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BACKGROUND: This study evaluated for the first time the potential of orange passion fruit as a base for alcoholic and acetic fermentations, with a view to assessing its profile of organic acids and polyphenols, in vitro digestion, and biological activities. RESULTS: In terms of aliphatic organic acids, malic acid was the majority in the wine (3.19 g L-1), while in the vinegar, it was acetic acid (46.84 g L-1). 3,4-Dihydroxybenzoic acid (3,4-DHB) was the major phenolic compound in the wine and vinegar samples (3443.93 and 2980.00 µg L-1, respectively). After the in vitro gastrointestinal simulation stage, the wine showed high bioaccessibility for the compounds sinipaldehyde (82.97%) and 2,4-dihydroxybenzoic acid (2,4-DHBA, 81.27%), while the vinegar exhibited high bioaccessibility for sinipaldehyde (89.39%). Through multivariate analysis, it was observed that 3,4-DHB was highly concentrated in the different digested fractions obtained from the wine. In contrast, in the vinegar, the stability of isorahmenetin and Quercetin 3-o-rhamnoside was observed during the in vitro digestion simulation. Lastly, the vinegar stood out for its inhibition rates of α-amylase (23.93%), α-glucoside (18.34%), and angiotensin-converting enzyme (10.92%). In addition, the vinegar had an inhibitory effect on the pathogenic microorganisms Salmonella enteritidis, Escherichia coli, and Listeria monocytogenes. CONCLUSION: Orange passion fruit has proved to be a promising raw material for the development of fermented beverages. Therefore, this study provides an unprecedented perspective on the use and valorization of orange passion fruit, contributing significantly to the advancement of knowledge about fermented products and the associated nutritional and functional possibilities. © 2024 Society of Chemical Industry.
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Ácido Acético , Digestión , Fermentación , Frutas , Passiflora , Fenoles , Vino , Passiflora/química , Passiflora/metabolismo , Frutas/química , Frutas/metabolismo , Ácido Acético/metabolismo , Ácido Acético/química , Ácido Acético/análisis , Fenoles/metabolismo , Fenoles/análisis , Fenoles/química , Vino/análisis , Humanos , Escherichia coli/efectos de los fármacos , Listeria monocytogenes/efectos de los fármacos , Malatos/análisis , Malatos/metabolismo , Extractos Vegetales/química , Extractos Vegetales/metabolismo , Extractos Vegetales/farmacología , Polifenoles/metabolismo , Polifenoles/análisis , Polifenoles/químicaRESUMEN
Many studies have reported the benefits of probiotic microorganisms and the production of angiotensin-converting enzyme (ACE) inhibitors. Determining the proteolytic and ACE inhibition capacities during whey fermentation was the goal of the study. Lacticaseibacillus rhamnosus GG, Streptococcus thermophilus SY-102, and both bacteria together were initially inoculated into whey, reaching an initial concentration of 108 CFU per milliliter in each fermentation system. Through the use of TNBS, SDS-PAGE, and SEC-HPLC methods, the proteolytic profile was examined. An in vitro investigation was performed to test the ACE inhibition capacity. With S. thermophilus, the logarithmic phase of microbial development was shorter than with L. rhamnosus (6 and 12 h, respectively). The logarithmic phase in the co-culture fermentation, however, was extended to 24 h. There were no significant differences in pH between the fermentations. However, the co-culture had a greater concentration of protein hydrolysis (453 ± 0.06 µg/mL), as indicated by the amount of free amino groups. Similarly, this fermentation produced more low molecular weight peptides. The higher inhibition activity, which increased at the conclusion of the fermentation with the co-culture and reached 53.42%, was influenced by the higher peptide synthesis. These findings highlighted the significance of creating useful co-culture products.
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Hypertension is the leading risk factor for cardiovascular diseases and is associated with intestinal dysbiosis with a decrease in beneficial microbiota. Probiotics can positively modulate the impaired microbiota and impart benefits to the cardiovascular system. Among them, the emended Lactobacillus has stood out as a microorganism capable of reducing blood pressure, being the target of several studies focused on managing hypertension. This review aimed to present the potential of Lactobacillus as an antihypertensive non-pharmacological strategy. We will address preclinical and clinical studies that support this proposal and the mechanisms of action by which these microorganisms reduce blood pressure or prevent its elevation.
