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1.
Molecules ; 28(5)2023 Feb 28.
Artículo en Inglés | MEDLINE | ID: mdl-36903509

RESUMEN

Extra virgin olive oil (EVOO) has proven to yield a better health outcome than other saturated fats widely used in the Western diet, including a distinct dysbiosis-preventive modulation of gut microbiota. Besides its high content in unsaturated fatty acids, EVOO also has an unsaponifiable polyphenol-enriched fraction that is lost when undergoing a depurative process that gives place to refined olive oil (ROO). Comparing the effects of both oils on the intestinal microbiota of mice can help us determine which benefits of EVOO are due to the unsaturated fatty acids, which remain the same in both, and which benefits are a consequence of its minority compounds, mainly polyphenols. In this work, we study these variations after only six weeks of diet, when physiological changes are not appreciated yet but intestinal microbial alterations can already be detected. Some of these bacterial deviations correlate in multiple regression models with ulterior physiological values, at twelve weeks of diet, including systolic blood pressure. Comparison between the EVOO and ROO diets reveals that some of these correlations can be explained by the type of fat that is present in the diet, while in other cases, such as the genus Desulfovibrio, can be better understood if the antimicrobial role of the virgin olive oil polyphenols is considered.


Asunto(s)
Microbioma Gastrointestinal , Síndrome Metabólico , Ratones , Animales , Aceite de Oliva , Mantequilla , Dieta , Polifenoles
2.
PLoS One ; 17(8): e0271634, 2022.
Artículo en Inglés | MEDLINE | ID: mdl-35972974

RESUMEN

Butter and virgin olive oil (EVOO) are two fats differing in their degree of saturation and insaponifiable fraction. EVOO, enriched in polyphenols and other minority components, exerts a distinct effect on health. Using next generation sequencing, we have studied early and long-term effects of both types of fats on the intestinal microbiota of mice, finding significant differences between the two diets in the percentage of certain bacterial taxa, correlating with hormonal, physiological and metabolic parameters in the host. These correlations are not only concomitant, but most noticeably some of the changes detected in the microbial percentages at six weeks are correlating with changes in physiological values detected later, at twelve weeks. Desulfovibrionaceae/Desulfovibrio/D. sulfuricans stand out by presenting at six weeks a statistically significant higher percentage in the butter-fed mice with respect to the EVOO group, correlating with systolic blood pressure, food intake, water intake and insulin at twelve weeks. This not only suggests an early implication in the probability of developing altered physiological and biochemical responses later on in the host lifespan, but also opens the possibility of using this genus as a marker in the risk of suffering different pathologies in the future.


Asunto(s)
Microbioma Gastrointestinal , Síndrome Metabólico , Animales , Biomarcadores , Mantequilla , Dieta Alta en Grasa , Ratones , Aceite de Oliva/farmacología
3.
Nutrients ; 13(11)2021 Nov 04.
Artículo en Inglés | MEDLINE | ID: mdl-34836194

RESUMEN

(1) Background: Prolonged feeding with a high-fat diet (HFD) acts as a stressor by activating the functions of the hypothalamic-pituitary-adrenal gland (HPA) stress axis, accompanied of hypertension by inducing the renin-angiotensin-aldosterone system. Angiotensinases enzymes are regulatory aminopeptidases of angiotensin metabolism, which together with the dipeptidyl peptidase IV (DPP-IV), pyroglutamyl- and tyrosyl-aminopeptidase (pGluAP, TyrAP), participate in cognitive, stress, metabolic and cardiovascular functions. These functions appear to be modulated by the type of fat used in the diet. (2) Methods: To analyze a possible coordinated response of aminopeptidases, their activities were simultaneously determined in the hypothalamus, adenohypophysis and adrenal gland of adult male rats fed diets enriched with monounsaturated (standard diet (S diet) supplemented with 20% virgin olive oil; VOO diet) or saturated fatty acids (diet S supplemented with 20% butter and 0.1% cholesterol; Bch diet). Aminopeptidase activities were measured by fluorimetry using 2-Naphthylamine as substrates. (3) Results: the hypothalamus did not show differences in any of the experimental diets. In the pituitary, the Bch diet stimulated the renin-angiotensin system (RAS) by increasing certain angiotensinase activities (alanyl-, arginyl- and cystinyl-aminopeptidase) with respect to the S and VOO diets. DPP-IV activity was increased with the Bch diet, and TyrAP activity decrease with the VOO diet, having both a crucial role on stress and eating behavior. In the adrenal gland, both HFDs showed an increase in angiotensinase aspartyl-aminopeptidase. The interrelation of angiotensinases activities in the tissues were depending on the type of diet. In addition, correlations were shown between angiotensinases and aminopeptidases that regulate stress and eating behavior. (4) Conclusions: Taken together, these results support that the source of fat in the diet affects several peptidases activities in the HPA axis, which could be related to alterations in RAS, stress and feeding behavior.


