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1.
Nutrients ; 11(9)2019 Sep 11.
Artigo em Inglês | MEDLINE | ID: mdl-31514292

RESUMO

The use of a complete nutritional approach seems increasingly promising to combat chronic inflammation. The choice of healthy sources of carbohydrates, fats, and proteins, associated with regular physical activity and avoidance of smoking is essential to fight the war against chronic diseases. At the base of the analgesic, anti-inflammatory, or antioxidant action of the diets, there are numerous molecules, among which some of a lipidic nature very active in the inflammatory pathway. One class of molecules found in diets with anti-inflammatory actions are ALIAmides. Among all, one is particularly known for its ability to counteract the inflammatory cascade, the Palmitoylethanolamide (PEA). PEA is a molecular that is present in nature, in numerous foods, and is endogenously produced by our body, which acts as a balancer of inflammatory processes, also known as endocannabionoid-like. PEA is often used in the treatment of both acute and chronic inflammatory pathologies, either alone or in association with other molecules with properties, such as antioxidants or analgesics. This review aims to illustrate an overview of the different diets that are involved in the process of opposition to the inflammatory cascade, focusing on capacity of PEA and new formulations in synergy with other molecules.


Assuntos
Anti-Inflamatórios/uso terapêutico , Antioxidantes/uso terapêutico , Suplementos Nutricionais , Etanolaminas/uso terapêutico , Inflamação/prevenção & controle , Ácidos Palmíticos/uso terapêutico , Animais , Anti-Inflamatórios/efeitos adversos , Antioxidantes/efeitos adversos , Suplementos Nutricionais/efeitos adversos , Sinergismo Farmacológico , Etanolaminas/efeitos adversos , Etanolaminas/metabolismo , Humanos , Inflamação/metabolismo , Mediadores da Inflamação/metabolismo , Ácidos Palmíticos/efeitos adversos , Ácidos Palmíticos/metabolismo , Transdução de Sinais
2.
BMC Cancer ; 19(1): 823, 2019 Aug 20.
Artigo em Inglês | MEDLINE | ID: mdl-31429730

RESUMO

BACKGROUND: Natural killer (NK) cell dysfunction following cancer surgery has been shown to promote metastases. Recent studies demonstrate an emerging role for lipids in the modulation of NK cell innate responses. However, the mechanisms involved in lipid modulation of NK cell postoperative anti-tumor function are unknown. This current study will determine whether the lipid accumulation via scavenger receptors on NK cells is responsible for the increase in postoperative metastasis. METHODS: Lipid content in mouse and human NK cells was evaluated by flow cytometry. NK cell scavenger receptor (SR) expression was measured by microarray analysis, validated by qRT-PCR and flow cytometry. NK cell ex vivo and in vivo tumor killing was measured by chromium-release and adoptive transfer assays, respectively. The mediating role of surgery-expanded granulocytic myeloid derived suppressor cells (gMDSC) in SR induction on NK cells was evaluated using co-culture assays. RESULTS: NK cells in surgery-treated mice demonstrated increased lipid accumulation, which occurred via up-regulation of MSR1, CD36 and CD68. NK cells with high lipid content had diminished ability to lyse tumor targets ex vivo. Adoptive transfer of lipid-laden NK cells into NK cell-deficient mice were unable to protect against a lung tumor challenge. Granulocytic MDSC from surgery-treated mice increased SR expression on NK cells. Colorectal cancer surgical patients showed increased NK cell lipid content, higher CD36 expression, decreased granzyme B and perforin production in addition to reduced cytotoxicity in the postoperative period. CONCLUSIONS: Postoperative lipid accumulation promotes the formation of metastases by impairing NK cell function in both preclinical surgical models and human surgical colorectal cancer patient samples. Understanding and targeting the mechanisms underlying lipid accumulation in innate immune NK cells can improve prognosis in cancer surgical patients.


Assuntos
Neoplasias Colorretais/metabolismo , Citotoxicidade Imunológica , Células Matadoras Naturais/imunologia , Células Matadoras Naturais/metabolismo , Ácidos Palmíticos/metabolismo , Transferência Adotiva , Animais , Antígenos CD/genética , Antígenos de Diferenciação Mielomonocítica/genética , Antígenos CD36/genética , Neoplasias Colorretais/cirurgia , Modelos Animais de Doenças , Feminino , Granzimas/metabolismo , Humanos , Células K562 , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Perforina/metabolismo , Período Pós-Operatório , Receptores Depuradores/genética , Receptores Depuradores Classe A/genética
3.
Medicina (Kaunas) ; 55(4)2019 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-30939862

RESUMO

Asthma is a common allergic pathology of the respiratory tract that requires the study of mechanisms underlying it, due to severe forms of the disease, which are refractory to therapy. The review is devoted to the search for molecular targets of fatty acid ethanolamides in asthma, in particular palmitoylethanolamide (PEA), which has been successfully used in the treatment of chronic inflammatory and neurodegenerative diseases, in the pathogenesis of which the nervous and immune systems are involved. Recently, the potentially important role of neuro-immune interactions in the development of allergic reactions has been established. Many of the clinical symptoms accompanying allergic airway inflammation are the result of the activation of neurons in the airways, so the attention of researchers is currently focused on neuro-immune interactions, which can play an important role in asthma pathophysiology. A growing number of scientific works confirm that the key molecule in the implementation of these inter-systemic interactions is nerve growth factor (NGF). In addition to its classic role in nervous system physiology, NGF is considered as an important factor associated with the pathogenesis of allergic diseases, particularly asthma, by regulating of mast cell differentiation. In this regard, NGF can be one of the targets of PEA in asthma therapy. PEA has a biological effect on the nervous system, and affects the activation and the degranulation of mast cells.


