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1.
J Sci Food Agric ; 100(1): 315-324, 2020 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-31525262

RESUMO

BACKGROUND: In order to utilize tilapia skin gelatin hydrolysate protein, which is normally discarded as industrial waste in the process of fish manufacture, we study the in vivo and in vitro angiotensin-I-converting enzyme (ACE) inhibitory activity of the peptide Leu-Ser-Gly-Tyr-Gly-Pro (LSGYGP). The aim was to provide a pharmacological basis of the development of minimal side effects of ACE inhibitors by comparative analysis with captopril in molecular docking. RESULTS: This peptide from protein-rich wastes showed excellent ACE inhibitory activity (IC50  = 2.577 µmol L-1 ) and exhibited a mixed noncompetitive inhibitory pattern with Lineweaver-Burk plots. Furthermore, LSGYGP and captopril groups both showed significant decreases in blood pressure after 6 h and maintained good digestive stability over 4 h. Molecular bond interactions differentiate competitive captopril upon hydrogen bond interactions and Zn(II) interaction. The C-terminal Pro generates three interactions (hydrogen bonds, hydrophilic interactions and Van der Waals interactions) in the peptide and effectively interacts with the S1 and S2 pockets of ACE. CONCLUSION: LSGYGP, with an IC50 value of 2.577 µmol L-1 , has an antihypertensive effect in spontaneously hypertensive rats. Through comparison with captopril, this study revealed that LSGYGP may be a potential food-derived ACE inhibitory peptide and could act as a functional food ingredient to prevent hypertension. © 2019 Society of Chemical Industry.


Assuntos
Inibidores da Enzima Conversora de Angiotensina/química , Anti-Hipertensivos/química , Captopril/química , Hipertensão/tratamento farmacológico , Peptídeos/química , Sequência de Aminoácidos , Inibidores da Enzima Conversora de Angiotensina/administração & dosagem , Inibidores da Enzima Conversora de Angiotensina/metabolismo , Animais , Anti-Hipertensivos/administração & dosagem , Anti-Hipertensivos/metabolismo , Pressão Sanguínea/efeitos dos fármacos , Captopril/administração & dosagem , Ciclídeos , Digestão , Proteínas de Peixes/química , Trato Gastrointestinal/metabolismo , Humanos , Ligações de Hidrogênio , Interações Hidrofóbicas e Hidrofílicas , Hipertensão/metabolismo , Hipertensão/fisiopatologia , Cinética , Masculino , Simulação de Acoplamento Molecular , Peptídeos/metabolismo , Peptídeos/farmacologia , Peptidil Dipeptidase A/química , Peptidil Dipeptidase A/metabolismo , Hidrolisados de Proteína/química , Hidrolisados de Proteína/metabolismo , Ratos , Ratos Endogâmicos SHR
2.
Braz J Med Biol Res ; 52(11): e8772, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31664306

RESUMO

This study aimed to investigate the mechanism of fluorofenidone (AKF-PD) in treating renal interstitial fibrosis in rats with unilateral urinary obstruction (UUO). Thirty-two male Sprague-Dawley rats were randomly divided into sham, UUO, UUO + enalapril, and UUO + AKF-PD groups. All rats, except sham, underwent left urethral obstruction surgery to establish the animal model. Rats were sacrificed 14 days after surgery, and serum was collected for renal function examination. Kidneys were collected to observe pathological changes. Immunohistochemistry was performed to assess collagen I (Col I) protein expression, and terminal deoxynucleotidyl transferase-mediated nick end-labeling staining to observe the apoptosis of renal tubular epithelial cells. The expression of Fas-associated death domain (FADD), apoptotic protease activating factor-1 (Apaf-1), and C/EBP homologous protein (CHOP) proteins was evaluated by immunohistochemistry and western blot analysis. AKF-PD showed no significant effect on renal function in UUO rats. The pathological changes were alleviated significantly after enalapril or AKF-PD treatment, but with no significant differences between the two groups. Col I protein was overexpressed in the UUO group, which was inhibited by both enalapril and AKF-PD. The number of apoptotic renal tubular epithelial cells was much higher in the UUO group, and AKF-PD significantly inhibited epithelial cells apoptosis. The expression of FADD, Apaf-1, and CHOP proteins was significantly upregulated in the UUO group and downregulated by enalapril and AKF-PD. In conclusion, AKF-PD improved renal interstitial fibrosis by inhibiting apoptosis of renal tubular epithelial cells in rats with UUO.