Asunto(s)
Aminopeptidasas/efectos de los fármacos , Grasas de la Dieta/farmacología , Sistema Hipotálamo-Hipofisario/efectos de los fármacos , Sistema Hipófiso-Suprarrenal/efectos de los fármacos , Sistema Renina-Angiotensina/efectos de los fármacos , Glándulas Suprarrenales/metabolismo , Animales , Dieta Alta en Grasa/efectos adversos , Endopeptidasas/efectos de los fármacos , Ácidos Grasos/farmacología , Conducta Alimentaria/efectos de los fármacos , Hipotálamo/metabolismo , Masculino , Aceite de Oliva/farmacología , Adenohipófisis/metabolismo , Ratas , Ratas Wistar , Estrés Fisiológico/efectos de los fármacos
4.
Int J Mol Sci ; 22(10)2021 May 20.
Artículo en Inglés | MEDLINE | ID: mdl-34065436

RESUMEN

High saturated fat diets have been associated with the development of obesity and hypertension, along with other pathologies related to the metabolic syndrome. In contrast, the Mediterranean diet, characterized by its high content of monounsaturated fatty acids, has been proposed as a dietary factor capable of positively regulating cardiovascular function. These effects have been linked to changes in the local renal renin angiotensin system (RAS) and the activity of the sympathetic nervous system. The main goal of this study was to analyze the role of two dietary fat sources on aminopeptidases activities involved in local kidney RAS. Male Wistar rats (six months old) were fed during 24 weeks with three different diets: the standard diet (S), the standard diet supplemented with virgin olive oil (20%) (VOO), or the standard diet enriched with butter (20%) plus cholesterol (0.1%) (Bch). Kidney samples were separated in medulla and cortex for aminopeptidase activities (AP) assay. Urine samples were collected for routine analysis by chemical tests. Aminopeptidase activities were determined by fluorometric methods in soluble (sol) and membrane-bound (mb) fractions of renal tissue, using arylamide derivatives as substrates. After the experimental period, the systolic blood pressure (SBP) values were similar in standard and VOO animals, and significantly lower than in the Bch group. At the same time, a significant increase in GluAP and IRAP activities were found in renal medulla of Bch animals. However, in VOO group the increase of GluAP activity in renal medulla was lower, while AspAP activity decreased in the renal cortex. Furthermore, the VOO diet also affected other aminopeptidase activities, such as TyrAP and pGluAP, related to the regulation of the sympathetic nervous system and the metabolic rate. These results support the beneficial effect of VOO in the regulation of SBP through changes in local AP activities of the kidney.


Asunto(s)
Aminopeptidasas/metabolismo , Presión Sanguínea/efectos de los fármacos , Corteza Renal/efectos de los fármacos , Médula Renal/efectos de los fármacos , Aceite de Oliva/farmacología , Animales , Mantequilla , Colesterol/metabolismo , Dieta Mediterránea , Grasas de la Dieta/farmacología , Hipertensión/tratamiento farmacológico , Hipertensión/metabolismo , Corteza Renal/metabolismo , Médula Renal/metabolismo , Masculino , Obesidad/tratamiento farmacológico , Obesidad/metabolismo , Ratas , Ratas Wistar , Sistema Renina-Angiotensina/efectos de los fármacos
5.
Nutrients ; 13(2)2021 Jan 31.
Artículo en Inglés | MEDLINE | ID: mdl-33572630

RESUMEN

The brain renin-angiotensin system (RAS) has been recently involved in the homeostatic regulation of energy. Our goal was to analyse the influence of a diet rich in saturated fatty acids (butter) against one enriched in monounsaturated fatty acids (olive oil) on hypothalamic RAS, and their relationship with the metabolism of fatty acids. Increases in body weight and visceral fat, together with an increase in aminopeptidase A expression and reductions in AngII and AngIV were observed in the hypothalamus of animals fed with the butter diet. In this group, a marked reduction in the expression of genes related to lipid metabolism (LPL, CD36, and CPT-1) was observed in liver and muscle. No changes were found in terms of body weight, total visceral fat and the expression of hepatic genes related to fatty acid metabolism in the olive oil diet. The expressions of LPL and CD36 were reduced in the muscles, although the decrease was lower than in the butter diet. At the same time, the fasting levels of leptin were reduced, no changes were observed in the hypothalamic expression of aminopeptidase A and decreases were noted in the levels of AngII, AngIV and AngIII. These results support that the type of dietary fat is able to modify the hypothalamic profile of RAS and the body energy balance, related to changes in lipid metabolism.