Assuntos
Anti-Inflamatórios não Esteroides/uso terapêutico , Asma/tratamento farmacológico , Asma/metabolismo , Etanolaminas/uso terapêutico , Terapia de Alvo Molecular , Fator de Crescimento Neural/metabolismo , Ácidos Palmíticos/uso terapêutico , Animais , Anti-Inflamatórios não Esteroides/metabolismo , Asma/imunologia , Eosinófilos/metabolismo , Etanolaminas/metabolismo , Ácidos Graxos/metabolismo , Humanos , Hipersensibilidade/metabolismo , Inflamação/imunologia , Inflamação/metabolismo , Mastócitos/metabolismo , Camundongos , Neurônios/metabolismo , Ácidos Palmíticos/metabolismo
4.
Nature ; 566(7744): 403-406, 2019 02.
Artigo em Inglês | MEDLINE | ID: mdl-30728499

RESUMO

Most tumours have an aberrantly activated lipid metabolism1,2 that enables them to synthesize, elongate and desaturate fatty acids to support proliferation. However, only particular subsets of cancer cells are sensitive to approaches that target fatty acid metabolism and, in particular, fatty acid desaturation3. This suggests that many cancer cells contain an unexplored plasticity in their fatty acid metabolism. Here we show that some cancer cells can exploit an alternative fatty acid desaturation pathway. We identify various cancer cell lines, mouse hepatocellular carcinomas, and primary human liver and lung carcinomas that desaturate palmitate to the unusual fatty acid sapienate to support membrane biosynthesis during proliferation. Accordingly, we found that sapienate biosynthesis enables cancer cells to bypass the known fatty acid desaturation pathway that is dependent on stearoyl-CoA desaturase. Thus, only by targeting both desaturation pathways is the in vitro and in vivo proliferation of cancer cells that synthesize sapienate impaired. Our discovery explains metabolic plasticity in fatty acid desaturation and constitutes an unexplored metabolic rewiring in cancers.


Assuntos
Ácidos Graxos/química , Ácidos Graxos/metabolismo , Redes e Vias Metabólicas , Neoplasias/metabolismo , Neoplasias/patologia , Animais , Linhagem Celular Tumoral , Membrana Celular/metabolismo , Proliferação de Células , Ácidos Graxos Dessaturases/metabolismo , Feminino , Células HEK293 , Humanos , Masculino , Camundongos , Ácidos Oleicos/metabolismo , Palmitatos/metabolismo , Ácidos Palmíticos/metabolismo , Estearoil-CoA Dessaturase/metabolismo
5.
Int J Mol Sci ; 20(4)2019 Feb 15.
Artigo em Inglês | MEDLINE | ID: mdl-30769921

RESUMO

Palmitic acid metabolism involves delta-9 and delta-6 desaturase enzymes forming palmitoleic acid (9cis-16:1; n-7 series) and sapienic acid (6cis-16:1; n-10 series), respectively. The corresponding biological consequences and lipidomic research on these positional monounsaturated fatty acid (MUFA) isomers are under development. Furthermore, sapienic acid can bring to the de novo synthesis of the n-10 polyunsaturated fatty acid (PUFA) sebaleic acid (5cis,8cis-18:2), but such transformations in cancer cells are not known. The model of Caco-2 cell line was used to monitor sapienic acid supplementation (150 and 300 µM) and provide evidence of the formation of n-10 fatty acids as well as their incorporation at levels of membrane phospholipids and triglycerides. Comparison with palmitoleic and palmitic acids evidenced that lipid remodelling was influenced by the type of fatty acid and positional isomer, with an increase of 8cis-18:1, n-10 PUFA and a decrease of saturated fats in case of sapienic acid. Cholesteryl esters were formed only in cases with sapienic acid. Sapienic acid was the less toxic among the tested fatty acids, showing the highest EC50s and inducing death only in 75% of cells at the highest concentration tested. Two-photon fluorescent microscopy with Laurdan as a fluorescent dye provided information on membrane fluidity, highlighting that sapienic acid increases the distribution of fluid regions, probably connected with the formation of 8cis-18:1 and the n-10 PUFA in cell lipidome. Our results bring evidence for MUFA positional isomers and de novo PUFA synthesis for developing lipidomic analysis and cancer research.


Assuntos
Neoplasias do Colo/metabolismo , Ácidos Graxos Ômega-3/metabolismo , Ácidos Palmíticos/metabolismo , Fosfolipídeos/química , Células CACO-2 , Membrana Celular/química , Membrana Celular/metabolismo , Ésteres do Colesterol/biossíntese , Ésteres do Colesterol/química , Ésteres do Colesterol/metabolismo , Neoplasias do Colo/química , Neoplasias do Colo/patologia , Ácidos Graxos Monoinsaturados/química , Ácidos Graxos Monoinsaturados/farmacologia , Ácidos Graxos Ômega-3/biossíntese , Humanos , Ácidos Linoleicos/química , Ácidos Linoleicos/metabolismo , Ácidos Linoleicos/farmacologia , Linoleoil-CoA Desaturase/química , Microscopia de Fluorescência , Ácido Palmítico/química , Ácido Palmítico/metabolismo , Ácidos Palmíticos/química , Ácidos Palmíticos/farmacologia , Fosfolipídeos/biossíntese
6.
Food Chem ; 283: 331-337, 2019 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-30722880