Assuntos
Apoptose/efeitos dos fármacos , Células Epiteliais/efeitos dos fármacos , Nefropatias/patologia , Piridonas/farmacologia , Obstrução Ureteral/patologia , Inibidores da Enzima Conversora de Angiotensina/metabolismo , Inibidores da Enzima Conversora de Angiotensina/farmacologia , Animais , Fator Apoptótico 1 Ativador de Proteases/efeitos dos fármacos , Fator Apoptótico 1 Ativador de Proteases/metabolismo , Nitrogênio da Ureia Sanguínea , Colágeno Tipo I/efeitos dos fármacos , Colágeno Tipo I/metabolismo , Creatinina/sangue , Modelos Animais de Doenças , Enalapril/metabolismo , Enalapril/farmacologia , Proteína de Domínio de Morte Associada a Fas/efeitos dos fármacos , Proteína de Domínio de Morte Associada a Fas/metabolismo , Fibrose , Masculino , Piridonas/metabolismo , Distribuição Aleatória , Ratos Sprague-Dawley , Fator de Transcrição CHOP/efeitos dos fármacos , Fator de Transcrição CHOP/metabolismo
3.
Molecules ; 24(14)2019 Jul 12.
Artigo em Inglês | MEDLINE | ID: mdl-31336853

RESUMO

Angiotensin I-converting enzyme (ACE) is a paramount therapeutic target to treat hypertension. ACE inhibitory peptides derived from food protein sources are regarded as safer alternatives to synthetic antihypertensive drugs for treating hypertension. Recently, marine organisms have started being pursued as sources of potential ACE inhibitory peptides. Marine organisms such as fish, shellfish, seaweed, microalgae, molluscs, crustaceans, and cephalopods are rich sources of bioactive compounds because of their high-value metabolites with specific activities and promising health benefits. This review aims to summarize the studies on peptides from different marine organisms and focus on the potential ability of these peptides to inhibit ACE activity.


Assuntos
Inibidores da Enzima Conversora de Angiotensina/farmacologia , Organismos Aquáticos/metabolismo , Biossíntese Peptídica , Peptídeos/metabolismo , Peptídeos/farmacologia , Inibidores da Enzima Conversora de Angiotensina/química , Inibidores da Enzima Conversora de Angiotensina/metabolismo , Anti-Hipertensivos/metabolismo , Anti-Hipertensivos/farmacologia , Produtos Biológicos/metabolismo , Produtos Biológicos/farmacologia , Biomarcadores , Relação Dose-Resposta a Droga , Descoberta de Drogas , Avaliação Pré-Clínica de Medicamentos , Peptídeos/química
4.
Cells ; 8(6)2019 06 07.
Artigo em Inglês | MEDLINE | ID: mdl-31181665

RESUMO

Halophila stipulacea is a well-known invasive marine sea grass in the Mediterranean Sea. Having been introduced into the Mediterranean Sea via the Suez Channel, it is considered a Lessepsian migrant. Although, unlike other invasive marine seaweeds, it has not demonstrated serious negative impacts on indigenous species, it does have remarkable invasive properties. The present in-silico study reveals the biotechnological features of H. stipulacea by showing bioactive peptides from its rubisc/o protein. These are features such as antioxidant and hypolipideamic activities, dipeptidyl peptidase-IV and angiotensin converting enzyme inhibitions. The reported data open up new applications for such bioactive peptides in the field of pharmacy, medicine and also the food industry.


Assuntos
Hydrocharitaceae/metabolismo , Peptídeos/metabolismo , Inibidores da Enzima Conversora de Angiotensina/química , Inibidores da Enzima Conversora de Angiotensina/metabolismo , Antioxidantes/química , Antioxidantes/metabolismo , Dipeptidil Peptidase 4/química , Dipeptidil Peptidase 4/metabolismo , Hidrólise , Imunomodulação , Peptídeo Hidrolases/química , Peptídeo Hidrolases/metabolismo , Peptídeos/química , Proteínas de Plantas/química , Proteínas de Plantas/metabolismo , Ubiquitina/agonistas , Ubiquitina/metabolismo
5.
Nutrients ; 11(5)2019 Apr 30.
Artigo em Inglês | MEDLINE | ID: mdl-31052202

RESUMO

In this study we investigated the oligopeptide pattern in fermented cocoa beans and derived products after simulated gastrointestinal digestion. Peptides in digested cocoa samples were identified based on the mass fragmentation and on the software analysis of vicilin and 21 KDa cocoa seed protein sequences, the most abundant cocoa proteins. Quantification was carried out by liquid chromatography/electrospray ionisation mass spectrometry (LC/ESI-MS) using an internal standard. Sixty five peptides were identified in the digested samples, including three pyroglutamyl derivatives. The in vitro angiotensin-converting enzyme (ACE)-inhibitory activity of cocoa digests were tested, demonstrating a high inhibition activity, especially for digestates of cocoa beans. The peptides identified were screened for their potential ACE inhibitory activity through an in silico approach, and about 20 di-, three- and tetra-peptides actually present in our samples were predicted as active. Two of the potentially active peptides were chemically synthesized and then assessed for their inhibitory activity by using the ACE in vitro assay. These peptides demonstrated an ACE inhibitory activity, however, that was too weak to explain alone the high activity of cocoa digestates, suggesting a synergic effect of all cocoa peptides. As a whole, results showed that an average chocolate portion (30 g) ensures an amount of peptides after digestion that, assuming complete absorption, could reach almost a complete inhibition of ACE.


