RESUMO
Polyunsaturated fatty acids and their metabolites have been reported in which their pathway has potential for the modulation of cancer cell growth. 13-(S)-HODE and 15-(S)-HETE, both of which are main metabolites of 15-LOXs, play an important role as endogenous ligands in biological systems. However, the modification of 13-(S)-HODE and 15-(S)-HETE in pharmaceutical applications has not been explored widely. Herein, we report the stereoselective syntheses of 13-(S)-HODE, 15-(S)-HETE, and its derivatives to enable the synthesis of bioactive fatty acid analogues.
Assuntos
Ácidos Graxos Insaturados , Ácidos Hidroxieicosatetraenoicos , Estereoisomerismo , Ácidos Hidroxieicosatetraenoicos/química , Ácidos Hidroxieicosatetraenoicos/síntese química , Ácidos Graxos Insaturados/química , Ácidos Graxos Insaturados/síntese química , Estrutura Molecular , Ácidos Linoleicos/química , Ácidos Linoleicos/síntese químicaRESUMO
We propose a co-immobilized chemo-enzyme cascade system to mitigate random intermediate diffusion from the mixture of individual immobilized catalysts and achieve a one-pot reaction of multi-enzyme and reductant. Catalyzed by lipase and lipoxygenase, unsaturated lipid hydroperoxides (HPOs) were synthesized. 13(S)-hydroperoxy-9Z, 11E-octadecadienoic acid (13-HPODE), one compound of HPOs, was subsequently reduced to 13(S)-hydroxy-9Z, 11E-octadecadienoic acid (13-HODE) by cysteine. Upon the optimized conditions, 75.28 mg of 13-HPODE and 4.01 mg of 13-HODE were produced from per milliliter of oil. The co-immobilized catalysts exhibited improved yield compared to the mixture of individually immobilized catalysts. Moreover, it demonstrated satisfactory durability and recyclability, maintaining a relative HPOs yield of 78.5% after 5 cycles. This work has achieved the co-immobilization of lipase, lipoxygenase and the reductant cysteine for the first time, successfully applying it to the conversion of soybean oil into 13-HODE. It offers a technological platform for transforming various oils into high-value products.
Assuntos
Cisteína , Enzimas Imobilizadas , Lipase , Lipoxigenase , Óleo de Soja , Enzimas Imobilizadas/química , Enzimas Imobilizadas/metabolismo , Lipase/química , Lipase/metabolismo , Óleo de Soja/química , Cisteína/química , Lipoxigenase/química , Lipoxigenase/metabolismo , Biocatálise , Ácidos Linoleicos/química , Peróxidos LipídicosRESUMO
The human soluble epoxide hydrolase (hsEH) is a key regulator of epoxy fatty acid (EpFA) metabolism. Inhibition of sEH can maintain endogenous levels of beneficial EpFAs and reduce the levels of their corresponding diol products, thus ameliorating a variety of pathological conditions including cardiovascular, central nervous system and metabolic diseases. The quest for orthosteric drugs that bind directly to the catalytic crevice of hsEH has been prolonged and sustained over the past decades, but the disappointing outcome of clinical trials to date warrants alternative pharmacological approaches. Previously, we have shown that hsEH can be allosterically inhibited by the endogenous electrophilic lipid 15-deoxy-Δ12,14-Prostaglandin-J2, via covalent adduction to two cysteines, C423 and C522. In this study, we explore the properties and behaviour of three electrophilic lipids belonging to the class of the nitro fatty acids, namely 9- and 10-nitrooleate and 10-nitrolinoleate. Biochemical and biophysical investigations revealed that, in addition to C423 and C522, nitro fatty acids can covalently bind to additional nucleophilic residues in hsEH C-terminal domain (CTD), two of which predicted in this study to be latent allosteric sites. Systematic mapping of the protein mutational space and evaluation of possible propagation pathways delineated selected residues, both in the allosteric patches and in other regions of the enzyme, envisaged to play a role in allosteric signalling. The responses elicited by the ligands on the covalent adduction sites supports future fragment-based design studies of new allosteric effectors for hsEH with increased efficacy and selectivity.
