RESUMEN
SCOPE: α-Cyclodextrin (α-CD), a cyclic polymer of glucose, has been shown to lower plasma cholesterol in animals and humans; however, its effect on atherosclerosis has not been previously described. METHODS AND RESULTS: apoE-knockout mice were fed either low-fat diet (LFD; 5.2% fat, w/w), or Western high fat diet (21.2% fat) containing either no additions (WD), 1.5% α-CD (WDA); 1.5% ß-CD (WDB); or 1.5% oligofructose-enriched inulin (WDI). Although plasma lipids were similar after 11 weeks on the WD vs. WDA diets, aortic atherosclerotic lesions were 65% less in mice on WDA compared to WD (P < 0.05), and similar to mice fed the LFD. No effect on atherosclerosis was observed for the other WD supplemented diets. By RNA-seq analysis of 16S rRNA, addition of α-CD to the WD resulted in significantly decreased cecal bacterial counts in genera Clostridium and Turicibacterium, and significantly increased Dehalobacteriaceae. At family level, Comamonadaceae significantly increased and Peptostreptococcaceae showed a negative trend. Several of these bacterial count changes correlated negatively with % atherosclerotic lesion and were associated with increased cecum weight and decreased plasma cholesterol levels. CONCLUSION: Addition of α-CD to the diet of apoE-knockout mice decreases atherosclerosis and is associated with changes in the gut flora.
Asunto(s)
Aterosclerosis/dietoterapia , Microbioma Gastrointestinal/efectos de los fármacos , Lípidos/sangre , alfa-Ciclodextrinas/farmacología , Animales , Aorta/efectos de los fármacos , Aorta/patología , Aterosclerosis/microbiología , Aterosclerosis/patología , Peso Corporal/efectos de los fármacos , Ciego/efectos de los fármacos , Ciego/microbiología , Dieta con Restricción de Grasas , Dieta Alta en Grasa/efectos adversos , Suplementos Dietéticos , Femenino , Microbioma Gastrointestinal/genética , Absorción Intestinal , Lípidos/farmacocinética , Ratones Noqueados para ApoE , alfa-Ciclodextrinas/metabolismo , beta-Ciclodextrinas/metabolismo , beta-Ciclodextrinas/farmacologíaRESUMEN
Aspirin (ASA) is known to alter the production of potent inflammatory lipid mediators, but whether it interacts with omega-3 fatty acids (FAs) from fish oil to affect atherosclerosis has not been determined. The goal was to investigate the impact of a fish oil-enriched diet alone and in combination with ASA on the production of lipid mediators and atherosclerosis. ApoE(-/-) female mice were fed for 13weeks one of the four following diets: omega-3 FA deficient (OD), omega-3 FA rich (OR) (1.8g omega-3 FAs/kg·diet per day), omega-3 FA rich plus ASA (ORA) (0.1g ASA/kg·diet per day) or an omega-3 FA deficient plus ASA (ODA) with supplement levels equivalent to human doses. Plasma lipids, atherosclerosis, markers of inflammation, hepatic gene expression and aortic lipid mediators were determined. Hepatic omega-3 FAs were markedly higher in OR (9.9-fold) and ORA (7-fold) groups. Mice in both OR and ORA groups had 40% less plasma cholesterol in very low-density lipoprotein-cholesterol and low-density lipoprotein fractions, but aortic plaque area formation was only significantly lower in the ORA group (5.5%) compared to the OD group (2.5%). Plasma PCSK9 protein levels were approximately 70% lower in the OR and ORA groups. Proinflammatory aortic lipid mediators were 50%-70% lower in the ODA group than in the OD group and more than 50% lower in the ORA group. In summary, less aortic plaque lesions and aortic proinflammatory lipid mediators were observed in mice on the fish oil diet plus ASA vs. just the fish oil diet.
