RESUMO
Foods enriched with insects can potentially prevent several health disorders, including cardiovascular diseases, by reducing inflammation and improving antioxidant status. In this study, Tenebrio molitor and Gryllus assimilis were selected to determine the effect on the development of atherosclerosis in ApoE/LDLR-/- mice. Animals were fed AIN-93G-based diets (control) with 10% Tenebrio molitor (TM) and 10% Gryllus assimilis (GA) for 8 weeks. The nutritional value as well as antioxidant activity of selected insects were determined. The lipid profile, liver enzyme activity, and the fatty acid composition of liver and adipose tissue of model mice were evaluated. Quantitative analysis of atherosclerotic lesions in the entire aorta was performed using the en face method, and for aortic roots, the cross-section method was used. The antioxidant status of the GA cricket was significantly higher compared to the TM larvae. The results showed that the area of atherosclerosis (en face method) was not significantly different between groups. Dietary GA reduced plaque formation in the aortic root; additionally, significant differences were observed in sections at 200 and 300 µm compared to other groups. Furthermore, liver enzyme ALT activity was lower in insect-fed groups compared to the control group. The finding suggests that a diet containing edible insect GA potentially prevents atherosclerotic plaque development in the aortic root, due to its high antioxidant activity.
Assuntos
Apolipoproteínas E , Aterosclerose , Receptores de LDL , Animais , Aterosclerose/patologia , Aterosclerose/metabolismo , Camundongos , Receptores de LDL/genética , Receptores de LDL/metabolismo , Apolipoproteínas E/deficiência , Apolipoproteínas E/genética , Insetos Comestíveis , Camundongos Knockout , Fígado/metabolismo , Fígado/patologia , Antioxidantes/metabolismo , Masculino , Tenebrio , Dieta , Aorta/patologia , Aorta/metabolismo , Modelos Animais de Doenças , Ração Animal , Placa Aterosclerótica/patologia , Placa Aterosclerótica/metabolismo , GryllidaeRESUMO
The aim of this study was to evaluate the anti-atherosclerotic effect of pomegranate seed oil as a source of conjugated linolenic acid (CLnA) (cis-9,trans-11,cis-13; punicic acid) compared to linolenic acid (LnA) and conjugated linoleic acid (CLA) (cis-9,trans-11) in apoE/LDLR-/- mice. In the LONG experiment, 10-week old mice were fed for the 18 weeks. In the SHORT experiment, 18-week old mice were fed for the 10 weeks. Diets were supplied with seed oils equivalent to an amount of 0.5% of studied fatty acids. In the SHORT experiment, plasma TCh and LDL+VLDL cholesterol levels were significantly decreased in animals fed CLnA and CLA compared to the Control. The expression of PPARα in liver was four-fold increased in CLnA group in the SHORT experiment, and as a consequence the expression of its target gene ACO was three-fold increased, whereas the liver's expression of SREBP-1 and FAS were decreased in CLnA mice only in the LONG experiment. Punicic acid and CLA isomers were determined in the adipose tissue and liver in animals receiving pomegranate seed oil. In both experiments, there were no effects on the area of atherosclerotic plaque in aortic roots. However, in the SHORT experiment, the area of atherosclerosis in the entire aorta in the CLA group compared to CLnA and LnA was significantly decreased. In conclusion, CLnA improved the lipid profile and affected the lipid metabolism gene expression, but did not have the impact on the development of atherosclerotic plaque in apoE/LDLR-/- mice.