Assuntos
Inibidores da Enzima Conversora de Angiotensina/metabolismo , Cacau/química , Digestão , Oligopeptídeos/isolamento & purificação , Cromatografia Líquida de Alta Pressão , Humanos , Simulação de Acoplamento Molecular , Espectrometria de Massas por Ionização por Electrospray
6.
Med Chem ; 15(6): 574-587, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31084594

RESUMO

BACKGROUND: The Angiotensin-I converting enzyme (ACE) is one of the most important components of the renin-angiotensin-aldosterone system controlling blood pressure and renal functions. Inhibitors of ACE are first line therapeutics used in the treatment of hypertension and related cardiovascular diseases. Somatic ACE consists of two homologous catalytic domains, the C- and N-domains. Recent findings have shown that although both domains are highly homologous in structure, they may have different physiological functions. The C-domain is primarily involved in the control of blood pressure, in contrast to the N-domain that is engaged in the regulation of hematopoietic stem cell proliferation. The currently available ACE inhibitors have some adverse effects that can be attributed to the non-selective inhibition of both domains. In addition, specific Ndomain inhibitors have emerged as potential antifibrotic drugs. Therefore, ACE is still an important drug target for the development of novel domain-selective drugs not only for the cardiovascular system but also for other systems. OBJECTIVE: Detailed structural information about interactions in the protein-ligand complex is crucial for rational drug design. This review highlights the structural information available from crystallographic data which is essential for the development of domain selective inhibitors of ACE. METHODS: Over eighty crystal complexes of ACE are placed into the Protein Database. An overview of X-ray ACE complexes with various inhibitors in C- and N-domains and an analysis of their binding mode have given mechanistic explanation of the structural determinants of selective ligand binding. In addition, ACE domain selective inhibitors with dual modes of action in complexes with ACE are also discussed. CONCLUSION: Selectivity of ACE inhibitors for the N- and C-domain is controlled by subtle differences in the amino-acids forming the active site. Reported studies of crystal complexes of inhibitors in the C- and N-domains revealed that most selective inhibitors interact with non-conserved amino-acids between domains and have distinct interactions with the residues in the S2 and S2' subsites of the ACE catalytic site. Moreover, unusual binding of the second molecule of inhibitors in the binding cavity opens new possibilities of exploiting more distant regions of the catalytic center in structure-based design of novel drugs.


Assuntos
Inibidores da Enzima Conversora de Angiotensina/metabolismo , Peptidil Dipeptidase A/metabolismo , Inibidores da Enzima Conversora de Angiotensina/química , Animais , Domínio Catalítico , Cristalografia por Raios X , Desenho de Drogas , Humanos , Peptidil Dipeptidase A/química , Ligação Proteica , Domínios Proteicos , Especificidade por Substrato
7.
J Agric Food Chem ; 67(25): 7147-7156, 2019 Jun 26.
Artigo em Inglês | MEDLINE | ID: mdl-31140270

RESUMO

Egg proteins are recognized as excellent sources of bioactive peptides, such as angiotensin-converting enzyme inhibitory (ACEi) peptides. Oral administration of a thermolysin-digested egg white hydrolysate (T-EWH) caused a significant blood pressure reduction in spontaneously hypertensive rats; a further ACEi assay implied that its ACEi activity was enhanced after in vitro gastrointestinal (GI) digestion. These results indicated that T-EWH contained ACEi peptides resisting GI digestion and/or being further released during GI digestion. Therefore, the objective of this study was to identify these responsible ACEi peptides from T-EWH. The conventionally activity-guided fractionation was applied, coupled with a synchronized GI digestion throughout, during which both peptide yield and ACEi activity before and after the GI digestion were measured. Finally, six ACEi peptides (LAPYK, LKISQ, LKYAT, INKVVR, LFLIKH, and LGHWVY) with good GI resistance were identified with IC50 values <20 µM, especially LKYAT (0.09 µM). The structure-activity relationship of these peptides was discussed. The discovery of GI-resistant ACEi peptides could further support the application of egg white proteins as functional food ingredients.


Assuntos
Inibidores da Enzima Conversora de Angiotensina/química , Proteínas do Ovo/química , Trato Gastrointestinal/metabolismo , Hipertensão/metabolismo , Peptídeos/química , Inibidores da Enzima Conversora de Angiotensina/administração & dosagem , Inibidores da Enzima Conversora de Angiotensina/metabolismo , Animais , Pressão Sanguínea/efeitos dos fármacos , Galinhas , Digestão , Clara de Ovo/química , Hipertensão/tratamento farmacológico , Hipertensão/fisiopatologia , Peptídeos/administração & dosagem , Peptídeos/metabolismo , Peptidil Dipeptidase A/química , Hidrolisados de Proteína/química , Ratos , Ratos Endogâmicos SHR
8.
J Agric Food Chem ; 67(19): 5544-5551, 2019 May 15.
Artigo em Inglês | MEDLINE | ID: mdl-31007021