RESUMO

Structured TAGs with palmitic acid and polyunsaturated fatty acid (PUFA) at the sn-2 position have various health benefits for infants. In this study, we first compared two enzymatic routes for preparation of the structured TAGs. Results showed that the one-pot and two-step syntheses led to 37.6% and 55.4% oleic acid incorporation, respectively, after 10 h and reaction route had little effect on the sn-2 fatty acid composition. Subsequently, reaction variables of the two-step synthesis were optimized. Under the optimal conditions, 53.5% oleic acid was incorporated into the structured TAGs after 6-h acidolysis. Major fatty acids at the sn-2 position were palmitic acid (68.7%), ARA (9.8%) and oleic acid (7.9%). This is the first study reporting a two-step enzymatic method for structured TAGs preparation. Compared to the one-pot synthesis, current method significantly improves the efficiency of the acidolysis by product inhibition elimination. The synthetic TAGs have potential use in infant formulas.


Assuntos
Ácido Araquidônico/metabolismo , Fórmulas Infantis/análise , Ácidos Palmíticos/metabolismo , Triglicerídeos/química , Ácido Araquidônico/química , Ácidos Graxos Insaturados/química , Ácidos Graxos Insaturados/metabolismo , Humanos , Lactente , Lipase/metabolismo , Ácido Oleico/química , Ácido Oleico/metabolismo , Ácidos Palmíticos/química , Triglicerídeos/metabolismo
7.
Neuropharmacology ; 146: 212-221, 2019 03 01.
Artigo em Inglês | MEDLINE | ID: mdl-30496751

RESUMO

In agreement with the neurodevelopmental hypothesis of schizophrenia, prenatal exposure of rats to the antimitotic agent methylazoxymethanol acetate (MAM) at gestational day 17 produced long-lasting behavioral alterations such as social withdrawal and cognitive impairment in the social interaction test and in the novel object recognition test, respectively. At the molecular level, an increased cannabinoid receptor type-1 (CB1) mRNA and protein expression, which might be due to reduction in DNA methylation at the gene promoter in the prefrontal cortex (PFC), coincided with deficits in the social interaction test and in the novel object recognition test in MAM rats. Both the schizophrenia-like phenotype and altered transcriptional regulation of CB1 receptors were reversed by peripubertal treatment (from PND 19 to PND 39) with the non-psychotropic phytocannabinoid cannabidiol (30 mg/kg/day), or, in part, by treatment with the cannabinoid CB1 receptor antagonist/inverse agonist AM251 (0.5 mg/kg/day), but not with haloperidol (0.6 mg/kg/day). These results suggest that early treatment with cannabidiol may prevent both the appearance of schizophrenia-like deficits as well as CB1 alterations in the PFC at adulthood, supporting that peripubertal cannabidiol treatment might be protective against MAM insult.


Assuntos
Canabidiol/farmacologia , Acetato de Metilazoximetanol/farmacologia , Efeitos Tardios da Exposição Pré-Natal/tratamento farmacológico , Esquizofrenia/tratamento farmacológico , Animais , Ácidos Araquidônicos/metabolismo , Modelos Animais de Doenças , Endocanabinoides/metabolismo , Etanolaminas/metabolismo , Feminino , Glicerídeos/metabolismo , Hipocampo/metabolismo , Relações Interpessoais , Masculino , Atividade Motora/efeitos dos fármacos , Ácidos Oleicos/metabolismo , Ácidos Palmíticos/metabolismo , Piperidinas/farmacologia , Alcamidas Poli-Insaturadas/metabolismo , Córtex Pré-Frontal/metabolismo , Gravidez , Efeitos Tardios da Exposição Pré-Natal/metabolismo , Puberdade , Pirazóis/farmacologia , RNA Mensageiro/metabolismo , Ratos , Receptor CB1 de Canabinoide/metabolismo , Esquizofrenia/induzido quimicamente , Esquizofrenia/metabolismo
8.
Mol Cell Biol ; 39(5)2019 03 01.
Artigo em Inglês | MEDLINE | ID: mdl-30530523

RESUMO

Phytosphingosine (PHS) is the major long-chain base component of sphingolipids in Saccharomyces cerevisiae The PHS metabolic pathway includes a fatty acid (FA) α-oxidation reaction. Recently, we identified the novel protein Mpo1, which is involved in PHS metabolism. However, the details of the FA α-oxidation reaction and the role of Mpo1 in PHS metabolism remained unclear. In the present study, we revealed that Mpo1 is involved in the α-oxidation of 2-hydroxy (2-OH) palmitic acid (C16:0-COOH) in the PHS metabolic pathway. Our in vitro assay revealed that not only the Mpo1-containing membrane fraction but also the soluble fraction was required for the α-oxidation of 2-OH C16:0-COOH. The addition of Fe2+ eliminated the need for the soluble fraction. Purified Mpo1 converted 2-OH C16:0-COOH to C15:0-COOH in the presence of Fe2+, indicating that Mpo1 is the enzyme body responsible for catalyzing the FA α-oxidation reaction. This reaction was also found to require an oxygen molecule. Our findings indicate that Mpo1 catalyzes the FA α-oxidation reaction as 2-OH fatty acid dioxygenase, mediated by iron(IV) peroxide. Although numerous Mpo1 homologs exist in bacteria, fungi, protozoa, and plants, their functions had not yet been clarified. However, our findings suggest that these family members function as dioxygenases.