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INTRODUCTION: Obesity-related metabolic diseases occur as a result of disruptions in white adipose tissue (WAT) plasticity, especially through visceral fat accumulation and adipocyte hypertrophy. This study aimed to evaluate the impact of renin-angiotensin system (RAS) and bradykinin receptors modulation by enalapril treatment and/or exercise training on WAT morphology and related deleterious outcomes. METHODS: Male C57BL/6 mice were fed either a standard chow or a high-fat (HF) diet for 16 weeks. At the 8th week, HF-fed animals were divided into sedentary (HF), enalapril treatment (HF-E), exercise training (HF-T), and enalapril treatment plus exercise training (HF-ET) groups. Following the experimental protocol, body mass gain, adiposity index, insulin resistance, visceral WAT morphometry, renin-angiotensin system, and bradykinin receptors were evaluated. RESULTS: The HF group displayed increased adiposity, larger visceral fat mass, and adipocyte hypertrophy, which was accompanied by insulin resistance, overactivation of Ang II/AT1R arm, and favoring of B1R in bradykinin receptors profile. All interventions ameliorated visceral adiposity and related outcomes by favoring the Ang 1-7/MasR arm and the B2R expression in B1R/B2R ratio. However, combined therapy additively reduced Ang II/Ang 1-7 ratio. CONCLUSION: Our results suggest that Ang 1-7/MasR arm and B2R activation might be relevant targets in the treatment of visceral obesity.
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Enalapril/farmacología , Condicionamiento Físico Animal/fisiología , Sistema Renina-Angiotensina/fisiología , Tejido Adiposo Blanco/metabolismo , Adiposidad/efectos de los fármacos , Adiposidad/fisiología , Animales , Dieta Alta en Grasa , Enalapril/metabolismo , Insulina/metabolismo , Resistencia a la Insulina/fisiología , Grasa Intraabdominal/efectos de los fármacos , Grasa Intraabdominal/fisiología , Masculino , Ratones , Ratones Endogámicos C57BL , Obesidad/tratamiento farmacológico , Obesidad/metabolismo , Obesidad Abdominal/metabolismo , Receptores de Bradiquinina/metabolismo , Sistema Renina-Angiotensina/efectos de los fármacosRESUMEN
The aim of this work was to determine the in vitro antihypertensive activities of lactobacillus (L. plantarum and L. helveticus) prepared amaranth protein hydrolysates, to determine the contribution of zinc, and to identify peptides. Depending on the bacteria species and the duration of the hydrolysis, up to 45.9% inhibition of angiotensin converting enzyme (ACE) was obtained. Size separation of the most active hydrolysates to yield < 1, <3-1, <3, <10-3 and < 10 kDa fractions enhanced ACE inhibition by 2-fold. A mixed mechanism of inhibition is proposed due to low correlation of ACE and zinc chelation. Thirty-six peptides were identified in the fractions using tandem mass spectrometry. A bioinformatic analysis showed the presence of encrypted fragments such as GVSEE or VNVDDPSK with known ACE-inhibitory properties. In conclusion, lactic acid bacteria proteases released peptides from amaranth proteins with ACE-inhibitory properties that were related to the presence of peptides with known or predicted ACE-inhibitor motifs.