Assuntos
Aterosclerose , Ácidos Linoleicos Conjugados , Placa Aterosclerótica , Punica granatum , Camundongos , Animais , Ácido alfa-Linolênico/farmacologia , Ácido alfa-Linolênico/metabolismo , Punica granatum/metabolismo , Metabolismo dos Lipídeos , Ácidos Linolênicos/farmacologia , Ácidos Linolênicos/metabolismo , Aterosclerose/tratamento farmacológico , Aterosclerose/genética , Óleos de Plantas/farmacologia , Apolipoproteínas E/genética , Apolipoproteínas E/metabolismo , Ácidos Linoleicos Conjugados/farmacologia , Ácidos Linoleicos Conjugados/metabolismoRESUMO
BACKGROUND: Genome-wide association studies have discovered a link between genetic variants on human chromosome 15q26.1 and increased coronary artery disease (CAD) susceptibility; however, the underlying pathobiological mechanism is unclear. This genetic locus contains the FES (FES proto-oncogene, tyrosine kinase) gene encoding a cytoplasmic protein-tyrosine kinase involved in the regulation of cell behavior. We investigated the effect of the 15q26.1 variants on FES expression and whether FES plays a role in atherosclerosis. METHODS AND RESULTS: Analyses of isogenic monocytic cell lines generated by CRISPR (clustered regularly interspaced short palindromic repeats)-mediated genome editing showed that monocytes with an engineered 15q26.1 CAD risk genotype had reduced FES expression. Small-interfering-RNA-mediated knockdown of FES promoted migration of monocytes and vascular smooth muscle cells. A phosphoproteomics analysis showed that FES knockdown altered phosphorylation of a number of proteins known to regulate cell migration. Single-cell RNA-sequencing revealed that in human atherosclerotic plaques, cells that expressed FES were predominately monocytes/macrophages, although several other cell types including smooth muscle cells also expressed FES. There was an association between the 15q26.1 CAD risk genotype and greater numbers of monocytes/macrophage in human atherosclerotic plaques. An animal model study demonstrated that Fes knockout increased atherosclerotic plaque size and within-plaque content of monocytes/macrophages and smooth muscle cells, in apolipoprotein E-deficient mice fed a high fat diet. CONCLUSIONS: We provide substantial evidence that the CAD risk variants at the 15q26.1 locus reduce FES expression in monocytes and that FES depletion results in larger atherosclerotic plaques with more monocytes/macrophages and smooth muscle cells. This study is the first demonstration that FES plays a protective role against atherosclerosis and suggests that enhancing FES activity could be a potentially novel therapeutic approach for CAD intervention.
Assuntos
Aterosclerose , Doença da Artéria Coronariana , Placa Aterosclerótica , Proteínas Proto-Oncogênicas c-fes , Animais , Humanos , Camundongos , Artérias/metabolismo , Aterosclerose/genética , Aterosclerose/metabolismo , Doença da Artéria Coronariana/genética , Doença da Artéria Coronariana/metabolismo , Estudo de Associação Genômica Ampla , Miócitos de Músculo Liso/metabolismo , Placa Aterosclerótica/genética , Placa Aterosclerótica/metabolismo , Proteínas Proto-Oncogênicas c-fes/genética , Proteínas Proto-Oncogênicas c-fes/metabolismoRESUMO
BACKGROUND AND PURPOSE: Vascular tone is regulated by the relative contractile state of vascular smooth muscle cells (VSMCs). Several integrins directly modulate VSMC contraction by regulating calcium influx through L-type voltage-gated Ca2+ channels (VGCCs). Genetic variants in ITGA9, which encodes the α9 subunit of integrin α9ß1, and SVEP1, a ligand for integrin α9ß1, associate with elevated blood pressure; however, neither SVEP1 nor integrin α9ß1 has reported roles in vasoregulation. We determined whether SVEP1 and integrin α9ß1 can regulate VSMC contraction. EXPERIMENTAL APPROACH: SVEP1 and integrin binding were confirmed by immunoprecipitation and cell binding assays. Human induced pluripotent stem cell-derived VSMCs were used in in vitro [Ca2+ ]i studies, and aortas from a Svep1+/- knockout mouse model were used in wire myography to measure vessel contraction. KEY RESULTS: We confirmed the ligation of SVEP1 to integrin α9ß1 and additionally found SVEP1 to directly bind to integrin α4ß1. Inhibition of SVEP1, integrin α4ß1 or α9ß1 significantly enhanced [Ca2+ ]i levels in isolated VSMCs to Gαq/11 -vasoconstrictors. This response was confirmed in whole vessels where a greater contraction to U46619 was seen in vessels from Svep1+/- mice compared to littermate controls or when integrin α4ß1 or α9ß1 was inhibited. Inhibition studies suggested that this effect was mediated via VGCCs, PKC and Rho A/Rho kinase dependent mechanisms. CONCLUSIONS AND IMPLICATIONS: Our studies reveal a novel role for SVEP1 and the integrins α4ß1 and α9ß1 in reducing VSMC contractility. This could provide an explanation for the genetic associations with blood pressure risk at the SVEP1 and ITGA9 loci.