Assuntos
Ácidos Palmíticos/metabolismo , Esfingolipídeos/metabolismo , Esfingosina/análogos & derivados , Catálise , Dioxigenases/metabolismo , Ácidos Graxos/metabolismo , Ferro/metabolismo , Oxirredução , Saccharomyces cerevisiae/metabolismo , Proteínas de Saccharomyces cerevisiae/metabolismo , Esfingosina/metabolismo
9.
Nutrients ; 10(11)2018 Nov 13.
Artigo em Inglês | MEDLINE | ID: mdl-30428553

RESUMO

Regulation of appetite and food intake is partly regulated by N-acylethanolamine lipids oleoylethanolamide (OEA), stearoylethanolamide (SEA), and palmitoylethanolamide (PEA), which induce satiety through endogenous formation in the small intestine upon feeding, but also when orally or systemic administered. OEA, SEA, and PEA are present in human milk, and we hypothesized that the content of OEA, SEA, and PEA in mother's milk differed for infants being heavy (high weight-for-age Z-score (WAZ)) or light (low WAZ) at time of milk sample collection. Ultra-high performance liquid chromatography-mass spectrometry was used to determine the concentration of OEA, SEA, and PEA in milk samples collected four months postpartum from mothers to high (n = 50) or low (n = 50) WAZ infants. Associations between OEA, SEA, and PEA concentration and infant anthropometry at four months of age as well as growth from birth were investigated using linear and logistic regression analyses, adjusted for birth weight, early infant formula supplementation, and maternal pre-pregnancy body mass index. Mean OEA, SEA, and PEA concentrations were lower in the high compared to the low WAZ group (all p < 0.02), and a higher concentration of SEA was associated with lower anthropometric measures, e.g., triceps skinfold thickness (mm) (ß = -2.235, 95% CI = -4.04, -0.43, p = 0.016), and weight gain per day since birth (g) (ß = -8.169, 95% CI = -15.26, -1.08, p = 0.024). This raises the possibility, that the content of satiety factors OEA, SEA, and PEA in human milk may affect infant growth.


Assuntos
Peso Corporal , Endocanabinoides/metabolismo , Etanolaminas/metabolismo , Leite Humano/química , Ácidos Oleicos/metabolismo , Ácidos Palmíticos/metabolismo , Ácidos Esteáricos/metabolismo , Adulto , Envelhecimento , Aleitamento Materno , Estudos de Coortes , Dinamarca , Endocanabinoides/química , Etanolaminas/química , Feminino , Humanos , Lactente , Leite Humano/metabolismo , Ácidos Oleicos/química , Ácidos Palmíticos/química , Ácidos Esteáricos/química
10.
Proc Natl Acad Sci U S A ; 115(43): E10032-E10040, 2018 10 23.
Artigo em Inglês | MEDLINE | ID: mdl-30301806

RESUMO

Palmitoylethanolamide is a bioactive lipid that strongly alleviates pain and inflammation in animal models and in humans. Its signaling activity is terminated through degradation by N-acylethanolamine acid amidase (NAAA), a cysteine hydrolase expressed at high levels in immune cells. Pharmacological inhibitors of NAAA activity exert profound analgesic and antiinflammatory effects in rodent models, pointing to this protein as a potential target for therapeutic drug discovery. To facilitate these efforts and to better understand the molecular mechanism of action of NAAA, we determined crystal structures of this enzyme in various activation states and in complex with several ligands, including both a covalent and a reversible inhibitor. Self-proteolysis exposes the otherwise buried active site of NAAA to allow catalysis. Formation of a stable substrate- or inhibitor-binding site appears to be conformationally coupled to the interaction of a pair of hydrophobic helices in the enzyme with lipid membranes, resulting in the creation of a linear hydrophobic cavity near the active site that accommodates the ligand's acyl chain.


Assuntos
Amidoidrolases/metabolismo , Analgésicos/farmacologia , Animais , Domínio Catalítico/efeitos dos fármacos , Linhagem Celular , Descoberta de Drogas/métodos , Inibidores Enzimáticos/farmacologia , Etanolaminas/metabolismo , Humanos , Inflamação/metabolismo , Ligantes , Camundongos , Dor/tratamento farmacológico , Dor/metabolismo , Ácidos Palmíticos/metabolismo , Coelhos , Células Sf9 , Relação Estrutura-Atividade
11.
Sci Rep ; 8(1): 13734, 2018 09 13.
Artigo em Inglês | MEDLINE | ID: mdl-30214048

RESUMO

Understanding the epigenetic mechanisms underlying the progression of hepatic steatosis is important for identifying new therapeutic targets against nonalcoholic fatty liver disease (NAFLD). We investigated the functional role of histone demethylase JMJD2B in the pathologic regulation of hepatic steatosis. JMJD2B expression was markedly increased in HepG2 cells treated with palmitate and oleate or liver X receptor agonist T09013178 and in the liver of high-fat diet (HFD)-induced obese mice. Overexpression of JMJD2B using adenovirus in HepG2 cells stimulated the expression of peroxisome proliferator-activated receptor γ2 (PPARγ2) and its steatosis target genes associated with fatty acid uptake and lipid droplet formation, resulting in increased intracellular triglyceride (TG) accumulation. Conversely, knocking down JMJD2B using siRNA reversed JMJD2B-mediated effects in HepG2 cells. The JMJD2B-dependent upregulation of PPARγ2 was associated with the removal of di- and trimethylation of histone H3 lysine 9 on the promoter of PPARγ2. Furthermore, exogeneous expression of JMJD2B using adenovirus in mice resulted in hepatic steatosis when fed a HFD, which was accompanied with increased expression of hepatic PPARγ2 and its steatosis target genes. Together, our results provide novel insights into the pivotal role of JMJD2B in the development of hepatic steatosis through upregulation of PPARγ2 and steatosis target genes.