RESUMO

The purpose of the study was to investigate the transepithelial transport route of Arg-Leu-Ser-Phe-Asn-Pro (RLSFNP), a milk-derived angiotensin-converting enzyme (ACE) inhibitory peptide, and to encapsulate RLSFNP in a liposome to improve its intestinal bioavailability. The transport route was investigated using transport inhibitors in a human intestinal Caco-2 cell monolayer model. Sodium azide and wortmannin significantly reduced the permeability of RLSFNP ( P < 0.01), indicating that energy-dependent transcytosis is involved in the transport of RLSFNP across Caco-2 cells. The hexapeptide RLSFNP was then embedded in liposomes, and the RLSFNP liposome was characterized. Afterward, the cellular uptake and transepithelial transport ability of the RLSFNP liposome across Caco-2 cell monolayers was observed. The results demonstrated that the RLSFNP liposome was successfully prepared, having a significant sustained release and storage capability. The RLSFNP liposome can be absorbed by Caco-2 cells, with an increased intestinal absorption of RLSFNP compared to RLSFNP alone. The results showed a new way to improve RLSFNP intestinal bioavailability.


Assuntos
Células Epiteliais/metabolismo , Lipossomos/química , Peptídeos/metabolismo , Inibidores da Enzima Conversora de Angiotensina/química , Inibidores da Enzima Conversora de Angiotensina/metabolismo , Transporte Biológico , Células CACO-2 , Composição de Medicamentos , Humanos , Mucosa Intestinal/metabolismo , Lipossomos/metabolismo , Modelos Biológicos , Peptídeos/química , Peptidil Dipeptidase A/química , Peptidil Dipeptidase A/metabolismo
9.
J Food Sci ; 84(5): 1170-1179, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30997940

RESUMO

High blood pressure can lead to cardiovascular diseases. The objective of this work was to obtain protein hydrolysates with antihypertensive potential from chia oil industry meal byproduct. Chia seed protein isolates (CPIs) were obtained from chia seed meal byproduct. CPI was hydrolyzed using different proteases (alcalase, pepsin, trypsin, and α-chymotrypsin) and their biological potential was evaluated using in vitro and in silico approaches. Chia seed pepsin protein hydrolysate showed the highest angiotensin-converting enzyme inhibition potential IC50 of 0.128 mg/mL (P < 0.05) compared to the rest of hydrolysates. Peptide sequence LIVSPLAGRL presented the lowest predicted binding energy and highest inhibition potential (-9.5 kcal/mol) compared to other sequenced peptides and positive controls (captopril and lisinopril). Chia peptides showed potential to block angiotensin-converting enzyme by interacting with its catalytic site. Chia seed oil industry meal byproduct could be used as an inexpensive source of protein and bioactive peptides with antihypertensive potential. PRACTICAL APPLICATION: This research shows an upcycling alternative for chia oil industry byproduct. Chia meal is a rich source of protein and can be used to generate bioactive peptides with antihypertensive potential. Chia protein isolate was obtained from chia meal and hydrolyzed using different enzymes, pepsin showed the highest antihypertensive potential. Chia meal waste could be a low-cost source of protein and protein hydrolysates that could be used as a food ingredient with antihypertensive potential.


Assuntos
Inibidores da Enzima Conversora de Angiotensina , Anti-Hipertensivos , Pepsina A , Peptidil Dipeptidase A , Proteínas de Plantas , Salvia/química , Inibidores da Enzima Conversora de Angiotensina/química , Inibidores da Enzima Conversora de Angiotensina/metabolismo , Inibidores da Enzima Conversora de Angiotensina/farmacologia , Anti-Hipertensivos/química , Anti-Hipertensivos/metabolismo , Anti-Hipertensivos/farmacologia , Domínio Catalítico , Pepsina A/química , Pepsina A/metabolismo , Pepsina A/farmacologia , Peptidil Dipeptidase A/química , Peptidil Dipeptidase A/metabolismo , Proteínas de Plantas/química , Proteínas de Plantas/metabolismo , Proteínas de Plantas/farmacologia , Hidrolisados de Proteína/química , Hidrolisados de Proteína/metabolismo , Hidrolisados de Proteína/farmacologia
10.
Mar Drugs ; 17(3)2019 Mar 25.
Artigo em Inglês | MEDLINE | ID: mdl-30934583

RESUMO

Plastid proteins are one of the main components in red algae. In order to clarify the angiotensin I converting enzyme (ACE) inhibitory peptides from red alga Palmaria sp. (Japan), we determined the plastid genome sequence. The genome possesses 205 protein coding genes, which were classified as genetic systems, ribosomal proteins, photosystems, adenosine triphosphate (ATP) synthesis, metabolism, transport, or unknown. After comparing ACE inhibitory peptides between protein sequences and a database, photosystems (177 ACE inhibitory peptides) were found to be the major source of ACE inhibitory peptides (total of 751). Photosystems consist of phycobilisomes, photosystem I, photosystem II, cytochrome complex, and a redox system. Among them, photosystem I (53) and II (51) were the major source of ACE inhibitory peptides. We found that the amino acid sequence of apcE (14) in phycobilisomes, psaA (18) and psaB (13) in photosystem I, and psbB (11) and psbC (10) in photosystem II covered a majority of bioactive peptide sequences. These results are useful for evaluating the bioactive peptides from red algae.