Assuntos
Células-Tronco Pluripotentes Induzidas , Integrina alfa4beta1 , Ácido 15-Hidroxi-11 alfa,9 alfa-(epoximetano)prosta-5,13-dienoico , Animais , Cálcio/metabolismo , Moléculas de Adesão Celular , Humanos , Integrinas/genética , Integrinas/metabolismo , Ligantes , Camundongos , Vasoconstrição , Vasoconstritores , Quinases Associadas a rhoRESUMO
BACKGROUND: Dyslipidaemia is a major risk factor for atherosclerosis and cardiovascular diseases. The molecular mechanisms that translate dyslipidaemia into atherogenesis and reliable markers of its progression are yet to be fully elucidated. To address this issue, we conducted a comprehensive metabolomic and proteomic analysis in an experimental model of dyslipidaemia and in patients with familial hypercholesterolemia (FH). METHODS: Liquid chromatography/mass spectrometry (LC/MS) and immunoassays were used to find out blood alterations at metabolite and protein levels in dyslipidaemic ApoE-/-/LDLR-/- mice and in FH patients to evaluate their human relevance. RESULTS: We identified 15 metabolites (inhibitors and substrates of nitric oxide synthase (NOS), low-molecular-weight antioxidants (glutamine, taurine), homocysteine, methionine, 1-methylnicotinamide, alanine and hydroxyproline) and 9 proteins (C-reactive protein, proprotein convertase subtilisin/kexin type 9, apolipoprotein C-III, soluble intercellular adhesion molecule-1, angiotensinogen, paraoxonase-1, fetuin-B, vitamin K-dependent protein S and biglycan) that differentiated FH patients from healthy controls. Most of these changes were consistently found in dyslipidaemic mice and were further amplified if mice were fed an atherogenic (Western or low-carbohydrate, high-protein) diet. CONCLUSIONS: The alterations highlighted the involvement of an immune-inflammatory response system, oxidative stress, hyper-coagulation and impairment in the vascular function/regenerative capacity in response to dyslipidaemia that may also be directly engaged in development of atherosclerosis. Our study further identified potential biomarkers for an increased risk of atherosclerosis that may aid in clinical diagnosis or in the personalized treatment.
Assuntos
Aterosclerose , Dislipidemias , Hiperlipoproteinemia Tipo II , Animais , Aterosclerose/complicações , Dislipidemias/complicações , Humanos , Camundongos , Pró-Proteína Convertase 9 , Proteômica , Receptores de LDLRESUMO
Fiber optic Raman spectroscopy and Raman microscopy were used to investigate alterations in the aorta wall and the surrounding perivascular adipose tissue (PVAT) in the murine model of atherosclerosis (Apoe-/-/Ldlr-/- mice). Both abdominal and thoracic parts of the aorta were studied to account for the heterogenic chemical composition of aorta and its localization-dependent response in progression of atherosclerosis. The average Raman spectra obtained for both parts of aorta cross sections revealed that the chemical composition of intima-media layers along aorta remains relatively homogeneous while the lipid content in the adventitia layer markedly increases with decreasing distance to PVAT. Moreover, our results demonstrate that the increase of the lipid to protein ratio in the aorta wall correlates directly with the increased unsaturation level of lipids in PVAT and these changes occur only in the abdominal, but not in thoracic, aorta. In summary, distinct pathophysiological response in the aortic vascular wall could be uncovered by fiber optic Raman spectroscopy based on simple parameters detecting chemical contents of lipids in PVAT.
Assuntos
Aorta Abdominal/metabolismo , Aorta Torácica/metabolismo , Aterosclerose/metabolismo , Tecido Adiposo/metabolismo , Animais , Apolipoproteínas E/metabolismo , Lipídeos/química , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Receptores de LDL/metabolismo , Análise Espectral Raman/métodosRESUMO
Although, vitamin K2 displays vasoprotective effects, it is still not known whether K2 treatment improves endothelial function. In ApoE/LDLR-/- mice at the stage prior to atherosclerosis development, four-week treatment with K2-MK-7, given at a low dose (0.05â¯mg/kg), improved acetylcholine- and flow-induced, endothelium-dependent vasodilation in aorta or in femoral artery, as assessed by MRI in vivo. This effect was associated with an increased NO production, as evidenced by EPR measurements in ex vivo aorta. Treatment with higher doses of K2-MK-7 (0.5; 5â¯mg/kg) resulted in a dose-dependent increase in plasma K2-MK-7 and K2-MK-4 concentration, without further improvement in endothelial function. In ApoE/LDLR-/- mice with developed atherosclerotic plaques, treatment with a low (0.03â¯mg/kg) or high (10â¯mg/kg) dose of K2-MK-7 resulted in a similar degree of endothelium-dependent vasodilation improvement and increase in plasma nitrate concentration, what was not associated with changes in thrombin generation as measured by CAT. Both doses of K2-MK-7 also reduced media thickness in the brachiocephalic artery, but did not modify atherosclerotic plaque size. In conclusion, K2-MK-7 improves NO-dependent endothelial function in ApoE/LDLR-/- mice. This study, identifies the endothelial profile of the pharmacological activity of vitamin K2, which has not been previously described.