Asunto(s)
Antiinflamatorios no Esteroideos/uso terapéutico , Aorta/efectos de los fármacos , Aspirina/uso terapéutico , Aterosclerosis/prevención & control , Células Progenitoras Endoteliales/efectos de los fármacos , Aceites de Pescado/uso terapéutico , Hígado/efectos de los fármacos , Animales , Antiinflamatorios no Esteroideos/efectos adversos , Aorta/inmunología , Aorta/metabolismo , Aorta/patología , Apolipoproteínas E/genética , Apolipoproteínas E/metabolismo , Aspirina/efectos adversos , Aterosclerosis/inmunología , Aterosclerosis/metabolismo , Aterosclerosis/patología , Biomarcadores/sangre , Biomarcadores/metabolismo , Proliferación Celular/efectos de los fármacos , Células Progenitoras Endoteliales/inmunología , Células Progenitoras Endoteliales/metabolismo , Células Progenitoras Endoteliales/patología , Ácidos Grasos Omega-3/efectos adversos , Ácidos Grasos Omega-3/sangre , Ácidos Grasos Omega-3/metabolismo , Ácidos Grasos Omega-3/uso terapéutico , Femenino , Aceites de Pescado/efectos adversos , Aceites de Pescado/metabolismo , Regulación de la Expresión Génica/efectos de los fármacos , Mediadores de Inflamación/sangre , Metabolismo de los Lípidos/efectos de los fármacos , Lípidos/sangre , Hígado/inmunología , Hígado/metabolismo , Hígado/patología , Ratones Noqueados , Proproteína Convertasa 9/sangre , Distribución Aleatoria , Aumento de Peso/efectos de los fármacosRESUMEN
Apolipoprotein C-II (apoC-II) is a cofactor for lipoprotein lipase, a plasma enzyme that hydrolyzes triglycerides (TGs). ApoC-II deficiency in humans results in hypertriglyceridemia. We used zinc finger nucleases to create Apoc2 mutant mice to investigate the use of C-II-a, a short apoC-II mimetic peptide, as a therapy for apoC-II deficiency. Mutant mice produced a form of apoC-II with an uncleaved signal peptide that preferentially binds high-density lipoproteins (HDLs) due to a 3-amino acid deletion at the signal peptide cleavage site. Homozygous Apoc2 mutant mice had increased plasma TG (757.5 ± 281.2 mg/dl) and low HDL cholesterol (31.4 ± 14.7 mg/dl) compared with wild-type mice (TG, 55.9 ± 13.3 mg/dl; HDL cholesterol, 55.9 ± 14.3 mg/dl). TGs were found in light (density < 1.063 g/ml) lipoproteins in the size range of very-low-density lipoprotein and chylomicron remnants (40-200 nm). Intravenous injection of C-II-a (0.2, 1, and 5 µmol/kg) reduced plasma TG in a dose-dependent manner, with a maximum decrease of 90% occurring 30 minutes after the high dose. Plasma TG did not return to baseline until 48 hours later. Similar results were found with subcutaneous or intramuscular injections. Plasma half-life of C-II-a is 1.33 ± 0.72 hours, indicating that C-II-a only acutely activates lipolysis, and the sustained TG reduction is due to the relatively slow rate of new TG-rich lipoprotein synthesis. In summary, we describe a novel mouse model of apoC-II deficiency and show that an apoC-II mimetic peptide can reverse the hypertriglyceridemia in these mice, and thus could be a potential new therapy for apoC-II deficiency.
Asunto(s)
Apolipoproteína C-II/genética , Materiales Biomiméticos/metabolismo , Hiperlipoproteinemia Tipo I/genética , Hipertrigliceridemia/genética , Mutación/genética , Fragmentos de Péptidos/genética , Secuencia de Aminoácidos , Animales , Femenino , Hiperlipoproteinemia Tipo I/sangre , Hipertrigliceridemia/sangre , Masculino , Ratones , Ratones Endogámicos C57BL , Datos de Secuencia Molecular , Embarazo , Triglicéridos/sangreRESUMEN
Lecithin-cholesterol acyltransferase (LCAT) is the major enzyme responsible for the esterification of free cholesterol on plasma lipoproteins, which is a key step in the reverse cholesterol transport pathway. The measurement of plasma LCAT activity not only is important in the diagnosis of patients with genetic or acquired LCAT deficiency but is also valuable in calculating cardiovascular risk, as well as in research studies of lipoprotein metabolism. In this chapter, we describe a convenient LCAT assay based on the use of an apoA-I mimetic peptide. The proteoliposome substrate used in this assay for LCAT is easily made with the peptide and can be stored by deep freezing without significant loss of activity.