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Amaranthus , Hidrolisados de Proteína , Angiotensinas , Hidrólisis , Lactobacillus , SemillasRESUMEN
Probiotic consumption promotes numerous health benefits. The aim of this study is 1) to evaluate the antihypertensive effect of kefir in a hypertension rat model caused by the administration of the nitric oxide synthesis inhibitor, L-NAME, and 2) to evaluate the acute angiotensin converting enzyme (ACE) inhibitory activity of the soluble nonbacterial fraction (SNBF) of kefir. To develop the first aim, male rats were separated into three groups: control group (C) treated with 0.3 mL/100 g of milk; L-NAME group (LN) received 10 mg/kg of said inhibitor; and Kefir group (K) treated with 0.3 mL/100 g of kefir plus L-NAME (10 mg/kg of said inhibitor). The treatments were given by oral gavage twice a day for four weeks. For the second aim"instead additionally, male rats received angiotensin I (in bolus) in three doses (Ang I: 0.03, 3 and 300 µg/kg) and were separated into two groups: a) received captopril (30 mg/kg i.v.) and b)received SNBF of kefir (5 mL/kg i.v.). Blood pressure were evaluated before and after Ang I. After treatment, hemodynamic parameters were evaluated, heart weight was recorded, and body weight gain was calculated. SNBF of kefir did not decrease the blood pressure for L-NAMEtreated animals, and no changes were observed in the cardiac parameters. However, the SNBF of kefir demonstrated acute inhibition of ACE in vivo similar to that of captopril. Thus, our results suggest that kefir may improve human cardiovascular systems by using mechanisms independent of nitric oxide syntheses. Additionally, the renin angiotensin system is probably the most important system involved in kefir effect regarding hypertension.
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Animales , Masculino , Ratas , Inhibidores de la Enzima Convertidora de Angiotensina , Kéfir/efectos adversos , Presión Sanguínea/genética , Probióticos/farmacología , Antihipertensivos/análisis , Óxido Nítrico/efectos adversosRESUMEN
Proteins and peptides are the most diverse biomolecules found in nature and make our interest due to their wide applications in food and pharmaceutical industry. Angiotensin Converting Enzyme (ACE) plays a major role in controlling blood pressure. The inhibition of ACE with peptides is a main target in the regulation of hypertension. The objective of the present study was to investigate the therapeutic potential of soy bean. This was accomplished by isolation of ACE inhibitory peptides using response surface methodology (RSM) and characterization of these bioactive peptides by mass spectrometry. 31 hydrolyzed fractions were isolated and evaluated for their ACE inhibition potential. Hydrolyzed fraction having highest ACE inhibitory activity was characterized by liquid chromatography-mass spectrometry (LC-MS) technique. RSM results showed maximum ACE inhibition potential (64%) by hydrolyzate was obtained at 45 ºC temperature, pH 8.0, E/S 0.2 in 2 hours hydrolysis time. Results of LC-MS analysis revealed Ser-Gly, Ser-Pro, Met-Ala, His-Ala, Lys-Pro, Phe-Thr, Met-Leu, Pro-Arg, Ala-Pro-Val, Pro-Ala-Leu, Val-Met-Gly, Pro-Leu-Val, Pro-Pro-Gln, His-Arg-Gly, Ser-Phe-Val-Leu, Ala-Val-His-Try, Arg-Thr-Val-Arg, His-His-Tyr-Leu-Val, Asp-Gly-Ala-Cys-Ser-Ala-Asn and MetVal-Thr-Gly-Pro-Gly-Cys-His bioactive peptides in hydrolyzed fraction of soy bean. Our data provide evidence that response surface methodology is a good approach for isolation of antihypertensive bioactive peptides with more potent activity as nutraceuticals or pharmaceuticals. Therefore soy bean can be use for industrial production of pharmaceutical grade natural medicines for handling high blood pressure.
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Péptidos/farmacología , Proteínas/farmacología , Proteínas de Soja/farmacología , Suplementos Dietéticos , Hidrolisados de Proteína/farmacología , Espectrometría de Masas , Cromatografía Liquida/métodos , Optimización de Procesos/clasificación , Concentración de Iones de Hidrógeno , Hipertensión/terapia , Antihipertensivos/análisisRESUMEN
The antihypertensive activity of the medicinal plant Hancornia speciosa has been previously demonstrated by us, being the activity ascribed to polyphenols and cyclitols like l-(+)-bornesitol. We herein evaluated the stability of the bioactive marker bornesitol submitted to forced degradation conditions. Bornesitol employed in the study was isolated from H. speciosa leaves. An UHPLC-ESI-MS/MS method was developed to investigate bornesitol stability based on MRM (Multiple Reaction Monitoring) acquisition mode and negative ionization mode, employing both specific (m/z 193â¯ââ¯161â¯Da) and confirmatory (m/z 193â¯ââ¯175â¯Da) transitions. A gradient elution of 0.1% formic acid in water and acetonitrile was performed on a HILIC column. The method was validated and showed adequate linearity (r2â¯>â¯0.99), selectivity, specificity, accuracy, and precision (RSDâ¯<â¯2.9%). The method was robust for deliberate variations on dessolvation temperature, but not for changes in the flow rate and dessolvation gas. The results from the stability studies allowed us to classify bornesitol as labile for acidic and alkaline hydrolysis, but as very stable for oxidative and neutral hydrolysis exposure. Bornesitol was categorized as practically stable under photolysis degradation, whereas a considerable reduction on its contents was induced by metal ions and thermolysis exposure. Degraded samples from neutral hydrolysis and thermolysis were assayed in vitro for ACE inhibition and showed a substantial decrease in biological activity as compared to intact bornesitol. myo-Inositol was identified as the major degradation products in both matrices. This is the first report on bornesitol stability under different stress conditions and the obtained data are relevant for the development and quality control of standardized products from H. speciosa leaves.