Assuntos
Aterosclerose/tratamento farmacológico , Endotélio Vascular/efeitos dos fármacos , Óxido Nítrico/metabolismo , Receptores de LDL/deficiência , Vasodilatação/efeitos dos fármacos , Vasodilatadores/farmacologia , Vitamina K 2/análogos & derivados , Fatores Etários , Animais , Aterosclerose/genética , Aterosclerose/metabolismo , Aterosclerose/fisiopatologia , Modelos Animais de Doenças , Progressão da Doença , Endotélio Vascular/metabolismo , Endotélio Vascular/fisiopatologia , Feminino , Masculino , Camundongos Knockout para ApoE , Placa Aterosclerótica , Receptores de LDL/genética , Transdução de Sinais , Fatores de Tempo , Vitamina K 2/farmacologiaRESUMO
This work presents the potential of vibrational spectroscopy, Vis and NIR Raman spectroscopy, Fourier transform infrared spectroscopy (FTIR) in reflection and transmission modes, and nano-FTIR microscopy to study the biochemical alterations in membranes of isolated and intact red blood cells (RBCs). The main goal was to propose the best spectroscopic method which enabled following biochemical alterations in the RBC membranes and then to translate this spectroscopic signature of degradation to in situ analysis of RBCs. Two models corresponding to two distinct cases of RBC membrane conditions were employed, and they were derived from healthy and young mice and mature mice with advanced atherosclerosis. It was shown that each technique provided essential information about biochemical alterations of the isolated membranes as well as membranes in the intact RBCs, which can be used in the development of a rapid and in situ analytical technology. Finally, we proposed that the combination of macro- and nanoprobing implemented in IR spectroscopy provided a wide chemical characterization of the RBC membranes, including alterations in lipid and protein fractions. This study also examined the effect of the sample preparation to determine destructive factors influencing a spectroscopic analysis of isolated membranes and intact RBCs derived from healthy and disease-affected mice.
Assuntos
Membrana Eritrocítica/química , Nanotecnologia/métodos , Espectroscopia de Infravermelho com Transformada de Fourier/métodos , Análise Espectral Raman/métodos , Animais , Colesterol/química , Esterificação , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Fosfolipídeos/químicaRESUMO
BACKGROUND: Genome-wide association studies have identified chromosome 14q32 as a locus for coronary artery disease. The disease-associated variants fall in a hitherto uncharacterized gene called HHIPL1 (hedgehog interacting protein-like 1), which encodes a sequence homolog of an antagonist of hedgehog signaling. The function of HHIPL1 and its role in atherosclerosis are unknown. METHODS: HHIPL1 cellular localization, interaction with sonic hedgehog (SHH), and influence on hedgehog signaling were tested. HHIPL1 expression was measured in coronary artery disease-relevant human cells, and protein localization was assessed in wild-type and Apoe-/- (apolipoprotein E deficient) mice. Human aortic smooth muscle cell phenotypes and hedgehog signaling were investigated after gene knockdown. Hhipl1-/- mice were generated and aortic smooth muscle cells collected for phenotypic analysis and assessment of hedgehog signaling activity. Hhipl1-/- mice were bred onto both the Apoe-/- and Ldlr-/- (low-density lipoprotein receptor deficient) knockout strains, and the extent of atherosclerosis was quantified after 12 weeks of high-fat diet. Cellular composition and collagen content of aortic plaques were assessed by immunohistochemistry. RESULTS: In vitro analyses revealed that HHIPL1 is a secreted protein that interacts with SHH and increases hedgehog signaling activity. HHIPL1 expression was detected in human smooth muscle cells and in smooth muscle within atherosclerotic plaques of Apoe-/- mice. The expression of Hhipl1 increased with disease progression in aortic roots of Apoe-/- mice. Proliferation and migration were reduced in Hhipl1 knockout mouse and HHIPL1 knockdown aortic smooth muscle cells, and hedgehog signaling was decreased in HHIPL1-deficient cells. Hhipl1 knockout caused a reduction of >50% in atherosclerosis burden on both Apoe-/- and Ldlr-/- knockout backgrounds, and lesions were characterized by reduced smooth muscle cell content. CONCLUSIONS: HHIPL1 is a secreted proatherogenic protein that enhances hedgehog signaling and regulates smooth muscle cell proliferation and migration. Inhibition of HHIPL1 protein function might offer a novel therapeutic strategy for coronary artery disease.