Assuntos
Epóxido Hidrolases , Ácidos Linoleicos , Nitrocompostos , Regulação Alostérica/efeitos dos fármacos , Cisteína/metabolismo , Epóxido Hidrolases/antagonistas & inibidores , Epóxido Hidrolases/química , Epóxido Hidrolases/metabolismo , Humanos , Ácidos Linoleicos/química , Ácidos Linoleicos/farmacologia , Nitrocompostos/química , Nitrocompostos/farmacologiaRESUMO
We describe a concise and reliable protocol for the precisely controlled tetradeuteration of straight-chain fatty acids (FAs) at the α- and ß-positions that is generally applicable to a variety of FAs, including trans-FAs, polyunsaturated FAs (PUFAs), and their oxidized derivatives. The precisely controlled introduction of four deuterium atoms into the FAs enables their persistent and quantitative tracking by LC-MS/MS analysis based on their molecular structures. In addition, the phosphatidylcholine (PC) species prepared from the tetradeuterated FAs thus obtained give a diagnostic peak, namely, a phosphocholine fragment that contains deuterium, in the LC-MS/MS analysis. With these features, the metabolism of a representative oxidized linoleic acid, that is, hydroxyoctadecadienoic acid (HODE), was investigated, leading to the identification of acyltransferases that transfer the acyl moiety derived from HODE to lysophosphatidylcholine.
Assuntos
Ácidos Graxos , Ácido Linoleico , Cromatografia Líquida , Deutério , Ácidos Linoleicos/química , Espectrometria de Massas em TandemRESUMO
Rigid polyurethane/polyisocyanurate (RPU/PIR) foam formulations were modified by evening primrose (Oenothera biennis) oil cake as a bio-filler in the amount of 5 to 50 wt.%. The obtained foams were tested in terms of processing parameters, cellular structure (SEM analysis), physico-mechanical properties (apparent density, compressive strength, brittleness, accelerated aging tests), thermal insulation properties (thermal conductivity coefficient, closed cells content, absorbability and water absorption), flammability, smoke emission, and thermal properties. The obtained results showed that the amount of bio-filler had a significant influence on the morphology of the modified foams. Thorough mixing of the polyurethane premix allowed better homogenization of the bio-filler in the polyurethane matrix, resulting in a regular cellular structure. This resulted in an improvement in the physico-mechanical and thermal insulation properties as well as a reduction in the flammability of the obtained materials. This research provided important information on the management of the waste product from the edible oil industry and the production process of fire-safe RPU/PIR foams with improved performance properties. Due to these beneficial effects, it was found that the use of evening primrose oil cake as a bio-filler for RPU/PIR foams opens a new way of waste management to obtain new "green" materials.
Assuntos
Ácidos Linoleicos/química , Oenothera biennis/química , Óleos de Plantas/química , Polímeros/química , Poliuretanos/química , Triazinas/química , Resíduos/análise , Ácido gama-Linolênico/químicaRESUMO
The widespread presence of lipid hydroperoxides in foodstuffs and biological samples has aroused great attentions in recent years, while it remains challenging for analysis of the fragility of O - O bond linkage of peroxides. In this present study, we explored the utility of electrospray ionization mass spectrometry (ESI-MS) for characterization of two fatty acid hydroperoxides from oxidation of linoleic acid and α-linolenic acid, which are the essential fatty acids abundant in many seeds and vegetable oils. The results indicated that in-source fragmentation occurred in the detection of the two fatty acid hydroperoxides in both positive and negative ion modes, which yielded characteristic fragments for ESI-MS analysis. In addition, the genotoxicity of fatty acid hydroperoxides for generation of nucleoside adducts was investigated. It was found that a variety of nucleoside adducts were formed from the reactions of fatty acid hydroperoxides and nucleosides. Furthermore, the decomposition products of the fatty acid hydroperoxides were determined, which provided evidence to elucidate the reaction mechanism for formation of nucleoside adducts.