Assuntos
Rodófitas/genética , Sequência de Aminoácidos , Inibidores da Enzima Conversora de Angiotensina/metabolismo , Inibidores da Enzima Conversora de Angiotensina/farmacologia , Simulação por Computador , Genomas de Plastídeos , Fases de Leitura Aberta , Peptídeos/metabolismo , Peptídeos/farmacologia , Rodófitas/metabolismo , Sequenciamento Completo do Genoma
11.
Mar Drugs ; 17(4)2019 Mar 30.
Artigo em Inglês | MEDLINE | ID: mdl-30935056

RESUMO

A protein extract was generated from the macroalga Ulva lactuca, which was subsequently hydrolysed using the food-grade enzyme papain and angiotensin-converting Enzyme I and renin inhibitory peptides identified using a combination of enrichment strategies employing molecular weight cutoff filtration and mass spectrometry analysis. The generated hydrolysates with the most promising in vitro activity were further purified using preparative RP-HPLC and characterised. The 1 kDa hydrolysate (1 kDa-UFH), purified and collected by preparative RP-HPLC at minutes 41‒44 (Fr41‒44), displayed statistically higher ACE-I inhibitory activities ranging from 96.91% to 98.06%. A total of 48 novel peptides were identified from these four fractions by LC-MS/MS. A simulated gastrointestinal digestion of the identified peptide sequences was carried out using in silico enzyme cleavage simulation tools, resulting in 86 peptide sequences that were further assessed for their potential activity, toxicity and allergenicity using multiple predictive approaches. All the peptides obtained in this study were predicted to be non-toxic. However, 28 out of the 86 novel peptides released after the in silico gastrointestinal digestion were identified as potential allergens. The potential allergenicity of these peptides should be further explored to comply with the current labelling regulations in formulated food products containing U. lactuca protein hydrolysates.


Assuntos
Inibidores da Enzima Conversora de Angiotensina/metabolismo , Inibidores da Enzima Conversora de Angiotensina/farmacologia , Oligopeptídeos/metabolismo , Oligopeptídeos/farmacologia , Hidrolisados de Proteína/farmacologia , Ulva/metabolismo , Alérgenos/farmacologia , Sequência de Aminoácidos , Inibidores da Enzima Conversora de Angiotensina/química , Inibidores da Enzima Conversora de Angiotensina/isolamento & purificação , Anti-Hipertensivos/farmacologia , Simulação por Computador , Humanos , Hidrólise , Oligopeptídeos/química , Oligopeptídeos/isolamento & purificação , Hidrolisados de Proteína/química , Hidrolisados de Proteína/isolamento & purificação , Alga Marinha/química , Ulva/química , Ulva/citologia
12.
Nefrología (Madrid) ; 39(2): 184-191, mar.-abr. 2019. tab, graf
Artigo em Espanhol | IBECS | ID: ibc-181326

RESUMO

Antecedentes y objetivo: Las enzimas metaloproteinasas de matriz (MMP) están involucradas en el remodelado tisular deletéreo asociado al daño de órganos diana de la enfermedad renal. El objetivo de este estudio fue explorar la asociación entre la caída de la función renal y la actividad sistémica de la metaloproteinasa inflamatoria MMP-9 en el paciente hipertenso con enfermedad renal crónica (ERC) leve-moderada. Material y métodos: Se analizaron los niveles plasmáticos de MMP-9 activa, MMP-9 total, su inhibidor tisular (TIMP-1), el cociente MMP-9/TIMP-1 y la interacción entre MMP-9 y TIMP-1 en 37 pacientes hipertensos distribuidos según su tasa de filtración glomerular estimada (TFGe) en 3 grupos: > 90, 90-60 y 60-30 mL/min/1,73m2. Resultados: La MMP-9 total no fue diferente con respecto a la disminución en la TFGe. TIMP-1 estaba significativamente incrementado en los pacientes hipertensos con TFGe entre 60-30 mL/min/1,73 m2 (p < 0,01 versus > 90 mL/min/1,73 m2). Estos resultados fueron apoyados por la disminución significativa de la interacción MMP-9-TIMP-1 observada en los pacientes con TFGe entre 60-30 mL/min/1,73 m2 (p < 0,01 versus > 90 mL/min/1,73 m2). A pesar de la elevación sistémica de TIMP-1 encontramos un incremento significativo de MMP-9 activa en los pacientes hipertensos con TFGe entre 60-30 mL/min/1,73m2 (p < 0,05 y p < 0,01 versus > 90 y 90-60 mL/min/1,73 m2, respectivamente). Los niveles de TIMP-1, MMP-9 activa e interacción proteica MMP-9-TIMP-1 correlacionaron significativamente con el deterioro de la función renal, lo cual no se observó para la MMP-9 total. Conclusiones: La progresión de la ERC, incluso en estadios donde la caída de la función renal es aún moderada, se asocia con un aumento específico de la actividad MMP-9, lo cual podría considerarse como una potencial diana terapéutica