Assuntos
Fígado Gorduroso/genética , Histona Desmetilases com o Domínio Jumonji/genética , Hepatopatia Gordurosa não Alcoólica/genética , PPAR gama/genética , Animais , Dieta Hiperlipídica , Fígado Gorduroso/metabolismo , Fígado Gorduroso/patologia , Regulação da Expressão Gênica , Células Hep G2 , Humanos , Fígado/enzimologia , Fígado/metabolismo , Camundongos , Camundongos Obesos , Hepatopatia Gordurosa não Alcoólica/metabolismo , Hepatopatia Gordurosa não Alcoólica/patologia , Ácido Oleico/metabolismo , Ácidos Palmíticos/metabolismo , Triglicerídeos/genética
12.
Brain Behav Immun ; 74: 166-175, 2018 11.
Artigo em Inglês | MEDLINE | ID: mdl-30193877

RESUMO

Autism spectrum disorders (ASD) are a group of heterogeneous neurodevelopmental conditions characterized by impaired social interaction, and repetitive stereotyped behaviours. Interestingly, functional and inflammatory gastrointestinal diseases are often reported as a comorbidity in ASDs, indicating gut-brain axis as a novel emerging approach. Recently, a central role for peroxisome-proliferator activated receptor (PPAR)-α has been addressed in neurological functions, associated with the behaviour. Among endogenous lipids, palmitoylethanolamide (PEA), a PPAR-α agonist, has been extensively studied for its anti-inflammatory effects both at central and peripheral level. Based on this background, the aim of this study was to investigate the pharmacological effects of PEA on autistic-like behaviour of BTBR T+tf/J mice and to shed light on the contributing mechanisms. Our results showed that PEA reverted the altered behavioural phenotype of BTBR mice, and this effect was contingent to PPAR-α activation. Moreover, PEA was able to restore hippocampal BDNF signalling pathway, and improve mitochondrial dysfunction, both pathological aspects, known to be consistently associated with ASDs. Furthermore, PEA reduced the overall inflammatory state of BTBR mice, reducing the expression of pro-inflammatory cytokines at hippocampal, serum, and colonic level. The analysis of gut permeability and the expression of colonic tight junctions showed a reduction of leaky gut in PEA-treated BTBR mice. This finding together with PEA effect on gut microbiota composition suggests an involvement of microbiota-gut-brain axis. In conclusion, our results demonstrated a therapeutic potential of PEA in limiting ASD symptoms, through its pleiotropic mechanism of action, supporting neuroprotection, anti-inflammatory effects, and the modulation of gut-brain axis.


Assuntos
Transtorno do Espectro Autista/tratamento farmacológico , Transtorno do Espectro Autista/metabolismo , Etanolaminas/farmacologia , Ácidos Palmíticos/farmacologia , Animais , Transtorno Autístico/tratamento farmacológico , Transtorno Autístico/metabolismo , Encéfalo/metabolismo , Fator Neurotrófico Derivado do Encéfalo/metabolismo , Colo/metabolismo , Citocinas/metabolismo , Modelos Animais de Doenças , Etanolaminas/metabolismo , Microbioma Gastrointestinal/efeitos dos fármacos , Hipocampo/efeitos dos fármacos , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Mitocôndrias/efeitos dos fármacos , PPAR alfa/efeitos dos fármacos , PPAR alfa/metabolismo , Ácidos Palmíticos/metabolismo , Transdução de Sinais/efeitos dos fármacos
13.
Microbiol Immunol ; 62(8): 497-506, 2018 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-29932223

RESUMO

Lauroyltransferase gene (lpxL), Myristoyltransferase gene (lpxM) and palmitoyltransferase gene (crcA) of Escherichia coli BL21 were independently disrupted by the insertional mutations. The knockout mutant of two transferase genes (lpxL and crcA) produced lipid A with no lauric or palmitic acids and only a little amount of myristic acid. The mutant was susceptible to polymyxin B, but showed comparable growth with the wild-type strain at 30°C. The palmitoyltransferase gene from E. coli (crcA) or Salmonella (pagP) was amplified by PCR, cloned in pUC119, and transferred into the double-knockout mutant by transformation. The transformant contained palmitic acid in the lipid A, and recovered resistance to polymyxin B. Mass spectrometric analysis revealed that palmitic acid was linked to the hydroxyl group of 3-hydroxymyristic acid at C-2 position of proximal (reducing-end) glucosamine. LPS from the double-knockout mutant showed reduced IL-6-inducing activity to macrophage-like line cells compared to that of the wild-type strain, and the activity was only slightly restored by the introduction of palmitic acid to the lipid A. These results suggested that the introduction of one palmitic acid was enough to recover the integrity of the outer membrane, but not enough for the stimulation of macrophages.