Assuntos
Ácidos Graxos/química , Ácidos Linoleicos/química , Ácidos Linolênicos/química , Peróxidos Lipídicos/química , Nucleosídeos/química , Cromatografia Líquida de Alta Pressão/métodos , Oxirredução , Óleos de Plantas/química , Espectrometria de Massas por Ionização por Electrospray/métodosRESUMO
Lipid carriers of hydrophobic paclitaxel (PTX) are used in clinical trials for cancer chemotherapy. Improving their loading capacity requires enhanced PTX solubilization. We compared the time-dependence of PTX membrane solubility as a function of PTX content in cationic liposomes (CLs) with lipid tails containing one (oleoyl; DOPC/DOTAP) or two (linoleoyl; DLinPC/newly synthesized DLinTAP) cis double bonds by using microscopy to generate kinetic phase diagrams. The DLin lipids displayed significantly increased PTX membrane solubility over DO lipids. Remarkably, 8 mol% PTX in DLinTAP/DLinPC CLs remained soluble for approximately as long as 3 mol% PTX (the solubility limit, which has been the focus of most previous studies and clinical trials) in DOTAP/DOPC CLs. The increase in solubility is likely caused by enhanced molecular affinity between lipid tails and PTX, rather than by the transition in membrane structure from bilayers to inverse cylindrical micelles observed with small-angle X-ray scattering. Importantly, the efficacy of PTX-loaded CLs against prostate cancer cells (their IC50 of PTX cytotoxicity) was unaffected by changing the lipid tails, and toxicity of the CL carrier was negligible. Moreover, efficacy was approximately doubled against melanoma cells for PTX-loaded DLinTAP/DLinPC over DOTAP/DOPC CLs. Our findings demonstrate the potential of chemical modifications of the lipid tails to increase the PTX membrane loading while maintaining (and in some cases even increasing) the efficacy of CLs. The increased PTX solubility will aid the development of liposomal PTX carriers that require significantly less lipid to deliver a given amount of PTX, reducing side effects and costs.
Assuntos
Antineoplásicos/administração & dosagem , Ácidos Linoleicos/química , Lipossomos/química , Ácido Oleico/química , Paclitaxel/administração & dosagem , Antineoplásicos/toxicidade , Ácidos Graxos Monoinsaturados/química , Humanos , Células PC-3 , Paclitaxel/toxicidade , Fosfatidilcolinas/química , Compostos de Amônio Quaternário/químicaRESUMO
Lepidium meyenii (maca), a plant indigenous to the Peruvian Andes, recently has been utilized globally for claimed health or recreational benefits. The search for natural products that inhibit soluble epoxide hydrolase (sEH), with therapeutically relevant potencies and concentrations, led to the present study on bioactive amide secondary metabolites found in L. meyenii, the macamides. Based on known and suspected macamides, 19 possible macamides were synthesized and characterized. The majority of these amides displayed excellent inhibitory potency (IC50 ≈ 20-300 nM) toward the recombinant mouse, rat, and human sEH. Quantitative analysis of commercial maca products revealed that certain products contain known macamides (1-5, 8-12) at therapeutically relevant total concentrations (≥3.29 mg/g of root), while the inhibitory potency of L. meyenii extracts directly correlates with the sum of concentration/IC50 ratios of macamides present. Considering both its in vitro efficacy and high abundance in commercial products, N-benzyl-linoleamide (4) was identified as a particularly relevant macamide that can be utilized for in vivo studies. Following oral administration in the rat, compound 4 not only displayed acceptable pharmacokinetic characteristics but effectively reduced lipopolysaccharide-induced inflammatory pain. Inhibition of sEH by macamides provides a plausible biological mechanism of action to account for several beneficial effects previously observed with L. meyenii treatments.
Assuntos
Inibidores Enzimáticos/farmacologia , Epóxido Hidrolases/antagonistas & inibidores , Inflamação/complicações , Ácidos Linoleicos/química , Dor/prevenção & controle , Administração Oral , Analgesia , Animais , Humanos , Ácidos Linoleicos/administração & dosagem , Ácidos Linoleicos/farmacocinética , Ácidos Linoleicos/farmacologia , Camundongos , Dor/etiologia , RatosRESUMO
BACKGROUND: Studies have shown that evening primrose oil (EPO) supplementation might be effective in improving lipid profile, however, the results are inconsistent. This study was performed to determine the direction and magnitude of the EPO effect on the lipid profile. METHODS: PubMed, Scopus, Cochrane Library, Embase and Web of Science databases and Google Scholar were searched up to September-2019. Meta-analysis was performed using the random-effects model. Lipid profile including high-density lipoprotein (HDL), total cholesterol (TC), triglyceride (TG), and low-density lipoprotein (LDL) was considered as the primary outcome. RESULTS: A total of 926 articles were identified through database searching, of which, six RCTs were included in the meta-analysis. There were six studies on HDL, TC, and TG and four studies on LDL. EPO supplementation had no significant effect on TC, TG, LDL, and HDL. However, in subgroup analysis, a significant reduction in TG at a dose of ≤4 g/day (weighted mean difference [WMD] = -37.28 mg/dl; 95% CI: -73.53 to -1.03, p = .044) and a significant increase in HDL in hyperlipidemic subjects (WMD = 5.468 mg/dl; 95% CI: 1.323 to 9.614, p = .010) was found. CONCLUSION: Oral intake of EPO at a dose of ≤4 g/day significantly reduces serum TG levels and significantly increases HDL levels in hyperlipidemic subjects.