Asunto(s)
Mantequilla , Hipotálamo/metabolismo , Metabolismo de los Lípidos , Aceite de Oliva/metabolismo , Sistema Renina-Angiotensina/fisiología , Angiotensina II/análogos & derivados , Angiotensina II/metabolismo , Angiotensina III/metabolismo , Animales , Peso Corporal , Antígenos CD36/metabolismo , Dieta Alta en Grasa , Metabolismo Energético , Ayuno/metabolismo , Expresión Génica , Glutamil Aminopeptidasa/metabolismo , Grasa Intraabdominal/crecimiento & desarrollo , Leptina/metabolismo , Metabolismo de los Lípidos/genética , Lipoproteína Lipasa/metabolismo , Hígado/metabolismo , Masculino , Ratones , Ratones Endogámicos ICR , Músculo Esquelético/metabolismo , ARN Mensajero/metabolismo , Reacción en Cadena en Tiempo Real de la Polimerasa , Aumento de Peso
6.
Front Mol Biosci ; 7: 94, 2020.
Artículo en Inglés | MEDLINE | ID: mdl-32500082

RESUMEN

Insulin-regulated aminopeptidase (IRAP, cystinyl aminopeptidase, CysAP) and aminopeptidase M (alanyl aminopeptidase, AlaAP) are closely related enzymes involved in cognitive, metabolic, and cardiovascular functions. These functions may be modulated by the type of fat used in the diet. In order to analyze a possible coordinated response of both enzymes we determined simultaneously their activities in frontal cortex, liver, and plasma of adult male rats fed diets enriched with fats differing in their percentages of saturated, mono or polyunsaturated fatty acids such as sesame, sunflower, fish, olive, Iberian lard, and coconut. The systolic blood pressure, food intake, body and liver weight as well as glucose and total cholesterol levels in plasma were measured. The type of fat in the diet influences the enzymatic activities depending on the enzyme and its location. These results suggest cognitive improvement properties for diets with predominance of polyunsaturated fatty acids. Physiological parameters such as systolic blood pressure, food intake, and biochemical factors such as cholesterol and glucose in plasma were also modified depending on the type of diet, supporting beneficial properties for diets rich in mono and polyunsaturated fatty acids. Inter-tissue correlations between the analyzed parameters were also modified depending on the type of diet. If the type of fat used in the diet modifies the behavior and relationship between CysAP and AlaAP in and between frontal cortex, liver and plasma, the functions in which they are involved could also be modified.

7.
Nutrients ; 12(3)2020 Feb 28.
Artículo en Inglés | MEDLINE | ID: mdl-32121057

RESUMEN

High-fat diets (HFD) have been widely associated with an increased risk of metabolic disorders and overweight. However, a high intake of sources that are rich in monounsaturated fatty acids has been suggested as a dietary agent that is able to positively influence energy metabolism and vascular function. The main objective of this study was to analyze the role of dietary fats on hepatic peptidases activities and metabolic disorders. Three diets: standard (S), HFD supplemented with virgin olive oil (VOO), and HFD supplemented with butter plus cholesterol (Bch), were administered over six months to male Wistar rats. Plasma and liver samples were collected for clinical biochemistry and aminopeptidase activities (AP) analysis. The expression of inducible nitric oxide synthase (iNOS) was also determined by Western blot in liver samples. The diet supplement with VOO did not induce obesity, in contrast to the Bch group. Though the VOO diet increased the time that was needed to return to the basal levels of plasma glucose, the fasting insulin/glucose ratio and HOMA2-%B index (a homeostasis model index of insulin secretion and valuation of ß-cell usefulness (% ß-cell secretion)) were improved. An increase of hepatic membrane-bound dipeptidyl-peptidase 4 (DPP4) activity was found only in VOO rats, even if no differences in fasting plasma glucagon-like peptide 1 (GLP-1) were obtained. Both HFDs induced changes in hepatic pyroglutamyl-AP in the soluble fraction, but only the Bch diet increased the soluble tyrosyl-AP. Angiotensinase activities that are implicated in the metabolism of angiotensin II (AngII) to AngIV increased in the VOO diet, which was in agreement with the higher activity of insulin-regulated-AP (IRAP) in this group. Otherwise, the diet that was enriched with butter increased soluble gamma-glutamyl transferase (GGT) and Leucyl-AP, iNOS expression in the liver, and plasma NO. In summary, VOO increased the hepatic activity of AP that were related to glucose metabolism (DPP4, angiotensinases, and IRAP). However, the Bch diet increased activities that are implicated in the control of food intake (Tyrosine-AP), the index of hepatic damage (Leucine-AP and GGT), and the expression of hepatic iNOS and plasma NO. Taken together, these results support that the source of fat in the diet affects several peptidases activities in the liver, which could be related to alterations in feeding behavior and glucose metabolism.