Assuntos
Aciltransferases/genética , Proteínas de Escherichia coli/genética , Escherichia coli/genética , Lipídeo A/química , Lipídeo A/genética , Lipídeo A/metabolismo , Animais , Proteínas de Bactérias/genética , Escherichia coli/efeitos dos fármacos , Escherichia coli/crescimento & desenvolvimento , Proteínas de Escherichia coli/metabolismo , Técnicas de Inativação de Genes , Humanos , Interleucina-6/metabolismo , Ácidos Láuricos/metabolismo , Macrófagos/metabolismo , Camundongos , Testes de Sensibilidade Microbiana , Mutação , Ácido Mirístico/metabolismo , Ácidos Mirísticos/química , Ácidos Palmíticos/metabolismo , Polimixina B/farmacologia , Células RAW 264.7/efeitos dos fármacos , Salmonella/genética , Células U937/efeitos dos fármacos
14.
Gene ; 670: 123-129, 2018 Sep 05.
Artigo em Inglês | MEDLINE | ID: mdl-29787827

RESUMO

In the dairy industry, genetic variants have contributed to the improvement of milk production traits. Fatty acid elongase 6 (ELOVL6), which elongates saturated fatty acids (SFAs) and monounsaturated fatty acids (MUFAs), plays a distinct role in the balance of long-chain fatty acids composition in animals. ELOVL6 catalyzes the elongation of palmitic acids (C16:0) which is the most common saturated fatty acid found in animals and also an essential precursor to synthesize other long-chain fatty acids. However, the genetic variant research of bovine ELOVL6 on milk quality traits is still unknown. Therefore, our study aimed to detect the single nucleotide polymorphism (SNPs) of bovine ELOVL6 and explored the relationship between SNPs and milk quality traits including milk yield, fat content, protein content and somatic cell score. In this study, three SNPs, including SNP1 (g16379651A>G), SNP2 (g16458976A>G) and SNP3 (g16511290A>G), have been identified in intron 3 and 3'UTR regions of ELOVL6 in Chinese Holstein (CH) cows. Besides, the results of genetic diversity analysis, linkage disequilibrium and haplotype analysis indicated that these SNPs presented moderate polymorphisms which reflected relatively high genetic diversity. No strong linkage among these SNPs were detected in sampled population of cows. Moreover, the results of correlation analyses demonstrated that these SNPs of bovine ELOVL6 were significantly related to milk yield (P < 0.05). The SNP1 was also correlated with somatic cell score, whereas the SNP3 was associated with fat content. The 21 combined genotypes (diplotypes) were highly significantly correlated (P < 0.01) with milk yield. These results revealed that the genetic variants of bovine ELOVL6 influenced the milk production of CH cows. Hence, the three SNPs could be regarded as molecular markers in marker-assisted selection (MAS) of the dairy cow breeding.


Assuntos
Acetiltransferases/genética , Leite/metabolismo , Polimorfismo de Nucleotídeo Único , Regiões 3' não Traduzidas , Animais , Bovinos , Feminino , Haplótipos , Íntrons , Desequilíbrio de Ligação , Ácidos Palmíticos/metabolismo , Locos de Características Quantitativas
15.
Metab Eng ; 47: 414-422, 2018 05.
Artigo em Inglês | MEDLINE | ID: mdl-29719215

RESUMO

ω-Hydroxy palmitic acid (ω-HPA) is a valuable compound for an ingredient of artificially synthesized ceramides and an additive for lubricants and adhesives. Production of such a fatty acid derivative is limited by chemical catalysis, but plausible by biocatalysis. However, its low productivity issue, including formations of unsaturated fatty acid (UFA) byproducts in host cells, remains as a hurdle toward industrial biological processes. In this study, to achieve selective and high-level production of ω-HPA from glucose in Escherichia coli, FadR, a native transcriptional regulator of fatty acid metabolism, and its regulon were engineered. First, FadR was co-expressed with a thioesterase with a specificity toward palmitic acid production to enhance palmitic acid production yield, but a considerable quantity of UFAs was also produced. In order to avoid the UFA production caused by fadR overexpression, FadR regulon was rewired by i) mutating FadR consensus binding sites of fabA or fabB, ii) integrating fabZ into fabI operon, and iii) enhancing the strength of fabI promoter. This approach led to dramatic increases in both proportion (48.3-83.0%) and titer (377.8 mg/L to 675.8 mg/L) of palmitic acid, mainly due to the decrease in UFA synthesis. Introducing a fatty acid ω-hydroxylase, CYP153A35, into the engineered strain resulted in a highly selective production of ω-HPA (83.5 mg/L) accounting for 87.5% of total ω-hydroxy fatty acids. Furthermore, strategies, such as i) enhancement in CYP153A35 activity, ii) expression of a fatty acid transporter, iii) supplementation of triton X-100, and iv) separation of the ω-HPA synthetic pathway into two strains for a co-culture system, were applied and resulted in 401.0 mg/L of ω-HPA production. For such selective productions of palmitic acid and ω-HPA, the rewiring of FadR regulation in E. coli is a promising strategy to develop an industrial process with economical downstream processing.


Assuntos
Proteínas de Bactérias , Escherichia coli , Glucose , Ácidos Palmíticos/metabolismo , Regulon , Proteínas Repressoras , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Escherichia coli/genética , Escherichia coli/metabolismo , Glucose/genética , Glucose/metabolismo , Proteínas Repressoras/genética , Proteínas Repressoras/metabolismo
16.
J Neuroinflammation ; 15(1): 94, 2018 Mar 24.
Artigo em Inglês | MEDLINE | ID: mdl-29573741