Assuntos
Ácidos Linoleicos/química , Metabolismo dos Lipídeos/efeitos dos fármacos , Lipídeos/química , Óleos de Plantas/química , Ácido gama-Linolênico/química , Humanos , Oenothera biennis , Ensaios Clínicos Controlados Aleatórios como AssuntoRESUMO
Hydroxyoctadecadienoic acids (HODEs) are produced by oxidation and reduction of linoleates. There are several regio- and stereo-isomers of HODE, and their concentrations in vivo are higher than those of other lipids. Although conformational isomers may have different biological activities, comparative analysis of intracellular function of HODE isomers has not yet been performed. We evaluated the transcriptional activity of peroxisome proliferator-activated receptor γ (PPARγ), a therapeutic target for diabetes, and analyzed PPARγ agonist activity of HODE isomers. The lowest scores for docking poses of 12 types of HODE isomers (9-, 10-, 12-, and 13-HODEs) were almost similar in docking simulation of HODEs into PPARγ ligand-binding domain (LBD). Direct binding of HODE isomers to PPARγ LBD was determined by water-ligand observed via gradient spectroscopy (WaterLOGSY) NMR experiments. In contrast, there were differences in PPARγ agonist activities among 9- and 13-HODE stereo-isomers and 12- and 13-HODE enantio-isomers in a dual-luciferase reporter assay. Interestingly, the activity of 9-HODEs was less than that of other regio-isomers, and 9-(E,E)-HODE tended to decrease PPARγ-target gene expression during the maturation of 3T3-L1 cells. In addition, 10- and 12-(Z,E)-HODEs, which we previously proposed as biomarkers for early-stage diabetes, exerted PPARγ agonist activity. These results indicate that all HODE isomers have PPARγ-binding affinity; however, they have different PPARγ agonist activity. Our findings may help to understand the biological function of lipid peroxidation products.
Assuntos
Ácidos Linoleicos/farmacologia , PPAR gama/agonistas , Células 3T3-L1 , Animais , Ácidos Linoleicos/química , Peroxidação de Lipídeos , Camundongos , Simulação de Acoplamento Molecular , Estrutura Molecular , PPAR gama/química , PPAR gama/metabolismo , Estereoisomerismo , Relação Estrutura-AtividadeRESUMO
SCOPE: The docosahexaenoic acid ester of hydroxy linoleic acid (13-DHAHLA) is a bioactive lipid with anti-inflammatory properties from the family of fatty acid esters of hydroxy fatty acids (FAHFA). METHODS AND RESULTS: To explore the biosynthesis of 13-DHAHLA from dietary oils, C57BL/6N mice are gavaged for 8 days with various corn oil/marine oil mixtures containing the same amount of DHA. Plasma levels of omega-3 FAHFAs are influenced by the lipid composition of the mixtures but do not reflect the changes in bioavailability of polyunsaturated fatty acids in plasma. Triacylglycerol-bound DHA and linoleic acid serve as more effective precursors for 13-DHAHLA synthesis than DHA bound in phospholipids or wax esters. Both 13(S)- and 13(R)-DHAHLA inhibit antigen and PGE2 -induced chemotaxis and degranulation of mast cells to a comparable extent and 13(S)-DHAHLA is identified as the predominant isomer in mouse adipose tissue. CONCLUSION: Here, the optimal nutritional source of DHA is identified, which supports production of anti-inflammatory FAHFAs, as triacylglycerol-based marine oil and also reveals a possible role of triacylglycerols in the synthesis of FAHFA lipokines.