Asunto(s)
Dieta Alta en Grasa , Resistencia a la Insulina , Hígado/enzimología , Hígado/patología , Obesidad/enzimología , Péptido Hidrolasas/metabolismo , Animales , Dipeptidil Peptidasa 4/metabolismo , Ingestión de Energía , Conducta Alimentaria , Péptido 1 Similar al Glucagón/metabolismo , Prueba de Tolerancia a la Glucosa , Masculino , Óxido Nítrico/sangre , Óxido Nítrico Sintasa de Tipo II/metabolismo , Obesidad/sangre , Piroglutamil-Peptidasa I/metabolismo , Ratas Wistar , gamma-Glutamiltransferasa/metabolismo
8.
Antioxidants (Basel) ; 8(8)2019 Aug 02.
Artículo en Inglés | MEDLINE | ID: mdl-31382522

RESUMEN

The Mediterranean diet is associated with a low incidence of physiologic and metabolic non-communicable diseases such as hypertension, obesity, and insulin resistance. These chronic diseases are closely related to oxidative status, which is determined by the balance between oxidant and antioxidant levels. The Mediterranean diet is rich in foods with important antioxidant properties, such as fruits and extra virgin olive oil. The aim of this work was to establish the relationship between dietary patterns, the total intake of polyphenols, and the levels of 8-isoprostanes in urine, as a marker of lipid peroxidation, in a group of healthy Spanish women. The main sources of dietary polyphenols were fruits, vegetables, pulses, nuts, and extra virgin olive oil. There was a significant and positive correlation between the estimated intake of polyphenols, total polyphenols excreted in urine, adherence to the Mediterranean diet, and the intake of specific food groups. A positive correlation was established between the total polyphenols in urine and the intake of raw extra virgin olive oil. However, a negative correlation was established between the amount of 8-isoprostanes in urine, total intake of polyphenols, adherence to the Mediterranean diet, and the intake of fruits and nuts. These results indicate an association between oxidative status and the intake of foods that are typical of the Mediterranean diet, in healthy women. Furthermore, the results demonstrate the use of urine 8-isoprostanes as a marker of adherence to the Mediterranean diet.

9.
Vitam Horm ; 111: 105-129, 2019.
Artículo en Inglés | MEDLINE | ID: mdl-31421697

RESUMEN

After millennia of knowledge of opium, it was only recently that endogenous substances called opioids with similar properties to opium and derivatives were discovered. The first to be discovered were enkephalins. In addition to the regulation of their synthesis and expression of receptors, an important mechanism for the regulation of their functions carried out by multiple proteolytic enzymes acting at all levels of their structure is described. The action of such enzymes, known as enkephalinases, is also regulated by endogenous and exogenous factors which ultimately affect the control of the enkephalins's action. For therapeutic purposes, it is not only necessary to develop specific inhibitors but also to acquire a deep knowledge of the influence that such factors exert on their activities. This knowledge could help us to establish adapted therapeutic strategies in the treatment of pain or other processes in which enkephalinases are involved. In this chapter, some of these regulatory factors are discussed, such as regional and subcellular distribution, developmental changes, diurnal variations, hormonal influences, stress, dietary factors or interactions with other neurotransmitters.


Asunto(s)
Neprilisina/metabolismo , Animales , Encéfalo/crecimiento & desarrollo , Encéfalo/ultraestructura , Química Encefálica/fisiología , Ritmo Circadiano/fisiología , Dieta , Sistema Endocrino/fisiología , Encefalinas/fisiología , Femenino , Homeostasis , Humanos , Masculino , Neprilisina/análisis , Manejo del Dolor/métodos , Estrés Psicológico/enzimología , Fracciones Subcelulares/química , Distribución Tisular
10.
Microorganisms ; 7(2)2019 Feb 23.
Artículo en Inglés | MEDLINE | ID: mdl-30813410

RESUMEN

Extra virgin olive oil (EVOO) has been reported to have a distinct influence on gut microbiota in comparison to other fats, with its physiological benefits widely studied. However, a large proportion of the population consumes olive oil after a depurative process that not only mellows its taste, but also deprives it of polyphenols and other minority components. In this study, we compare the influence on the intestinal microbiota of a diet high in this refined olive oil (ROO) with other fat-enriched diets. Swiss Webster mice were fed standard or a high-fat diet enriched with EVOO, ROO, or butter (BT). Physiological parameters were also evaluated. At the end of the feeding period, DNA was extracted from feces and the 16S rRNA was pyrosequenced. The group fed ROO behaved differently to the EVOO group in half the families with statistically significant differences among the diets, with higher comparative levels in three families-Desulfovibrionaceae, Spiroplasmataceae, and Helicobacteraceae-correlating with total cholesterol. These results are again indicative of a link between specific diets, certain physiological parameters and the prevalence of some taxa, but also support the possibility that polyphenols and minor components of EVOO are involved in some of the proposed effects of this fat through the modulation of the intestinal microbiota.