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Apocynaceae/química , Cromatografía Líquida de Alta Presión/métodos , Ciclitoles , Espectrometría de Masas/métodos , Peptidil-Dipeptidasa A/efectos de los fármacos , Inhibidores de la Enzima Convertidora de Angiotensina/análisis , Inhibidores de la Enzima Convertidora de Angiotensina/química , Inhibidores de la Enzima Convertidora de Angiotensina/farmacología , Biomarcadores/análisis , Biomarcadores/química , Ciclitoles/análisis , Ciclitoles/química , Ciclitoles/farmacología , Estabilidad de Medicamentos , Límite de Detección , Modelos Lineales , Peptidil-Dipeptidasa A/metabolismo , Extractos Vegetales/química , Reproducibilidad de los ResultadosRESUMEN
Cyanobacteria are able to produce a wide range of secondary metabolites, including toxins and protease inhibitors, with diverse biological activities. Microginins are small linear peptides biosynthesized by cyanobacteria species that act against proteases. The aim of this study was to isolate and identify microginins produced by the LTPNA08 strain of Microcystis aeruginosa, as well as to verify their potential to inhibit angiotensin-converting enzyme (ACE; EC. 3.4.15.1) using in vitro and in silico methods. The fractionation of cyanobacterial extracts was performed by liquid chromatography and the presence of microginins was monitored by both LC-MS and an ACE inhibition assay. Enzyme inhibition was assayed by ACE with hippuryl-histidyl-leucine as the substrate; monitoring of hippuric acid was performed by HPLC-DAD. Isolated microginins were confirmed by mass spectrometry and were used to carry out the enzymatic assay. Molecular docking was used to evaluate microginin 770 (MG 770) and captopril (positive control), in order to predict similar binding interactions and determine the inhibitory action of ACE. The enzyme assay confirmed that MG 770 can efficiently inhibit ACE, with an IC50 equivalent to other microginins. MG 770 presented with comparable interactions with ACE, having features in common with commercial inhibitors such as captopril and enalaprilate, which are frequently used in the treatment of hypertension in humans.