Assuntos
Anti-Inflamatórios não Esteroides/farmacocinética , Ácidos Docosa-Hexaenoicos/farmacocinética , Óleos/química , Óleos/farmacocinética , Animais , Anti-Inflamatórios não Esteroides/sangue , Organismos Aquáticos , Disponibilidade Biológica , Quimiotaxia/efeitos dos fármacos , Ácidos Graxos Ômega-3/farmacocinética , Ácidos Graxos Ômega-3/farmacologia , Feminino , Ácidos Linoleicos/química , Masculino , Mastócitos/efeitos dos fármacos , Camundongos Endogâmicos C57BL , Estereoisomerismo , Triglicerídeos/químicaRESUMO
The induction period (IP) of ethyl linoleate stressed at 60 °C was monitored via the formation of hydroperoxides. The addition of lycopene (1% w/w) increased the IP from 7.0 to 10.0 h to prove the strong antioxidative potential in contrast to ß-carotene with pro-oxidative effects (IP: 6.0 h), both showing strong scavenging activity under fast degradation. When peroxidation was induced by singlet oxygen, both carotenoids effectively inhibited the formation of hydroperoxides, with quenching activity only observed at low singlet oxygen concentrations, while scavenging still dominated. Thus, carotenoids did not interact with the introduced singlet oxygen but rather with the radical intermediates of fat oxidation. These experiments were then transferred to lecithin-based micelles more related to biological systems, where singlet oxygen was generated in the outer aqueous phase. Lycopene and ß-carotene delayed or inhibited lipid peroxidation depending on concentration. In this setup, ß-carotene showed exclusively quenching activity, while lycopene was additionally degraded to about 70%.
Assuntos
Ácidos Linoleicos/química , Licopeno/química , beta Caroteno/química , Antioxidantes/química , Antioxidantes/farmacologia , Cinética , Lecitinas/química , Ácidos Linoleicos/farmacologia , Peroxidação de Lipídeos/efeitos dos fármacos , Modelos Químicos , Oxirredução , Oxigênio Singlete/químicaRESUMO
Nonalcoholic steatohepatitis (NASH) is a common liver disease that occurs in both alcoholics and nonalcoholics. Oxidative stress is a possible causative factor for liver diseases including NASH. Gut microorganisms, especially lactic acid bacteria, can produce unique fatty acids, including hydroxy, oxo, conjugated, and partially saturated fatty acids. The oxo fatty acid 10-oxo-11(E)-octadecenoic acid (KetoC) provides potent cytoprotective effects against oxidative stress through activation of Nrf2-ARE pathway. The aim of this study was to explore the preventive and therapeutic effects of gut microbial fatty acid metabolites in a NASH mouse model. The mice were divided into 3 experimental groups and fed as follows: (1) high-fat diet (HFD) (2) HFD mixed with 0.1% KetoA (10-oxo-12(Z)-octadecenoic acid), and (3) HFD mixed with 0.1% KetoC. After 3 weeks of feeding, plasma parameters, liver histology, and mRNA expression of multiple genes were assessed. There was hardly any difference in fat accumulation in the histological study; however, no ballooning occurred in 2/5 mice of KetoC group. Bridging fibrosis was not observed in the KetoA group, although KetoA administration did not significantly suppress fibrosis score (p = 0.10). In addition, KetoC increased the expression level of HDL related genes and HDL cholesterol levels in the plasma. These results indicated that KetoA and KetoC may partly affect the progression of NASH in mice models.
Assuntos
Bactérias/metabolismo , Redes Reguladoras de Genes/efeitos dos fármacos , Ácidos Linoleicos/administração & dosagem , Hepatopatia Gordurosa não Alcoólica/dietoterapia , Animais , HDL-Colesterol/sangue , Dieta Hiperlipídica/efeitos adversos , Modelos Animais de Doenças , Progressão da Doença , Microbioma Gastrointestinal , Perfilação da Expressão Gênica , Regulação da Expressão Gênica/efeitos dos fármacos , Células Hep G2 , Humanos , Ácidos Linoleicos/sangue , Ácidos Linoleicos/química , Ácidos Linoleicos/farmacologia , Masculino , Camundongos , Hepatopatia Gordurosa não Alcoólica/induzido quimicamente , Hepatopatia Gordurosa não Alcoólica/genética , Estresse Oxidativo/efeitos dos fármacos , Estreptozocina/efeitos adversosRESUMO
In this study, the effects of limited and excess phosphate on biomass content, oil content, fatty acid profile and the expression of three fatty acid desaturases in Messastrum gracile SE-MC4 were determined. It was found that total biomass (0.67-0.83 g L-1), oil content (30.99-38.08%) and the duration for cells to reach stationary phase (25-27 days) were not considerably affected by phosphate limitation. However, excess phosphate slightly reduced total biomass and oil content to 0.50 g L-1 and 25.36% respectively. The dominant fatty acids in M. gracile, pamitic acid (C16:0) and oleic acid (C18:1) which constitute more than 81% of the total fatty acids remained relatively high and constant across all phosphate concentrations. Reduction of phosphate concentration to 25% and below significantly increased total MUFA, whereas increasing phosphate concentration to ≥ 50% and ≥ 100% significantly increased total SFA and PUFA content respectively. The expression of omega-3 fatty acid desaturase (ω-3 FADi1, ω-3 FADi2) and omega-6 fatty acid desaturase (ω-6 FAD) was increased under phosphate limitation, especially at ≤ 12.5% phosphate, whereas levels of streoyl-ACP desaturase (SAD) transcripts were relatively unchanged across all phosphate concentrations. The first isoform of ω-3 FAD (ω-3 FADi) displayed a binary upregulation under limited (≤ 12.5%) and excess (200%) phosphate. The expression of ω-6 FAD, ω-3 FAD and SAD were inconsistent with the accumulation of oleic acid (C18:1), linoleic acid (C18:2) and alpha-linolenic acid (C18:3), suggesting that these genes may be regulated indirectly by phosphate availability via post-transcriptional or post-translational mechanisms.