Assuntos
Aterosclerose/genética , Cromossomos Humanos Par 14/genética , Doença das Coronárias/genética , Proteínas Hedgehog/fisiologia , Peptídeos e Proteínas de Sinalização Intercelular/fisiologia , Animais , Aterosclerose/patologia , Divisão Celular , Movimento Celular , Células Cultivadas , Humanos , Peptídeos e Proteínas de Sinalização Intercelular/genética , Camundongos , Camundongos Knockout para ApoE , Miócitos de Músculo Liso/metabolismo , Placa Aterosclerótica/patologia , Receptores de LDL/deficiência , Transdução de SinaisRESUMO
Background The impairment of endothelium-dependent vasodilation, increased endothelial permeability, and glycocalyx degradation are all important pathophysiological components of endothelial dysfunction. However, it is still not clear whether in atherosclerosis, glycocalyx injury precedes other features of endothelial dysfunction or these events coincide. Methods and Results Herein, we demonstrate that in 4- to 8-week-old apolipoprotein E/low-density lipoprotein receptor-deficient mice, at the stage before development of atherosclerotic plaques, impaired acetylcholine-induced vasodilation, reduced NO production in aorta, and increased endothelial permeability were all observed; however, flow-mediated dilation in the femoral artery was fully preserved. In 4-week-old mice, glycocalyx coverage was reduced and endothelial stiffness was increased, whereas glycocalyx length was significantly decreased at 8 weeks of age. Early changes in endothelial function were also featured by increased plasma concentration of biomarkers of glycocalyx disruption (endocan), biomarkers of endothelial inflammation (soluble vascular cell adhesion molecule 1), increased vascular permeability (angiopoietin 2), and alterations in hemostasis (tissue plasminogen activator and plasminogen activator inhibitor 1). In 28-week-old mice, at the stage of advanced atherosclerotic plaque development, impaired NO production and nearly all other features of endothelial dysfunction were changed to a similar extent, compared with the preatherosclerotic plaque phase. The exceptions were the occurrence of acetylcholine-induced vasoconstriction in the aorta and brachiocephalic artery, impaired flow-mediated vasodilation in the femoral artery, and further reduction of glycocalyx length and coverage with a concomitant further increase in endothelial permeability. Conclusions In conclusion, even at the early stage before the development of atherosclerotic plaques, endothelial dysfunction is a complex multifactorial response that has not been previously appreciated.
Assuntos
Aorta Torácica/metabolismo , Endotélio Vascular/fisiopatologia , Glicocálix/metabolismo , Placa Aterosclerótica/metabolismo , Rigidez Vascular/fisiologia , Vasodilatação/fisiologia , Animais , Aorta Torácica/diagnóstico por imagem , Aorta Torácica/fisiopatologia , Apolipoproteínas E/deficiência , Tronco Braquiocefálico/diagnóstico por imagem , Tronco Braquiocefálico/metabolismo , Tronco Braquiocefálico/fisiopatologia , Modelos Animais de Doenças , Endotélio Vascular/metabolismo , Endotélio Vascular/patologia , Feminino , Imageamento Tridimensional , Imageamento por Ressonância Magnética , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Placa Aterosclerótica/diagnóstico , Placa Aterosclerótica/fisiopatologia , Receptores de LDL/deficiênciaRESUMO
Background: Nutritional recommendations emphasize the need to limit consumption of saturated fatty acids and to increase the intake of polyunsaturated fatty acids in the prevention of non-communicable chronic diseases, particularly cardiovascular diseases. Among the fatty acids with health-related effects on the body, conjugated fatty acids are mentioned (i.e. CLA). Objective: The current study was designed to determine the effects of conjugated linoleic acid (CLA) on serum lipid profile, glucose, liver enzymes activity (AST and ALT), malonic dialdehyde (MDA) as well as lipid hydroperoxide (LPO) concentrations in rats fed diet differing in type of dietary fat. Material and methods: Male Wistar rats were divided into six groups and fed the following diets: control AIN-93G diet contained soybean oil (O) and diets with modification of fat source: butter (B) and margarine (M). The experimental diets were supplemented with 1% of conjugated linoleic acid (O+CLA, B+CLA, M+CLA). After 21 days the blood was collected and lipid profile, glucose, liver enzymes, MDA as well as LPO were analyzed. Results: The dietary treatments had no significant effect on the body weight and liver weight of the animals. The concentrations of total cholesterol (TC) and LDL+VLDL cholesterol were unchanged. Both experimental factors (fat source and CLA) had a significant influence on the TAG and HDL levels. Margarine (M) significantly increased the TAG concentration, whereas CLA had a significant impact on the TAG reduction (M+CLA). Glucose level was significantly decreased in all groups fed diets supplemented with CLA. Serum ALT significantly increased in all CLA groups. Fat source had statistically significant influence on the MDA concentration. The LPO level was significantly elevated in all CLA groups. There was statistically significant interaction of experimental factors (fat source and CLA supplementation) on LPO level. Conclusions: Margarine had an adverse effect on the rat's lipid profile. However, in the group fed with margarine, the addition of CLA decreased the concentration of TAG. Regardless of the type of the dietary fat, CLA supplementation increased the level of LPO in the blood serum of animals.