11.
Neuro Endocrinol Lett ; 39(4): 277-280, 2018 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-30531704

RESUMEN

OBJECTIVES: Enkephalins functions are partly modulated by enkephalinases especially membrane-bound alanyl aminopeptidase (EC-3.4.11.2) considered as the major enkephalin-degrading enzyme in brain. The analysis of its activity in standard and non-standard light/dark conditions may help the understanding of the regulatory mechanisms of enkephalins. METHODS: Enkephalinase activity was determined fluorometrically, using an arylamide derivative as substrate, in hypothalamus and pituitary of adult male rats, in a standard 12:12 h light/dark cycle; samples were collected at 10:00 h, 13:00 h and 16:00 h of the light period (light on from 7:00 to 19:00 h) and at 22:00 h, 01:00 h and 04:00 h of the dark one (light off from 19:00 h to 07:00 h). For comparison, the enzymatic activity was also measured in the same locations at the same time-points but under constant light conditions. RESULTS: In standard light/dark conditions, the results demonstrated an opposite daily rhythm in hypothalamus and pituitary. While the highest levels of AlaAP or enkephalinase activities were measured in hypothalamus during the dark period, they were the highest in the pituitary during the light one. In contrast, the lowest levels of activity were observed in the light period in the hypothalamus whereas they were in the dark one in the pituitary. A similar pattern was observed under constant light. The differences were however higher in hypothalamus and lower, but still significant, in pituitary. CONCLUSION: These results may reflect the behaviour of the endogenous substrates of enkephalinase and consequently be involved in their functions. This observation may affect the chronotherapeutic strategies.


Asunto(s)
Hipotálamo/enzimología , Luz , Neprilisina/metabolismo , Fotoperiodo , Hipófisis/enzimología , Animales , Masculino , Ratas
12.
Molecules ; 23(7)2018 Jun 28.
Artículo en Inglés | MEDLINE | ID: mdl-29958403

RESUMEN

BACKGROUND: Glucagon-Like Peptide-1 (GLP-1) is hydrolyzed by Dipeptidyl-Peptidase 4 (DPP4), and several studies suggest that both GLP-1 and DPP4 inhibitors have potentially beneficial effects on cardiovascular risks. The objective of this study was to analyze the differences between plasma GLP-1 and DPP4 activity in male and female patients with metabolic syndrome, and its relationship with physiological and metabolic parameters. The study included 25 apparently healthy Controls (C) and 21 Metabolic Syndrome patients (MS). Anthropometric indices, cardiovascular risk-score, and Mediterranean Diet Adherence (AMeDit) were evaluated. Fasting glucose, glycosylated hemoglobin (HbA1c), and insulin were measured. Insulin, GLP-1, and plasma DPP4 were determined within the first 30-min postprandial period. Body-Mass-Index was significantly higher, and AMeDit was significantly lower, but only in MS women. However, fasting glucose, HbA1c, and postprandial insulin were significantly higher in MS men, but not in MS women. Postprandial GLP-1 levels were lower in C men than in C women. Interestingly, in comparison with controls, we found significant lower levels of plasma DPP4 in MS-women only. Moreover, negative lineal regressions were established between DPP4 activity with waist-to-hip ratio and cardiovascular risk-score, and positive lineal regression with AMeDit. These results indicate gender differences in the behavior of GLP-1 and DPP4 activity in MS, which could be relevant for its treatment with GLP-1 analogues and DPP4 inhibitors.


Asunto(s)
Dieta , Dipeptidil Peptidasa 4/sangre , Péptido 1 Similar al Glucagón/sangre , Síndrome Metabólico/sangre , Femenino , Humanos , Insulina/sangre , Masculino , Proyectos Piloto , Factores Sexuales
13.
PLoS One ; 13(1): e0190368, 2018.
Artículo en Inglés | MEDLINE | ID: mdl-29293629

RESUMEN

The type of fat in the diet determinates the characteristics of gut microbiota, exerting a major role in the development of metabolic syndrome. We hypothesize that a diet enriched with extra virgin olive oil (EVOO) has a distinctive effect on the intestinal microbiome in comparison with an enriched butter diet (BT) and this effect is related to the physiological benefits exerted by EVOO. Swiss Webster mice were fed standard (SD) or two high fat diets enriched with EVOO or butter. Hormonal, physiological and metabolic parameters were evaluated. At the end of the feeding period, DNA was extracted from faeces and the 16S rRNA genes were pyrosequenced. Among the main significant differences found, BT triggered the highest values of systolic blood pressure, correlating positively with the percentage of Desulfovibrio sequences in faeces, which in turn showed significantly higher values in BT than in EVOO. EVOO had the lowest values of plasmatic insulin, correlating inversely with Desulfovibrio, and had the lowest plasmatic values of leptin which correlated inversely with Sutterellaceae, Marispirillum and Mucilaginibacter dageonensis, the three showing significantly higher percentages in EVOO. The lowest total cholesterol levels in plasma were detected in SD, correlating positively with Prevotella and Fusicatenibacter, both taxa with significantly greater presence in SD. These results may be indicative of a link between specific diets, certain physiological parameters and the prevalence of some taxa, supporting the possibility that in some of the proposed effects of virgin olive oil the modulation of intestinal microbiota could be involved.