RESUMO

BACKGROUND: Diarrhea is a severe complication in HIV-1-infected patients with Trans-activator of transcription (HIV-1 Tat) protein being recognized as a major underlying cause. Beside its direct enterotoxic effects, Tat protein has been recently shown to affect enteric glial cell (EGC) activity. EGCs regulate intestinal inflammatory responses by secreting pro-inflammatory molecules; nonetheless, they might also release immune-regulatory factors, as palmytoilethanolamide (PEA), which exerts anti-inflammatory effects by activating PPARα receptors. We aimed at clarifying whether EGCs are involved in HIV-1 Tat-induced diarrhea and if PEA exerts antidiarrheal activity. METHODS: Diarrhea was induced by intracolonic administration of HIV-1 Tat protein in rats at day 1. PEA alone or in the presence of peroxisome proliferator-activated receptor (PPAR) antagonists was given intraperitoneally from day 2 to day 7. S100B, iNOS, NF-kappaB, TLR4 and GFAP expression were evaluated in submucosal plexi, while S100B and NO levels were measured in EGC submucosal plexi lysates, respectively. To verify whether PEA effects were PPARα-mediated, PPARα-/- mice were also used. After 7 days from diarrhea induction, endogenous PEA levels were measured in submucosal plexi homogenates deriving from rats and PPARα-/- mice. RESULTS: HIV-1 Tat protein induced rapid onset diarrhea alongside with a significant activation of EGCs. Tat administration significantly increased all hallmarks of neuroinflammation by triggering TLR4 and NF-kappaB activation and S100B and iNOS expression. Endogenous PEA levels were increased following HIV-1 Tat exposure in both wildtype and knockout animals. In PPARα-/- mice, PEA displayed no effects. In wildtype rats, PEA, via PPARα-dependent mechanism, resulted in a significant antidiarrheal activity in parallel with marked reduction of EGC-sustained neuroinflammation. CONCLUSIONS: EGCs mediate HIV-1 Tat-induced diarrhea by sustaining the intestinal neuroinflammatory response. These effects are regulated by PEA through a selective PPARα-dependent mechanism. PEA might be considered as an adjuvant therapy in HIV-1-induced diarrhea.


Assuntos
Antivirais/uso terapêutico , Diarreia/induzido quimicamente , Diarreia/tratamento farmacológico , Etanolaminas/uso terapêutico , Neuroglia/efeitos dos fármacos , Ácidos Palmíticos/uso terapêutico , Produtos do Gene tat do Vírus da Imunodeficiência Humana/toxicidade , Anestésicos Locais/uso terapêutico , Animais , Modelos Animais de Doenças , Etanolaminas/metabolismo , Trato Gastrointestinal/patologia , Trato Gastrointestinal/virologia , Regulação Viral da Expressão Gênica/efeitos dos fármacos , Lidocaína/uso terapêutico , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , NF-kappa B/metabolismo , PPAR alfa/deficiência , PPAR alfa/genética , Ácidos Palmíticos/metabolismo , Ratos , Ratos Wistar , Subunidade beta da Proteína Ligante de Cálcio S100/metabolismo
17.
Sci Rep ; 8(1): 2416, 2018 02 05.
Artigo em Inglês | MEDLINE | ID: mdl-29403000

RESUMO

Enhancing endogenous cannabinoid (eCB) signaling has been considered as a potential strategy for the treatment of stress-related conditions. Fatty acid amide hydrolase (FAAH) represents the primary degradation enzyme of the eCB anandamide (AEA), oleoylethanolamide (OEA) and palmitoylethanolamide (PEA). This study describes a potent reversible FAAH inhibitor, SSR411298. The drug acts as a selective inhibitor of FAAH, which potently increases hippocampal levels of AEA, OEA and PEA in mice. Despite elevating eCB levels, SSR411298 did not mimic the interoceptive state or produce the behavioral side-effects (memory deficit and motor impairment) evoked by direct-acting cannabinoids. When SSR411298 was tested in models of anxiety, it only exerted clear anxiolytic-like effects under highly aversive conditions following exposure to a traumatic event, such as in the mouse defense test battery and social defeat procedure. Results from experiments in models of depression showed that SSR411298 produced robust antidepressant-like activity in the rat forced-swimming test and in the mouse chronic mild stress model, restoring notably the development of inadequate coping responses to chronic stress. This preclinical profile positions SSR411298 as a promising drug candidate to treat diseases such as post-traumatic stress disorder, which involves the development of maladaptive behaviors.


Assuntos
Amidoidrolases/antagonistas & inibidores , Ansiolíticos/farmacologia , Transtornos de Ansiedade/tratamento farmacológico , Agonistas de Receptores de Canabinoides/metabolismo , Carbamatos/farmacologia , Dioxanos/farmacologia , Inibidores Enzimáticos/farmacologia , Receptores de Canabinoides/genética , Estresse Psicológico/tratamento farmacológico , Doença Aguda , Amidoidrolases/genética , Amidoidrolases/metabolismo , Animais , Ansiolíticos/síntese química , Transtornos de Ansiedade/fisiopatologia , Ácidos Araquidônicos/metabolismo , Carbamatos/síntese química , Doença Crônica , Dioxanos/síntese química , Endocanabinoides/metabolismo , Inibidores Enzimáticos/síntese química , Etanolaminas/metabolismo , Feminino , Expressão Gênica , Masculino , Camundongos , Ácidos Oleicos/metabolismo , Ácidos Palmíticos/metabolismo , Alcamidas Poli-Insaturadas/metabolismo , Ratos Sprague-Dawley , Receptores de Canabinoides/metabolismo , Estresse Psicológico/fisiopatologia
18.
Redox Biol ; 15: 441-451, 2018 05.
Artigo em Inglês | MEDLINE | ID: mdl-29413957