Background and objective: Matrix metalloproteinases (MMPs) are involved in deleterious tissue remodeling associated with target organ damage in renal disease. The aim of this study was to study the association between renal dysfunction and activity of the inflammatory metalloproteinase MMP-9 in hypertensive patients with mild-moderate chronic kidney disease (CKD). Material and methods: Plasmatic active MMP-9, total MMP-9, tissue inhibitor of MMP-9 (TIMP-1), MMP-9/TIMP-1 ratio and MMP-9-TIMP-1 interaction were analyzed in 37 hypertensive patients distributed by estimated glomerular filtration rate (eGFR) in 3 groups:> 90, 90-60 y 60-30 mL/min/1.73 m2. Results: Total MMP-9 was not different as eGFR declines. TIMP-1 was significantly increased in hypertensive patients with eGFR 60-30 mL/min/1.73 m2 (P < .01 versus > 90 mL/min/1.73 m2). This relates to the significant decrease in the interaction between MMP-9-TIMP-1 observed in patients with eGFR 60-30mL/min/1.73 m2 (P < .01 versus > 90 mL/min/1.73 m2). Despite the systemic elevation of TIMP-1, active MMP-9 was significantly increased in hypertensive patients with eGFR 60-30 mL/min/1.73 m2 (P<.05 and P < 0.01 versus > 90 and 90-60 mL/min/1.73 m2, respectively). TIMP-1, active MMP-9 and MMP-9-TIMP-1 interaction significantly correlate with the decline in renal function, which was not observed with total MMP-9. Conclusions: The progression of CKD, even in stages where the decline of renal function is still moderate, is associated with an increase in MMP-9 activity, which could be considered as a potential therapeutic target


Assuntos
Humanos , Masculino , Pessoa de Meia-Idade , Idoso , Metaloproteinase 9 da Matriz/metabolismo , Insuficiência Renal Crônica/enzimologia , Hipertensão Essencial/complicações , Taxa de Filtração Glomerular , Análise de Variância , Inibidores da Enzima Conversora de Angiotensina/metabolismo
13.
J Biotechnol ; 294: 67-72, 2019 Mar 20.
Artigo em Inglês | MEDLINE | ID: mdl-30772329

RESUMO

This study focused on a culture system of aerial microalgae with the decomposition of casein protein for obtaining bioactive compounds such as peptides with inhibitory activity against angiotensin-converting enzyme (ACE). The aerial microalga Vischeria helvetica exhibited growth in Bold's basal medium supplemented with casein protein as nitrogen source. The algal cells secreted protease and amino oxidase into the medium, and ammonium ions as a nitrogen source was produced by the conjugated-enzyme reaction. Furthermore, a bioactive peptide with ACE-inhibitory activity was efficiently produced from casein protein by the proteases secreted under light conditions. The results presented will facilitate the development of production systems for useful materials from photosynthetic microorganisms and casein protein in a culture medium.


Assuntos
Inibidores da Enzima Conversora de Angiotensina/metabolismo , Caseínas/metabolismo , L-Aminoácido Oxidase/metabolismo , Microalgas/metabolismo , Oligopeptídeos/metabolismo , Peptídeo Hidrolases/metabolismo , Estramenópilas/metabolismo , Proteínas de Algas/metabolismo , Hidrólise , Peptidil Dipeptidase A/metabolismo
14.
Recent Pat Biotechnol ; 13(3): 239-248, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30747089

RESUMO

BACKGROUND: Hypertension is the chronic medical condition and it affected billions of people worldwide. Natural medicines are the main alternatives to treatment for a majority of people suffering from hypertension. Niazicin-A, Niazimin-A, and Niaziminin-B compounds from Moringa oleifera ethanolic leave extract were reported to have potent antihypertensive activity. OBJECTIVE: These compounds were targeted with Angiotensin-converting enzyme [ACE] which is one of the main regulatory enzymes of the renin-angiotensin system. METHODS: Protein-ligand docking of these compounds with [ACE] [both domain N and C] was conceded out through Autodock vina and visualization was done by chimera. Pharmacokinetics study of these compounds was predicted by ADME-Toxicity Prediction. RESULTS: Niazicin-A, Niazimin-A, and Niaziminin-B showed high binding affinity with ACE and partially blocked the active sites of the enzyme. Niazicin-A, Niazimin-A and Niaziminin-B showed the estimated free binding energy of -7.6kcal/mol kcal/mol, -8.8kcal/mol and -8.0kcal/mol respectively with C-domain of ACE and -7.9kcal/mol, -8.5kcal/mol and -7.7kcal/mol respectively with N-domain of ACE. The compounds showed better binding energy with angiotensinconverting enzyme in comparison to Captopril -5.5kcal/mol and -5.6kcal/mol and Enalapril [standard] -8.4kcal/mol and -7.5kcal/mol with C and N domain, respectively. CONCLUSION: Computationally, the selected bioactive molecules have shown better binding energy to known standard drugs which have been already known for inhibition of ACE and can further act as a pharmacophore for in vitro and in vivo studies in the development of alternative medicine.