Assuntos
Clorofíceas/enzimologia , Ácidos Graxos Dessaturases/metabolismo , Ácidos Graxos/química , Ácidos Linoleicos/química , Lipídeos/química , Fosfatos/química , Biomassa , Ácidos Graxos Dessaturases/genética , Regulação Enzimológica da Expressão Gênica , Microbiologia Industrial/métodos , Óleos , Isoformas de ProteínasRESUMO
PURPOSE: We hypothesize that different types of dietary fatty acids (FAs) affect gastrointestinal (GI) motility and visceromotor function and that this effect can be regulated by the fatty acid binding protein 4 (FABP4). METHODS: Mice were fed for 60 days with standard diet (STD), STD with 7% (by weight) coconut oil, rich in medium-chain FAs (MCFAs) (COCO), or with 7% evening primrose oil, rich in long-chain FAs (LCFAs) (EPO). In each group, half of the mice received FABP4 inhibitor, BMS309403 (1 mg/kg; i.p.) twice a week. Body weight (BW) and food intake were measured; well-established tests were performed to characterize the changes in GI motility and visceral pain. White adipose tissue and colonic samples were collected for cell culturing and molecular studies. RESULTS: COCO significantly increased GI transit, but not colonic motility. COCO and EPO delayed the onset of diarrhea, but none affected the effect of loperamide. EPO reduced BW and increased the visceromotor response (VMR) to colorectal distension (CRD). COCO and EPO reduced differentiation of preadipocytes. Treatment with BMS309403: (1) reversed the effects induced by COCO in physiological conditions and in mouse models of diarrhea; (2) prevented the effects of EPO on BW, VMR to CRD and castor oil-induced diarrhea; (3) affected proliferation of preadipocytes; (4) changed the expression of Fabp4 in colonic and adipocyte samples from COCO and EPO. CONCLUSION: Modifying dietary intake of MCFAs and LCFAs may be used to control GI motility or visceral pain and thus modulate the symptoms of functional GI disorders. The effect is dependent on the expression of FABP4.
Assuntos
Gorduras na Dieta/farmacologia , Proteínas de Ligação a Ácido Graxo/metabolismo , Ácidos Graxos/química , Ácidos Graxos/farmacologia , Motilidade Gastrointestinal/efeitos dos fármacos , Dor Visceral/dietoterapia , Animais , Óleo de Coco/química , Óleo de Coco/farmacologia , Diarreia/dietoterapia , Dietoterapia , Proteínas de Ligação a Ácido Graxo/antagonistas & inibidores , Trânsito Gastrointestinal/efeitos dos fármacos , Ácidos Linoleicos/química , Ácidos Linoleicos/farmacologia , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Oenothera biennis , Óleos de Plantas/química , Óleos de Plantas/farmacologia , Ácido gama-Linolênico/química , Ácido gama-Linolênico/farmacologiaRESUMO
Anandamide (AEA) played potent neuroprotective activities via cannabinoid type 1 (CB1) and 2 (CB2) receptor. N-Linoleyltyrosine (NITyr), as an AEA analogue, was synthesized in our laboratory and evaluated the neuroprotective effects and mechanisms for the first time. NITyr was synthesized via substitution reaction. The neuroprotective effects of NITyr were evaluated in a gerbil model of transient cerebral ischemia. Each gerbil was subjected to open field test (OFT), Rotard rod test (RRT), Morris water maze (MWM) successively and executed after animal behaviors. Part of the brain was stained with hematoxylin and eosin (HE) and Nissl staining, and the rest for biochemical analysis. NITyr could not increase spontaneous locomotor activity and ameliorate the anxiety behavior in the OFT but could improve the motor coordination in the RRT and the spatial memory impairment in the MWM. Immunohistochemically, NITyr could attenuate the ischemia-induced neural loss in the hippocampus. The Enzyme-linked immunosorbent assay (ELISA) suggested that NITyr ameliorated the inflammation and oxidative stress. Consistently, NITyr could up-regulate the expressions of p-phosphadylinositol 3-kinase (PI3K) and p-Akt but not PI3K and Akt in the hippocampus. In addition to oxidative stress, CB2 receptor antagonist AM630 but not CB1 receptor antagonist AM251 could reverse the above phenomena. However, CB1 receptor antagonist AM251 could reverse oxidative stress. Accordingly, NITyr could up-regulate the expressions of CB2 but not CB1. NITyr could improve the motor coordination, learning and memory impairments, neural loss in the hippocampus and the inflammation of the mice via CB2 receptor involvement of PI3K/Akt signaling pathway.