RESUMO

Peripheral leukocytes induce blood-brain barrier (BBB) dysfunction through the release of cytotoxic mediators. These include hypochlorous acid (HOCl) that is formed via the myeloperoxidase-H2O2-chloride system of activated phagocytes. HOCl targets the endogenous pool of ether phospholipids (plasmalogens) generating chlorinated inflammatory mediators like e.g. 2-chlorohexadecanal and its conversion product 2-chlorohexadecanoic acid (2-ClHA). In the cerebrovasculature these compounds inflict damage to brain microvascular endothelial cells (BMVEC) that form the morphological basis of the BBB. To follow subcellular trafficking of 2-ClHA we synthesized a 'clickable' alkyne derivative (2-ClHyA) that phenocopied the biological activity of the parent compound. Confocal and superresolution structured illumination microscopy revealed accumulation of 2-ClHyA in the endoplasmic reticulum (ER) and mitochondria of human BMVEC (hCMEC/D3 cell line). 2-ClHA and its alkyne analogue interfered with protein palmitoylation, induced ER-stress markers, reduced the ER ATP content, and activated transcription and secretion of interleukin (IL)-6 as well as IL-8. 2-ClHA disrupted the mitochondrial membrane potential and induced procaspase-3 and PARP cleavage. The protein kinase R-like ER kinase (PERK) inhibitor GSK2606414 suppressed 2-ClHA-mediated activating transcription factor 4 synthesis and IL-6/8 secretion, but showed no effect on endothelial barrier dysfunction and cleavage of procaspase-3. Our data indicate that 2-ClHA induces potent lipotoxic responses in brain endothelial cells and could have implications in inflammation-induced BBB dysfunction.


Assuntos
Encéfalo/metabolismo , Células Endoteliais/metabolismo , Mitocôndrias/efeitos dos fármacos , Ácidos Palmíticos/efeitos adversos , Adenina/análogos & derivados , Adenina/farmacologia , Apoptose/efeitos dos fármacos , Barreira Hematoencefálica/efeitos dos fármacos , Barreira Hematoencefálica/patologia , Encéfalo/patologia , Linhagem Celular , Estresse do Retículo Endoplasmático/efeitos dos fármacos , Humanos , Indóis/farmacologia , Interleucina-8/genética , Leucócitos/metabolismo , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Mitocôndrias/patologia , Ácidos Palmíticos/metabolismo , Peroxidase/metabolismo , eIF-2 Quinase/genética
19.
Chem Rev ; 118(3): 919-988, 2018 02 14.
Artigo em Inglês | MEDLINE | ID: mdl-29292991

RESUMO

Protein lipidation, including cysteine prenylation, N-terminal glycine myristoylation, cysteine palmitoylation, and serine and lysine fatty acylation, occurs in many proteins in eukaryotic cells and regulates numerous biological pathways, such as membrane trafficking, protein secretion, signal transduction, and apoptosis. We provide a comprehensive review of protein lipidation, including descriptions of proteins known to be modified and the functions of the modifications, the enzymes that control them, and the tools and technologies developed to study them. We also highlight key questions about protein lipidation that remain to be answered, the challenges associated with answering such questions, and possible solutions to overcome these challenges.


Assuntos
Alquil e Aril Transferases/metabolismo , Proteínas/metabolismo , Alquil e Aril Transferases/antagonistas & inibidores , Alquil e Aril Transferases/química , Animais , Humanos , Cinética , Ácidos Mirísticos/metabolismo , Ácidos Palmíticos/metabolismo , Mapas de Interação de Proteínas , Prenilação de Proteína , Proteínas/química , Especificidade por Substrato
20.
Addict Biol ; 23(6): 1242-1250, 2018 11.
Artigo em Inglês | MEDLINE | ID: mdl-29178411

RESUMO

Alcohol binge drinking is a heavy pattern of alcohol consumption increasingly used by young people. In a previous study, we reported that young drinkers with a 2-year history of binge alcohol consumption had an overactivation of the innate immune system and peripheral inflammation when compared with controls. In the present study, we measured several biolipids that are fatty acid derivatives belonging to the acylethanolamide or 2-acylglycerol families in the plasma of the same subjects (n = 42; 20 men and 22 women). We found that during abstinence, alcohol binge drinkers had elevated plasma levels of oleoylethanolamide, palmitoleoylethanolamide, arachidonoylethanolamide, dihomo-γ-linolenoyl ethanolamide and linoleoyl ethanolamide, which positively correlated with changes in the mRNA expression of key inflammatory markers in peripheral blood mononuclear cells, such as toll-like receptors (TLR4), pro-inflammatory cytokines/chemokines interleukin-1 beta, interleukin-6 and monocyte chemoattractant protein-1, and cyclooxygenase-2. Additionally, plasma oleoylethanolamide positively correlated with plasma levels of high mobility group box-1, which is a danger-associated molecular pattern and an endogenous TLR4 agonist, specifically in female alcohol binge drinkers. No changes were observed in 2-acylglycerols in alcohol binge drinkers, although sex-related differences in these bioactive lipids as well as in palmitoleoylethanolamide and docosatetraenoylethanolamide levels were detected. These results extend the previous clinical findings observed in patients diagnosed with long-term alcohol use disorder to young users and suggest a prominent role for these lipids in the response to acute alcohol exposure.


Assuntos
Bebedeira/sangue , Endocanabinoides/metabolismo , Etanolaminas/metabolismo , Proteína HMGB1/metabolismo , Ácidos Oleicos/metabolismo , Ácidos Palmíticos/metabolismo , Antropometria , Biomarcadores/metabolismo , Estudos de Casos e Controles , Depressores do Sistema Nervoso Central/sangue , Depressores do Sistema Nervoso Central/metabolismo , Etanol/sangue , Etanol/metabolismo , Feminino , Glicerídeos/metabolismo , Humanos , Inflamação/metabolismo , Fígado/metabolismo , Masculino , Adulto Jovem
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