Assuntos
Inibidores da Enzima Conversora de Angiotensina/química , Anti-Hipertensivos/química , Moringa oleifera/química , Peptidil Dipeptidase A/química , Tiocarbamatos/química , Inibidores da Enzima Conversora de Angiotensina/isolamento & purificação , Inibidores da Enzima Conversora de Angiotensina/metabolismo , Anti-Hipertensivos/isolamento & purificação , Anti-Hipertensivos/metabolismo , Captopril/química , Captopril/metabolismo , Domínio Catalítico , Enalapril/química , Enalapril/metabolismo , Expressão Gênica , Humanos , Hipertensão/tratamento farmacológico , Hipertensão/enzimologia , Cinética , Simulação de Acoplamento Molecular , Patentes como Assunto , Peptidil Dipeptidase A/genética , Peptidil Dipeptidase A/metabolismo , Extratos Vegetais/química , Folhas de Planta/química , Ligação Proteica , Conformação Proteica em alfa-Hélice , Conformação Proteica em Folha beta , Domínios e Motivos de Interação entre Proteínas , Especificidade por Substrato , Termodinâmica , Tiocarbamatos/isolamento & purificação , Tiocarbamatos/metabolismo
15.
Artigo em Inglês | MEDLINE | ID: mdl-30776402

RESUMO

Individuals with fear-associated conditions such as panic disorder (PD) and posttraumatic stress disorder (PTSD) display increased emotional responses to interoceptive triggers, such as CO2 inhalation, that signal a threat to physiological homeostasis. Currently, effector systems and mechanisms underlying homeostatic modulation of fear memory are not well understood. In this regard, the renin angiotensin system (RAS), particularly the angiotensin receptor type 1 (AT1R), a primary homeostatic regulatory target, has gained attention. RAS polymorphisms have been reported in PD and PTSD, and recent studies report AT1R-mediated modulation of fear extinction. However, contribution of AT1Rs in fear evoked by the interoceptive threat of CO2 has not been investigated. Using pharmacological, behavioral, and AT1R/ACE gene transcription analyses, we assessed central AT1R recruitment in CO2-associated fear. CO2 inhalation led to significant AT1R and ACE mRNA upregulation in homeostatic regulatory regions, subfornical organ (SFO) and paraventricular nucleus (PVN), in a temporal manner. Intracerebroventricular infusion of selective AT1R antagonist, losartan, significantly attenuated freezing during CO2 inhalation, and during re-exposure to CO2 context, suggestive of AT1R modulation of contextual fear. Regional Fos mapping in losartan-treated mice post-behavior revealed significantly attenuated labeling in areas regulating defensive behavior, contextual fear, and threat responding; such as, the bed nucleus of stria terminalis, dorsal periaqueductal gray, hypothalamic nuclei, hippocampus, and prefrontal areas such as the prelimbic, infralimbic, and anterior cingulate cortices. Sub-regions of the amygdala did not show CO2-associated AT1R regulation or altered Fos labeling. Collectively, our data suggests central AT1R recruitment in modulation of fear behaviors associated with CO2 inhalation via engagement of neurocircuits regulating homeostasis and defensive behaviors. Our data provides mechanistic insights into the interoceptive regulation of fear, relevant to fear related disorders such as PD and PTSD.


Assuntos
Dióxido de Carbono/metabolismo , Medo/fisiologia , Vias Neurais/fisiologia , Receptor Tipo 1 de Angiotensina/fisiologia , Bloqueadores do Receptor Tipo 1 de Angiotensina II/farmacologia , Inibidores da Enzima Conversora de Angiotensina/metabolismo , Animais , Encéfalo/fisiologia , Reação de Congelamento Cataléptica/efeitos dos fármacos , Homeostase/fisiologia , Infusões Intraventriculares , Losartan/farmacologia , Masculino , Camundongos , Núcleo Hipotalâmico Paraventricular/metabolismo , Receptor Tipo 1 de Angiotensina/metabolismo , Órgão Subfornical/metabolismo , Regulação para Cima
16.
Peptides ; 112: 34-42, 2019 02.
Artigo em Inglês | MEDLINE | ID: mdl-30481537