Assuntos
Tecido Adiposo/efeitos dos fármacos , Ácidos Graxos/metabolismo , Ácidos Linoleicos Conjugados/farmacologia , Lipídeos/sangue , Estresse Oxidativo/efeitos dos fármacos , Animais , Masculino , Ratos , Ratos WistarRESUMO
Background: Caloric restriction (CR) leads to decrease metabolic intensity, which results in a reduction of oxygen consumption and the amount of free radicals. This can affect the function of the liver. Studies show that caloric restriction does not alter or significantly increase the enzyme activity associated with gluconeogenesis, but the effect was different according to the age of the model animals. Objective: The aim of the study was to determine the effect of caloric restriction on liver function in young and old ApoE/ LDLr-/- mice. Material and methods: Dietary experiments were performed on 2 and 5 month old male ApoE/LDLr-/- mice. Animals were divided into 3 experimental groups (n=6) and fed AIN'93G diet for 8 and 5 weeks, respectively. Control animals were fed ad libitum (AL) and housed in a colony cages. These animals were checked for dietary intake. The second group were also fed ad libitum but the animals were kept individually in cages (stress AL- sAL). Similarly to sAL group, the animals from the CR group were kept individually but received a 30% less diet compared to AL group. At the end of the experiment animals were euthanized and the blood, liver and adipose tissue have been collected. Alanine aminotransferase (ALT) as well as aspartate aminotransferase (AST) were measured in plasma. Fatty acid profile was evaluated (relative %) in adipose tissue (GC-MS). Liver's stetosis was assessed. Results were analyzed statistically (ANOVA, STATISTICA v.10.0). Results: CR ApoE/LDLr-/- mice showed significantly lower body weight compared to animals, both AL and sAL. There were no significant differences between ALT and AST in both younger and older animals. However, negative tendencies were more pronounced in younger animals. In young animals CR significantly increased liver weight compared to AL (4.14 vs 3.73g/100g). In adipose tissue fatty acid profile differed in CR mice compared to control in young animals. Conclusions: Caloric restriction did not affect liver enzymes in mice. Caloric restriction showed similar but not identical metabolic activity in young and old mice.
Assuntos
Alanina Transaminase/metabolismo , Fosfatase Alcalina/metabolismo , Restrição Calórica , Fígado/metabolismo , Animais , Peso Corporal , Hepatopatias/metabolismo , Camundongos , Camundongos Knockout , Tamanho do ÓrgãoRESUMO
The objective of the study was to determine the effects of pomegranate seed oil, used as a source of punicic acid (CLnA) in the diets of laying hens, on the physicochemical properties of eggs. Forty Isa Brown laying hens (26weeks old) were equally subjected to 4 dietary treatments (n=10) and fed a commercial layer diet supplying 2.5% sunflower oil (control) or three levels (0.5, 1.0 and 1.5%) of punicic acid in the diets. After 12weeks of feeding the hens, eggs collection began. Sixty eggs - randomly selected from each group - were analysed for physicochemical properties. Eggs naturally enriched with CLnA preserve their composition and conventional properties in most of the analysed parameters (including chemical composition, physical as well as organoleptic properties). Dietary CLnA had positive impact on the colour of the eggs' yolk, whereas the hardness of hard-boiled egg yolks was not affected. Additionally, increasing dietary CLnA led to an increase not only the CLnA concentrations, but also CLA in egg-yolk lipids.
Assuntos
Ração Animal/análise , Fenômenos Químicos/efeitos dos fármacos , Ovos/análise , Lythraceae , Óleos de Plantas/administração & dosagem , Sementes , Animais , Galinhas , Dieta/métodos , Gema de Ovo/química , Ácidos Graxos/análise , Feminino , Lipídeos/análise , Óleo de GirassolRESUMO
BACKGROUND: Cholesterol-dependent and independent mechanisms were proposed to explain anti-atherosclerotic action of statins in humans. However, their effects in murine models of atherosclerosis have not been consistently demonstrated. Here, we studied the effects of pravastatin on atherosclerosis in ApoE/LDLR-/- mice fed a control and atherogenic diet. METHODS: ApoE/LDLR-/- mice were fed a control (CHOW) or an atherogenic (Low Carbohydrate High Protein, LCHP) diet. Two doses of pravastatin (40mg/kg and 100mg/kg) were used. The anti-atherosclerotic effects of pravastatin in en face aorta, cross-sections of aortic roots and brachiocephalic artery (BCA) were analysed. The lipid profile was determined. Fourier Transform Infrared Spectroscopy followed by Fuzzy C-Means (FCM) clustering was used for the quantitative assessment of plaque composition. RESULTS: Treatment with pravastatin (100mg/kg) decreased total and LDL cholesterol only in the LCHP group, but displayed a pronounced anti-atherosclerotic effect in BCA and abdominal aorta. The anti-atherosclerotic effect of pravastatin (100mg/kg) in BCA was associated with significant alterations of the chemical plaque composition, including a fall in cholesterol and cholesterol esters contents independently on total cholesterol and LDL concentration in plasma. CONCLUSIONS: Pravastatin at high (100mg/kg), but not low dose displayed a pronounced anti-atherosclerotic effect in ApoE/LDLR-/- mice fed a CHOW or LCHP diet that was remarkable in BCA, visible in en face aorta, whereas it was not observed in aortic roots, suggesting that previous inconsistencies might have been due to the various sites of atherosclerotic plaque analysis.
