrated to the study diet, and extracted fatty acids were analyzed with gas chromatography isotope ratio mass spectrometry to determine the carbon isotopic signature of PAM (δ13 CPAM ). Liver, serum total, and serum unesterified fatty acid δ13 CPAM ranged between -20.6 and -21.1 mUr and were approximately 8.5 mUr more enriched in 13 C when compared to the dietary PAM signature. Brain δ13 CPAM was found to be more enriched than liver or blood pools (-16.7 ± 0.2 mUr, mean ± SD). Two end-member-mixed modeling using the carbon isotopic signature of dietary PAM and dietary sugars determined the contribution of synthesis to the total tissue PAM pool to range between 44% and 48%. This suggests that endogenous synthesis and dietary PAM are near equal contributors to brain, liver, and blood PAM pools. In conclusion, our data provide evidence that brain PAM levels are maintained by both local endogenous synthesis and through the uptake of intact PAM from the blood.
Assuntos
Encéfalo/metabolismo , Ácido Palmítico/sangue , Animais , Isótopos de Carbono , Dieta , Ácidos Graxos Ômega-3/administração & dosagem , Feminino , Camundongos , Camundongos Endogâmicos BALB C , GravidezRESUMO
Resolution of inflammation in the periphery was once thought to be a passive process, but new research now suggests it is an active process mediated by specialized pro-resolving lipid mediators (SPM) derived from omega-3 polyunsaturated fatty acids (n-3 PUFA). However, this has yet to be illustrated in neuroinflammation. The purpose of this study was to measure resolution of neuroinflammation and to test whether increasing brain docosahexaenoic acid (DHA) affects the resolution of neuroinflammation. C57Bl/6 mice, fat-1 mice and their wildtype littermates, fed either fish oil or safflower oil, received lipopolysaccharide (LPS) in the left lateral ventricle. Animals were then euthanized at various time points for immunohistochemistry, gene expression, and lipidomic analyses. Peak microglial activation was observed at 5 days post-surgery and the resolution index was 10 days. Of the approximately 350 genes significantly changed over the 28 days post LPS injection, 130 were uniquely changed at 3 days post injection. No changes were observed in the bioactive mediator pools. However, a few lysophospholipid species were decreased at 24hr post surgery. When brain DHA is increased, microglial cell density did not resolve faster and did not alter gene expression. In conclusion, resolution of neuroinflammation appears to be independent of SPM. Increasing brain DHA had no effect in this model.
Assuntos
Encéfalo/metabolismo , Ácidos Docosa-Hexaenoicos/metabolismo , Óleos de Peixe/administração & dosagem , Lipopolissacarídeos/toxicidade , Óleo de Cártamo/administração & dosagem , Animais , Encéfalo/efeitos dos fármacos , Inflamação/induzido quimicamente , Inflamação/dietoterapia , Inflamação/metabolismo , Injeções Intraventriculares , Lipopolissacarídeos/administração & dosagem , Masculino , Aprendizagem em Labirinto/efeitos dos fármacos , Aprendizagem em Labirinto/fisiologia , Camundongos , Camundongos Endogâmicos C3H , Camundongos Endogâmicos C57BLRESUMO
DHA (22:6n-3) may be derived from two dietary sources, preformed dietary DHA or through synthesis from α-linolenic acid (ALA; 18:3n-3). However, conventional methods cannot distinguish between DHA derived from either source without the use of costly labeled tracers. In the present study, we demonstrate the proof-of-concept that compound-specific isotope analysis (CSIA) by GC-isotope ratio mass spectrometry (IRMS) can differentiate between sources of brain DHA based on differences in natural 13C enrichment. Mice were fed diets containing either purified ALA or DHA as the sole n-3 PUFA. Extracted lipids were analyzed by CSIA for natural abundance 13C enrichment. Brain DHA from DHA-fed mice was significantly more enriched (-23.32 to -21.92) compared with mice on the ALA diet (-28.25 to -27.49). The measured 13C enrichment of brain DHA closely resembled the dietary n-3 PUFA source, -21.86 and -28.22 for DHA and ALA, respectively. The dietary effect on DHA 13C enrichment was similar in liver and blood fractions. Our results demonstrate the effectiveness of CSIA, at natural 13C enrichment, to differentiate between the incorporation of preformed or synthesized DHA into the brain and other tissues without the need for tracers.
Assuntos
Encéfalo/metabolismo , Gorduras na Dieta/metabolismo , Ácidos Docosa-Hexaenoicos/química , Ácidos Docosa-Hexaenoicos/metabolismo , Animais , Feminino , Isótopos , Masculino , Metilação , Camundongos , Camundongos Endogâmicos BALB C , GravidezRESUMO
BACKGROUND: Neuroinflammation is a proposed mechanism by which Alzheimer's disease (AD) pathology potentiates neuronal death and cognitive decline. Consumption of omega-3 polyunsaturated fatty acids (PUFA) is associated with a decreased risk of AD in human observational studies and exerts protective effects on cognition and pathology in animal models. These fatty acids and molecules derived from them are known to have anti-inflammatory and pro-resolving properties, presenting a potential mechanism for these protective effects. METHODS: Here, we explore this mechanism using fat-1 transgenic mice and their wild type littermates weaned onto either a fish oil diet (high in n-3 PUFA) or a safflower oil diet (negligible n-3 PUFA). The fat-1 mouse carries a transgene that enables it to convert omega-6 to omega-3 PUFA. At 12 weeks of age, mice underwent intracerebroventricular (icv) infusion of amyloid-ß 1-40. Brains were collected between 1 and 28 days post-icv, and hippocampal microglia, astrocytes, and degenerating neurons were quantified by immunohistochemistry with epifluorescence microscopy, while microglia morphology was assessed with confocal microscopy and skeleton analysis. RESULTS: Fat-1 mice fed with the safflower oil diet and wild type mice fed with the fish oil diet had higher brain DHA in comparison with the wild type mice fed with the safflower oil diet. Relative to the wild type mice fed with the safflower oil diet, fat-1 mice exhibited a lower peak in the number of labelled microglia, wild type mice fed with fish oil had fewer degenerating neurons, and both exhibited alterations in microglia morphology at 10 days post-surgery. There were no differences in astrocyte number at any time point and no differences in the time course of microglia or astrocyte activation following infusion of amyloid-ß 1-40. CONCLUSIONS: Increasing brain DHA, through either dietary or transgenic means, decreases some elements of the inflammatory response to amyloid-ß in a mouse model of AD. This supports the hypothesis that omega-3 PUFA may be protective against AD by modulating the immune response to amyloid-ß.