Asunto(s)
Mantequilla , Dieta , Modelos Animales de Enfermedad , Microbioma Gastrointestinal , Síndrome Metabólico/fisiopatología , Aceite de Oliva , Animales , Heces/microbiología , Masculino , Ratones , ARN Ribosómico 16S/genética
14.
Int J Mol Sci ; 18(8)2017 Aug 04.
Artículo en Inglés | MEDLINE | ID: mdl-28777292

RESUMEN

The aim of the present work was to improve our knowledge on the mechanisms underlying the beneficial or deleterious effects on testicular function of the so-called Mediterranean and Western diet by analyzing glutamyl aminopeptidase (GluAP), gamma glutamyl transpeptidase (GGT) and dipeptidyl peptidase IV (DPP IV) activities in testis, as enzymes involved in testicular function. Male Wistar rats (6 months old) were fed for 24 weeks with three different diets: standard (S), an S diet supplemented with virgin-olive-oil (20%) (VOO), or a S diet enriched with butter (20%) plus cholesterol (0.1%) (Bch). At the end of the experimental period, plasma lipid profiled (total triglycerides, total cholesterol and cholesterol fractions (HDL, LDL and VDL)) were measured. Enzymatic activities were determined by fluorimetric methods in soluble (sol) and membrane-bound (mb) fractions of testicular tissue using arylamide derivatives as substrates. Results indicated an increase in plasmatic triglycerides, total cholesterol, LDL and VLDL in Bch. A significant increase of mb GluAP and GGT activities was also found in this diet in comparison with the other two diets. Furthermore, significant and positive correlations were established between these activities and plasma triglycerides and/or total cholesterol. These results support a role for testicular GluAP and GGT activities in the effects of saturated fat (Western diet) on testicular functions. In contrast, VOO increased sol DPP IV activity in comparison with the other two diets, which support a role for this activity in the effects of monounsaturated fat (Mediterranean diet) on testicular function. The present results strongly support the influence of fatty acids and cholesterol on testicular GluAP and GGT activities and also provide support that the reported beneficial influence of the Mediterranean diet in male fertility may be mediated in part by an increase of testicular sol DPP IV activity.


Asunto(s)
Mantequilla , Colesterol/farmacología , Grasas Insaturadas en la Dieta/farmacología , Aceite de Oliva/farmacología , Testículo/enzimología , Animales , Peso Corporal/efectos de los fármacos , Colesterol/sangre , Dipeptidil Peptidasa 4/metabolismo , Glutamil Aminopeptidasa/metabolismo , Masculino , Ratas Wistar , Solubilidad , Testículo/efectos de los fármacos , gamma-Glutamiltransferasa/metabolismo
15.
PLoS One ; 12(4): e0175462, 2017.
Artículo en Inglés | MEDLINE | ID: mdl-28399178

RESUMEN

PURPOSE: The aim of this work was to investigate if the content of glutamyl aminopeptidase (GluAp) in microvesicular and exosomal fractions of urine is related with renal dysfunction in cisplatin-treated rats. METHODS: Urine samples were collected 24 hours after injection of cisplatin (7 mg/kg, n = 10) or saline serum (n = 10), and they were subjected to differential centrifugation at 1.000, 17.000 and 200.000 g to obtain microvesicular and exosomal fractions. GluAp was measured with a commercial ELISA kit in both fractions. Serum creatinine (SCr) and body weight were measured 15 days after treatment. We analyzed if early excretion of GluAp in microsomal and exosomal fractions was correlated with final SCr and body weight increase. In a second experiment, enzymatic activities of GluAp and alanyl aminopeptidase (AlaAp) in urine, microvesicular and exosomal fractions were measured three days after injection. We analyzed the correlation of both markers with SCr determined at this point. Finally, we studied the expression of GluAp and extracellular vesicles markers Alix and tumor susceptibility gene (TSG101) in both fractions by immunoblotting. RESULTS: GluAp excretion was increased in all fractions of urine after cisplatin treatment, even if data were normalized per mg of creatinine, per body weight or per total protein content of each fraction. We found significant predictive correlations with SCr concentration, and inverse correlations with body weight increase determined 15 days later. Three days after injection, aminopeptidasic activities were markedly increased in all fractions of urine in cisplatin-treated rats. The highest correlation coefficient with SCr was found for GluAp in microvesicular fraction. Increase of GluAp in microvesicular and exosomal fractions from cisplatin-treated rats was confirmed by immunoblotting. Alix and TSG101 showed different patterns of expression in each fraction. CONCLUSIONS: Determination of GluAp content or its enzymatic activity in microvesicular and exosomal fractions of urine is an early and predictive biomarker of renal dysfunction in cisplatin-induced nephrotoxicity. Measurement of GluAp in these fractions can serve to detect proximal tubular damage independently of glomerular filtration status.