Assuntos
Aorta/efeitos dos fármacos , Apolipoproteínas E/deficiência , Aterosclerose/tratamento farmacológico , Tronco Braquiocefálico/efeitos dos fármacos , Pravastatina/uso terapêutico , Receptores de LDL/deficiência , Animais , Anticolesterolemiantes/farmacologia , Anticolesterolemiantes/uso terapêutico , Aorta/metabolismo , Aorta/patologia , Aterosclerose/metabolismo , Aterosclerose/patologia , Tronco Braquiocefálico/metabolismo , Tronco Braquiocefálico/patologia , Feminino , Camundongos , Camundongos Knockout , Pravastatina/farmacologia , Resultado do TratamentoRESUMO
This study assessed the effects of individual conjugated linoleic acid isomers, c9t11-CLA and t10c12-CLA, on nonalcoholic fatty liver disease (NAFLD) and systemic endothelial dysfunction in rats fed for four weeks with control or high-fructose diet. The high-fructose diet hampered body weight gain (without influencing food intake), increased liver weight and glycogen storage in hepatocytes, upregulated expression of fatty acid synthase (FAS) and stearoyl-CoA desaturase-1 (SCD-1), and increased saturated fatty acid (SFA) content in the liver. Both CLA isomers prevented excessive accumulation of glycogen in the liver. Specifically, t10c12-CLA decreased concentration of serum triacylglycerols and LDL + VLDL cholesterol, increased HDL cholesterol, and affected liver lipid content and fatty acid composition by downregulation of liver SCD-1 and FAS expression. In turn, the c9t11-CLA decreased LDL+VLDL cholesterol in the control group and downregulated liver expression of FAS without significant effects on liver weight, lipid content, and fatty acid composition. In summary, feeding rats with a high-fructose diet resulted in increased liver glycogen storage, indicating the induction of gluconeogenesis despite simultaneous upregulation of genes involved in de novo lipogenesis. Although both CLA isomers (c9t11 and t10c12) display hepatoprotective activity, the hypolipemic action of the t10c12-CLA isomer proved to be more pronounced than that of c9t11-CLA.
Assuntos
Ácidos Linoleicos Conjugados/sangue , Glicogênio Hepático/metabolismo , Obesidade/sangue , Estearoil-CoA Dessaturase/biossíntese , Animais , Peso Corporal , Colesterol/sangue , Dieta , Frutose , Expressão Gênica/efeitos dos fármacos , Humanos , Isomerismo , Lipídeos/sangue , Glicogênio Hepático/química , Obesidade/patologia , Ratos , Estearoil-CoA Dessaturase/genética , Triglicerídeos/sangueRESUMO
The objective of this study was to compare effects of Western diet (WD) with low carbohydrate high protein (LCHP) diet on lipid metabolism, liver and kidney function in rats. Eighteen rats were randomly assigned to three experimental groups and fed for the next 2 months. The experimental diets were: Control (7% of soybean oil, 20% protein), WD (21% of butter, 20% protein), and LCHP (21% of butter and 52.4% protein) diet. The LCHP diet significantly decreased the body weight of the rats. Diet consumption was differentiated among groups, however significant changes were observed since third week of the experiment duration. Rats fed LCHP diet ate significantly less (25.2g/animal/day) than those from Control (30.2g/animal/day) and WD (27.8 g/animal/day) groups. Additionally, food efficiency ratio (FER) tended to decrease in LCHP fed rats. Serum homocysteine concentration significantly decreased in rats fed WD and LCHP diets. Liver weights were significantly higher in rats fed WD and LCHP diets. At the end of the experiment (2 months) the triacylglycerol (TAG) was significantly decreased in animals fed LCHP compared to WD. qRT-PCR showed that SCD-1 and FAS were decreased in LCHP fed rats, but WD diet increased expression of lipid metabolism genes. Rats receiving LCHP diet had two fold higher kidney weight and 54.5% higher creatinin level compared to Control and WD diets. In conclusion, LCHP diet decreased animal's body weight and decreased TAG in rat's serum. However, kidney damage in LCHP rats was observed.