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Inhibidores de la Enzima Convertidora de Angiotensina/química , Antihipertensivos/química , Proteínas Bacterianas/química , Microcystis/química , Peptidil-Dipeptidasa A/química , Inhibidores de Proteasas/química , Inhibidores de la Enzima Convertidora de Angiotensina/aislamiento & purificación , Antihipertensivos/aislamiento & purificación , Proteínas Bacterianas/aislamiento & purificación , Sitios de Unión , Pruebas de Enzimas , Hipuratos/química , Humanos , Microcystis/metabolismo , Simulación del Acoplamiento Molecular , Oligopéptidos/química , Inhibidores de Proteasas/aislamiento & purificación , Unión Proteica , Conformación Proteica , Especificidad por SustratoRESUMEN
The aim of this work was to assess the effects of temperature (T), time (t) and pH treatments and an in vitro digestion on the stability of the angiotensin I-converting-enzyme-inhibitory activity (ACEIA) and antithrombotic activity (ATA; assessed as inhibition of platelet aggregation) of selected protein hydrolysates of amaranth named Alb1H103 and GloH88 and GluH24 with dipeptidyl peptidase IV inhibitory activity (DPPIVIA). Heat treatment (40-100 °C) for 1 h showed no significant differences among ACEIA, DPPIVIA and ATA of the heated hydrolysates at pH 4 and 7. There was no statistically significant loss of any bioactivity under heat treatment for 3 h at pH 4.0. Alb1H103 and GluH24 maintained the inhibitory activity of ACE and ATA at pH 7.0 for 3 h, whereas GloH88 maintained ACEIA and ATA for 2.0 h at pH 7.0. The pH effect on hydrolysates bioactivity was assessed in the range of 2.0-12.0. This was negligible on ACEIA, ATA and DPPIVIA. The in vitro digestion was performed using pepsin, trypsin (T) and α-chymotrypsin (C). A previous treatment of hydrolysates with pepsin improved the proteolytic activities of T and C. The hydrolysates kept at 100 °C for 1 h at pH 4.0, showed a significant increase in bioactivity. Conversely, a treatment at pH 7.0 showed no significant difference (p < 0.05) in the hydrolysates bioactivities after their digestion. Thus, biological activity of hydrolysates may be preserved or enhanced, depending on their processing conditions.
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Even though some research has been carried out on surfactant properties of amaranth protein hydrolysates, their bio-functionality has not been studied yet. In this work amaranth grain Alb 1 and Glob were hydrolyzed (Alb 1H, Glob H) and foams and emulsions at optimal conditions (t, E/S, pH5) were prepared in order to assess techno-functional properties such as foaming (F) and emulsifying (E) (capacity (C) and stability (S)). FC and EC were much better for Glob H than for Alb H. Angiotensin I-converting enzyme-inhibitory activity was higher for Alb 1H (roughly 50 %) than that of Glob H (roughly 30 %). Scavenging of radicals activity (DPPH· or ABTS· (+) ) of Alb 1H and Glob H, at 2 mg/mL, was similar (approx. 40 %), but lower than Alb 1 (approx. 70 %), which was the best antioxidant. The low reducing power showed that hydrolysates barely donate an electron or hydrogen. Chelating activity on Cu(2+) was lower than that exhibited by Fe(2+,) which was remarkable, approx. 80 % as long as DH% > 10 %, where hydrolysates displayed high solubility (Alb 1H = 85 %, Glob H = 70 %) because of occurrence of 1-10 kDa peptides. Amaranth foams and emulsions prepared with protein hydrolysates have a potential as a nutraceutical food.
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Angiotensin-converting enzyme inhibitors reduce blood pressure and attenuate cardiac and vascular remodeling in hypertension. However, the kinetics of remodeling after discontinuation of the long-term use of these drugs are unknown. Our objective was to investigate the temporal changes occurring in blood pressure and vascular structure of spontaneously hypertensive rats (SHR). Captopril treatment was started in the pre-hypertensive state. Rats (4 weeks) were assigned to three groups: SHR-Cap (N = 51) treated with captopril (1 g/L) in drinking water from the 4th to the 14th week; SHR-C (N = 48) untreated SHR; Wistar (N = 47) control rats. Subgroups of animals were studied at 2, 4, and 8 weeks after discontinuation of captopril. Direct blood pressure was recorded in freely moving animals after femoral artery catheterism. The animals were then killed to determine left ventricular hypertrophy (LVH) and the aorta fixed at the same pressure measured in vivo. Captopril prevented hypertension (105 ± 3 vs 136 ± 5 mmHg), LVH (2.17 ± 0.05 vs 2.97 ± 0.14 mg/g body weight) and the increase in cross-sectional area to luminal area ratio of the aorta (0.21 ± 0.01 vs 0.26 ± 0.02 ìm²) (SHR-Cap vs SHR-C). However, these parameters increased progressively after discontinuation of captopril (22nd week: 141 ± 2 mmHg, 2.50 ± 0.06 mg/g, 0.27 ± 0.02 ìm²). Prevention of the development of hypertension in SHR by using captopril during the prehypertensive period prevents the development of cardiac and vascular remodeling. Recovery of these processes follows the kinetic of hypertension development after discontinuation of captopril.