Asunto(s)
Cisplatino/administración & dosificación , Exosomas/enzimología , Glutamil Aminopeptidasa/orina , Riñón/efectos de los fármacos , Animales , Cisplatino/toxicidad , Riñón/enzimología , Riñón/fisiopatología , Masculino , Ratas , Ratas Wistar
16.
Acta Neuropsychiatr ; 28(4): 239-43, 2016 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-26882874

RESUMEN

OBJECTIVE: To evaluate the influence of acute restraint stress (ARS) on plasma enkephalinase and oxytocinase activities. ARS modifies basal activities in cortico-limbic regions of rats and induces changes in the correlations observed between these regions. The interactions between plasma and cortico-limbic activities will be also evaluated. METHODS: Enkephalinase (AlaAP and LeuAP) and oxytocinase (P-LeuAP) activities were fluorometrically determined in plasma of control and stressed rats using aminoacyl-ß-naphthylamides (aaNNap), AlaNNap and LeuNNap as substrates. RESULTS: No differences in enzymatic activities were observed between control and stressed animals in plasma. In contrast, highly significant positive and negative correlations between plasma and cortico-limbic regions were demonstrated in controls. Stress conditions significantly alter the pattern of these correlations. CONCLUSION: The present results clearly support a connection between plasma and brain involving certain neuropeptidase activities that change under stress conditions.


Asunto(s)
Encéfalo/fisiología , Cistinil Aminopeptidasa/sangre , Neprilisina/sangre , Estrés Fisiológico/fisiología , Animales , Estudios de Casos y Controles , Masculino , Distribución Aleatoria , Ratas , Ratas Wistar , Restricción Física
17.
Life Sci ; 134: 73-8, 2015 Aug 01.
Artículo en Inglés | MEDLINE | ID: mdl-26006037

RESUMEN

AIMS: To better understand the functional role of soluble (Sol) and membrane-bound (MB) cystinyl-aminopeptidase (CysAP) activities, we studied differentially their organ distribution in adult male Wistar-Kyoto (WKY) and spontaneously hypertensive rats (SHR)with or without treatment with captopril.We searched for a possible tissue-specific association of CysAP with water balance and blood pressure. MAIN METHODS: We used twenty WKY rats distributed in ten controls and ten captopril-treated, and sixteen SHR divided in eight controls and eight captopril-treated. Captopril (100 mg/kg/day) was administered in drinking water for 4 weeks. Systolic blood pressure, water intake and diuresis were measured individually. CysAP was assayed fluorometrically using L-cystine-di-ß-naphthylamide as substrate. KEY FINDINGS: Sol or MB activities were generally higher in SHR compared to WKY notably in hypothalamus and kidney than in the other tissues. Captopril mainly decreased CysAP in SHR whereas it increased in WKY. The distribution of Sol CysAP was more homogeneous among tissues ofWKY than SHR. In contrast, the distribution of MB CysAP was more heterogeneous than Sol CysAP in both WKY and SHR. This suggests that MB CysAP activity acts in a more tissue-specific manner than Sol CysAP. The majority of the significant correlations between tissue activities and the measured physiological parameters were observed mostly in renal medulla and hypothalamus. SIGNIFICANCE: Sol and MB CysAP activities, acting separately or in concert and mainly in renal medulla, regulate the function of their susceptible endogenous substrates, and may participate meaningfully in the control of blood pressure and fluid balance.


Asunto(s)
Presión Sanguínea/fisiología , Cistinil Aminopeptidasa/metabolismo , Hipocampo/enzimología , Médula Renal/enzimología , Equilibrio Hidroelectrolítico/fisiología , Animales , Antihipertensivos/farmacología , Presión Sanguínea/efectos de los fármacos , Captopril/farmacología , Masculino , Especificidad de Órganos/efectos de los fármacos , Ratas , Ratas Endogámicas SHR , Ratas Endogámicas WKY , Equilibrio Hidroelectrolítico/efectos de los fármacos
18.
Planta Med ; 81(8): 664-9, 2015 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-25389059