Assuntos
Ataque Isquêmico Transitório/tratamento farmacológico , Ácidos Linoleicos/farmacologia , Fosfatidilinositol 3-Quinases/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Receptor CB2 de Canabinoide/metabolismo , Tirosina/análogos & derivados , Animais , Ansiedade/tratamento farmacológico , Gerbillinae , Hipocampo/patologia , Ácidos Linoleicos/química , Masculino , Aprendizagem em Labirinto , Atividade Motora/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Receptor CB1 de Canabinoide/metabolismo , Teste de Desempenho do Rota-Rod , Transdução de Sinais/efeitos dos fármacos , Aprendizagem Espacial/efeitos dos fármacos , Tirosina/química , Tirosina/farmacologiaRESUMO
Plant lipoxygenases oxygenate linoleic acid to produce 13(S)-hydroperoxy-9Z,11E-octadecadienoic acid (13(S)-HPOD) or 9-hydroperoxy-10E,12Z-octadecadienoic acid (9(S)-HPOD). The manner in which these enzymes bind substrates and the mechanisms by which they control regiospecificity are uncertain. Hornung et al. (Proc. Natl. Acad. Sci. USA96 (1999) 4192-4197) have identified an important residue, corresponding to phe-557 in soybean lipoxygenase-1 (SBLO-1). These authors proposed that large residues in this position favored binding of linoleate with the carboxylate group near the surface of the enzyme (tail-first binding), resulting in formation of 13(S)-HPOD. They also proposed that smaller residues in this position facilitate binding of linoleate in a head-first manner with its carboxylate group interacting with a conserved arginine residue (arg-707 in SBLO-1), which leads to 9(S)-HPOD. In the present work, we have tested these proposals on SBLO-1. The F557V mutant produced 33% 9-HPOD (S:Râ¯=â¯87:13) from linoleic acid at pH 7.5, compared with 8% for the wild-type enzyme and 12% with the F557V,R707L double mutant. Experiments with 11(S)-deuteriolinoleic acid indicated that the 9(S)-HPOD produced by the F557V mutant involves removal of hydrogen from the pro-R position on C-11 of linoleic acid, as expected if 9(S)-HPOD results from binding in an orientation that is inverted relative to that leading to 13(S)-HPOD. The product distributions obtained by oxygenation of 10Z,13Z-nonadecadienoic acid and arachidonic acid by the F557V mutant support the hypothesis that ω6 oxygenation results from tail-first binding and ω10 oxygenation from head-first binding. The results demonstrate that the regiospecificity of SBLO-1 can be altered by a mutation that facilitates an alternative mode of substrate binding and adds to the body of evidence that 13(S)-HPOD arises from tail-first binding.