Assuntos
Dieta Ocidental , Carboidratos da Dieta/administração & dosagem , Proteínas Alimentares/administração & dosagem , Animais , Peso Corporal , Ingestão de Alimentos , Ácido Graxo Sintases/genética , Expressão Gênica , Rim/patologia , Metabolismo dos Lipídeos , Fígado/metabolismo , Fígado/patologia , Masculino , Tamanho do Órgão , Ratos Wistar , Estearoil-CoA Dessaturase/genética , Triglicerídeos/sangueRESUMO
In terms of stereochemistry, catechins are divided into two groups: (-) epi forms (2R, 3R) and (-) forms (2S, 3R). Most of the catechins present in green tea are (-) epi forms (2R, 3R). Under the influence of high temperatures, in anaerobic conditions, as a result of epimerization the proportion of the (-) form (2S, 3R) increases. The data indicate that the presence of thermally modified catechins in the diet more efficiently reduces the development of atherosclerosis in apoE knockout mice than the presence of native catechins. The addition of the thermally modified formulations to the high-fat diet resulted in a reduction of the area of atherosclerotic lesions by about 28% (en face method) and 45% (cross-section method) compared to the group fed the high-fat diet without catechins. Furthermore, the body weight gain and plasma TBARS concentration in mice fed a diet with the addition of catechins depends on the degree of epimerization of catechins and decreases with increasing content of catechins belonging to the (-) form (2S, 3R). Moreover, plasma HDL cholesterol concentration in mice depends on catechins' stereoisomerism and increases along with the increasing content of catechins belonging to the (-) form (2S, 3R).
Assuntos
Doenças da Aorta/prevenção & controle , Aterosclerose/prevenção & controle , Catequina/farmacologia , Hipolipemiantes/farmacologia , Animais , Aorta/efeitos dos fármacos , Aorta/metabolismo , Aorta/patologia , Doenças da Aorta/sangue , Doenças da Aorta/genética , Doenças da Aorta/patologia , Apolipoproteínas E/deficiência , Apolipoproteínas E/genética , Aterosclerose/sangue , Aterosclerose/genética , Aterosclerose/patologia , Biomarcadores/sangue , Catequina/análogos & derivados , Catequina/química , HDL-Colesterol/sangue , Modelos Animais de Doenças , Hipolipemiantes/química , Masculino , Camundongos Knockout , Estrutura Molecular , Estereoisomerismo , Relação Estrutura-Atividade , Substâncias Reativas com Ácido Tiobarbitúrico/metabolismo , Triglicerídeos/sangue , Aumento de Peso/efeitos dos fármacosRESUMO
In this work the quantitative determination of atherosclerotic lesion area (ApoE/LDLR(-/-) mice) by FT-IR imaging is presented and validated by comparison with atherosclerotic lesion area determination by classic Oil Red O staining. Cluster analysis of FT-IR-based measurements in the 2800-3025 cm(-1) range allowed for quantitative analysis of the atherosclerosis plaque area, the results of which were highly correlated with those of Oil Red O histological staining (R(2) = 0.935). Moreover, a specific class obtained from a second cluster analysis of the aortic cross-section samples at different stages of disease progression (3, 4 and 6 months old) seemed to represent the macrophages (CD68) area within the atherosclerotic plaque.
Assuntos
Apolipoproteínas E/deficiência , Aterosclerose , Progressão da Doença , Placa Aterosclerótica/química , Receptores de LDL/deficiência , Espectroscopia de Infravermelho com Transformada de Fourier/métodos , Animais , Aterosclerose/metabolismo , Modelos Animais de Doenças , Feminino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Placa Aterosclerótica/metabolismoRESUMO
Although in apoE/LDLR(-/-) mice atherosclerotic plaques develop spontaneously, various atherogenic diets (e.g. Western diet) are frequently used to accelerate the disease in this model. The objective of this study was to compare the effects on atherosclerosis of Western diet and other types of high-fat, high cholesterol, hypertriglyceridemic diets with the effects of the low carbohydrate, high protein (LCHP) diet. 16-18 week old mice with pre-established atherosclerosis were assigned to experimental groups and fed for the next 10 weeks with control diet, margarine diet (margarine 7%), hypertrigliceridemic diet (fructose 62%), high-fat diet (Western diet), high cholesterol diet (egg yolk diet) or with LCHP diet. No differences in body weight were observed among experimental groups. Plasma cholesterol concentration was significantly increased in egg yolk diet- and LCHP diet-fed apoE/LDLR(-/-) mice as compared to other types of diets. Plasma concentration of triacylglycerols was significantly elevated in egg yolk diet- and LCHP diet-fed apoE/LDLR(-/-) mice. The area of atherosclerotic plaques in the aortic root was substantially increased in LCHP diet-fed mice as compared to other types of diets. Furthermore, in brachiocephalic arteries of LCHP diet-fed mice there was evidence of plaque rupture. In conclusion, the LCHP diet promoted atherosclerosis in apoE/LDLR(-/-) mice more intensively than classical Western diet and favored the development of unstable lesions.