RESUMEN

High-fat diets are associated with the development of cardiovascular diseases. The efficacy of the current strategies of treatment is still not entirely satisfactory, and new approaches are being considered. To analyze the beneficial effects of extra virgin olive oil as a major component of the Mediterranean diet, we studied systolic blood pressure and angiotensinase activities, since this enzyme is involved in the metabolism of angiotensins, in the kidney of hypertensive rats fed during 12 weeks with a diet enriched with extra virgin olive oil compared with a standard diet. As a reflex of oxidative stress, 8-isoprostanes and nitric oxide were quantified in urine. Results demonstrated a progressive increase in systolic blood pressure until the end of the feeding period in both groups. However, this increase was delayed in the extra virgin olive oil group until week six, with the systolic blood pressure being always lower in this group. Nitric oxide and 8-isoprostanes were lower in the extra virgin olive oil group. While we can deduce a higher formation of angiotensin 2-10 in the renal cortex, a higher availability of angiotensin II may be presumed in the renal medulla of animals fed an extra virgin olive oil diet than in animals fed a standard diet. Our results support the beneficial influence of extra virgin olive oil on cardiovascular function and suggest that the Mediterranean diet may be beneficial in itself but it may also be an effective tool in the treatment of hypertension.


Asunto(s)
Enfermedades Cardiovasculares/tratamiento farmacológico , Endopeptidasas/efectos de los fármacos , Hipertensión/tratamiento farmacológico , Aceite de Oliva/farmacología , Animales , Presión Sanguínea/efectos de los fármacos , Dieta Alta en Grasa/efectos adversos , Dieta Mediterránea , Modelos Animales de Enfermedad , Endopeptidasas/metabolismo , Riñón/enzimología , Masculino , Estrés Oxidativo/efectos de los fármacos , Ratas , Ratas Endogámicas SHR
19.
J Endocrinol ; 220(3): 333-43, 2014 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-24389591

RESUMEN

The metabolic action of oxytocin has recently been intensively studied to assess the ability of the peptide to regulate energy homeostasis. Despite the obvious weight-reducing effect of oxytocin observed in experimental studies, plasma oxytocin levels were found to be unchanged or even elevated in human obesity. The aim of our study was to evaluate the changes in the oxytocin system in Zucker rats, an animal model closely mirroring morbid obesity in humans. Plasma oxytocin levels were measured in obese Zucker rats and lean controls by enzyme immunoassay after plasma extraction. The expression of oxytocin and oxytocin receptor (OXTR) was assessed at the mRNA and protein levels by quantitative real-time PCR and immunoblotting respectively. Plasma and tissue activity of oxytocinase, the main enzyme involved in oxytocin degradation, were measured by fluorometric assay using an arylamide derivate as the substrate. Obese Zucker rats displayed a marked reduction in plasma oxytocin levels. Elevated liver and adipose tissue oxytocinase activity was noticed in obese Zucker rats. Hypothalamic oxytocin gene expression was not altered by the obese phenotype. OXTR mRNA and protein levels were upregulated in the adipose tissue of obese animals in contrast to the reduced OXTR protein levels in skeletal muscle. Our results show that obesity is associated with reduced plasma oxytocin due to increased peptide degradation by liver and adipose tissue rather than changes in hormone synthesis. This study highlights the importance of the oxytocin system in the pathogenesis of obesity and suggests oxytocinase inhibition as a candidate approach in the therapy of obesity.


Asunto(s)
Tejido Adiposo/metabolismo , Hígado/metabolismo , Obesidad Mórbida/metabolismo , Oxitocina/sangre , Animales , Cistinil Aminopeptidasa/sangre , Humanos , Masculino , Músculo Esquelético/metabolismo , Obesidad Mórbida/genética , Proteolisis , Ratas , Ratas Zucker , Receptores de Oxitocina/genética , Receptores de Oxitocina/metabolismo
20.
Behav Brain Res ; 213(2): 328-31, 2010 Dec 01.
Artículo en Inglés | MEDLINE | ID: mdl-20576539

RESUMEN

Individuals in the early stage of Parkinson's disease exhibit cognitive impairments as a result of hemisphere damage. The mesocortical dopamine system, particularly the medial prefrontal cortex (mPFC), is implicated in cognitive functions and is characterized by an asymmetric organization. Oxytocinase activity (OX) is also asymmetrically distributed in the mPFC of normal rats and is involved in cognitive functions. OX was measured in the left and right mPFC of rats with left or right hemi-parkinsonism, induced by intrastriatal injections of 6-hydroxydopamine, and compared with sham controls. These results demonstrated that the striking basal left predominance of OX observed in both the left and the right sham controls was radically disrupted in lesioned animals. The bilateral distribution in lesioned animals was altered differently depending on the injured hemisphere. These results may reflect changes in the enzyme substrates and consequently in the functions in which they are involved. These results may account, in part, for the cognitive abnormalities observed in hemi-parkinsonism.


Asunto(s)
Cistinil Aminopeptidasa/metabolismo , Modelos Animales de Enfermedad , Enfermedad de Parkinson/enzimología , Corteza Prefrontal/enzimología , Animales , Lateralidad Funcional , Masculino , Oxidopamina , Ratas , Ratas Wistar
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