Assuntos
Ácidos Graxos Insaturados/metabolismo , Glycine max/enzimologia , Lipoxigenase/metabolismo , Sítios de Ligação , Catálise , Deutério/química , Ácidos Graxos Insaturados/química , Ácidos Linoleicos/química , Peróxidos Lipídicos/química , Lipoxigenase/genética , Mutação , Oxirredução , Fosfatidilcolinas/química , Ligação Proteica , EstereoisomerismoRESUMO
This paper proposes the use of random forest for adulteration detection purposes, combining the random forest algorithm with the artificial generation of outliers from the authentic samples. This proposal was applied in two food adulteration studies: evening primrose oils using ATR-FTIR spectroscopy and ground nutmeg using NIR diffuse reflectance spectroscopy. The primrose oil was adulterated with soybean, corn and sunflower oils, and the model was validated using these adulterated oils and other different oils, such as rosehip and andiroba, in pure and adulterated forms. The ground nutmeg was adulterated with cumin, commercial monosodium glutamate, soil, roasted coffee husks and wood sawdust. For the primrose oil, the proposed method presented superior performance than PLS-DA and similar performance to SIMCA and for the ground nutmeg, the random forest was superior to PLS-DA and SIMCA. Also, in both applications using the random forest, no sample was excluded from the external validation set.
Assuntos
Contaminação de Alimentos/análise , Ácidos Linoleicos/química , Óleos de Plantas/química , Espectroscopia de Infravermelho com Transformada de Fourier/métodos , Ácido gama-Linolênico/química , Óleo de Milho/análise , Limite de Detecção , Myristica/química , Oenothera biennis , Óleo de Soja/análise , Óleo de Girassol/análiseRESUMO
Fatty acid esters of hydroxy fatty acids (FAHFAs) are a recently discovered class of biologically active lipids. Here we identify the linoleic acid ester of 13-hydroxy linoleic acid (13-LAHLA) as an anti-inflammatory lipid. An oat oil fraction and FAHFA-enriched extract from this fraction showed anti-inflammatory activity in a lipopolysaccharide-induced cytokine secretion assay. Structural studies identified three LAHLA isomers (15-, 13-, and 9-LAHLA) as being the most abundant FAHFAs in the oat oil fraction. Of these LAHLAs, 13-LAHLA is the most abundant LAHLA isomer in human serum after ingestion of liposomes made of fractionated oat oil, and it is also the most abundant endogenous LAHLA in mouse and human adipose tissue. As a result, we chemically synthesized 13-LAHLA for biological assays. 13-LAHLA suppresses lipopolysaccharide-stimulated secretion of cytokines and expression of pro-inflammatory genes. These studies identify LAHLAs as an evolutionarily conserved lipid with anti-inflammatory activity in mammalian cells.
Assuntos
Anti-Inflamatórios/química , Avena/química , Ésteres/química , Ácidos Linoleicos/química , Tecido Adiposo/química , Tecido Adiposo/metabolismo , Animais , Anti-Inflamatórios/análise , Anti-Inflamatórios/farmacologia , Avena/metabolismo , Proliferação de Células/efeitos dos fármacos , Cromatografia Líquida de Alta Pressão , Citocinas/metabolismo , Humanos , Lipopolissacarídeos/toxicidade , Macrófagos/citologia , Macrófagos/efeitos dos fármacos , Macrófagos/metabolismo , Espectrometria de Massas , Camundongos , Óleos de Plantas/química , Óleos de Plantas/farmacologia , Células RAW 264.7 , EstereoisomerismoRESUMO
A Chinese-style sausage was processed using pork as the raw material. During the whole process, 13-hydroxyoctadecadienoic acid (13-HODE), 9-hydroxyoctadecadienoic acid (9-HODE), 9,10-dihydroxyoctadecenoic acid (9,10-DHODE) and 9,10,13-trihydroxyoctadecenoic acid (9,10,13-THODE) kept increasing. All of them were found to be correlated negatively and significantly with lipoxygenases (LOX) activity, and positively and significantly with peroxide value (POV) and thiobarbituric acid reactive substances (TBARS). The ratio of 13-HODE to 9-HODE decreased slowly during drying stage and stayed higher than 2 during the whole process, and it was found to be positively and significantly with LOX activity. The ratio of variation of 13-HODE to variation of 9-HODE in every sampling period (the ratio of Δ13-HODE to Δ9-HODE) decreased sharply from 2.75 in the stage of curing for 12â¯h to 1.37 in the stage drying from 24 d to 30 d. The changes of ratio of 13HODE to 9-HODE and ratio of Δ13-HODE to Δ9-HODE indicated LOX-catalyzed oxidation predominated in curing and early drying stages, and such predominance was taken over by non-enzymatic oxidation during late drying stage; LOX-catalyzed oxidation was the major contributor to lipids oxidation during the whole process of the Chinese-style sausage preparing.