RESUMO

Hypertension is the fundamental cause of cardiovascular and cerebrovascular disorders. Several natural and synthetic peptides are being used as antihypertensive agents, which target angiotensin converting enzyme (ACE), the master regulator of angiotensin (Ang) II production. In this study, we have evaluated ACE-inhibitory potential of the tripeptide l-Phenylalanyl-d-Histidyl-l-Leucine (l-Phe-d-His-l-Leu) in vitro and its antihypertensive effect in rat model of dexamethasone-induced hypertension. l-Phe-d-His-l-Leu was custom-designed by changing the configuration of penultimate amino acid residue (histidine) from C-terminal of Ang I, the site at which ACE acts upon and generates Ang II. l-Phe-d-His-l-Leu effectively inhibited ACE activity in a dose-dependent and competitive manner with an IC50 of 53.32 ± 0.13 nmol/L. Both fluorescence spectra and circular dichroism data revealed the direct interaction between l-Phe-d-His-l-Leu and ACE. In addition, molecular docking studies revealed the strong interaction of l-Phe-d-His-l-Leu with the critical active site amino acid residues of ACE. Further, the administration of l-Phe-d-His-l-Leu resulted in decrease in blood pressure (142 ± 3 mmHg) compared to dexamethasone alone group (167 ± 2 mmHg). Besides, l-Phe-d-His-l-Leu decreased the levels of circulating Ang II, and reduced fibrosis in heart and kidney, as evidenced by decreases in collagen deposition. Thus, the strategy of incorporation of d-amino acids in ACE-inhibitory peptides could be valuable in the development of antihypertensive drugs.


Assuntos
Inibidores da Enzima Conversora de Angiotensina/farmacologia , Modelos Animais de Doenças , Hipertensão/tratamento farmacológico , Simulação de Acoplamento Molecular , Oligopeptídeos/farmacologia , Peptidil Dipeptidase A/efeitos dos fármacos , Angiotensina II/sangue , Angiotensina II/metabolismo , Inibidores da Enzima Conversora de Angiotensina/metabolismo , Inibidores da Enzima Conversora de Angiotensina/uso terapêutico , Animais , Domínio Catalítico , Dexametasona/toxicidade , Hipertensão/induzido quimicamente , Hipertensão/enzimologia , Hipertensão/fisiopatologia , Cinética , Masculino , Oligopeptídeos/metabolismo , Oligopeptídeos/uso terapêutico , Peptidil Dipeptidase A/química , Peptidil Dipeptidase A/metabolismo , Ratos , Ratos Wistar , Sistema Renina-Angiotensina
17.
Artigo em Inglês | MEDLINE | ID: mdl-30508739

RESUMO

Angiotensin converting enzyme (ACE, peptidyldipeptidase A, EC 3.4.15.1) plays an important role in the regulation of blood pressure. In this study, ACE was purified from human plasma by affinity chromatography in single step. The enzyme purified in 5367-fold from human plasma and specific activity was found to be 1208 EU/mg protein. The purity and molecular weight of ACE were determined by SDS-PAGE, which indicated two bands at around 60 kDa and 70 kDa on the gel. Effect of oxidized glutathione (GSSG) peptide and reduced glutathione (GSH) peptide on purified ACE activity were also investigated in which lisinopril was used as reference inhibitor. GSSG showed activation effect on ACE activity whereas GSH provided inhibition effect. In the lights of activity (%) versus activator graph for GSSG and activity (%) versus inhibitor graphs for GSH and lisinopril; IC50 values for GSH and lisinopril were determined to be 16.2 µM and 0.781 nM, respectively. Type of inhibition for GSH and lisinopril from graph Lineweaver-Burk was found to be reversible non-competitive inhibition and Ki constants for GSH and lisinopril were calculated as 11.7 µM and 0.662 nM, respectively.


Assuntos
Inibidores da Enzima Conversora de Angiotensina/metabolismo , Dissulfeto de Glutationa/metabolismo , Glutationa/metabolismo , Fragmentos de Peptídeos/metabolismo , Peptidil Dipeptidase A , Inibidores da Enzima Conversora de Angiotensina/química , Cromatografia de Afinidade , Glutationa/química , Dissulfeto de Glutationa/química , Humanos , Modelos Lineares , Fragmentos de Peptídeos/química , Peptidil Dipeptidase A/sangue , Peptidil Dipeptidase A/isolamento & purificação , Peptidil Dipeptidase A/metabolismo
18.
Biol Pharm Bull ; 42(1): 116-122, 2019 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-30369530

RESUMO

The emulsion prepared with ß-cyclodextrin as an emulsifier (ßCDE) is considered to be a Pickering emulsion. We examined the characteristics of ßCDEs using captopril (CP) as a model drug, and studied the in vitro skin permeation of CP from ßCDEs through hairless mouse skin. The stability of ßCDE was increased with increasing ßCD concentration and conversely decreased with increasing CP concentration. The yield stress value from the rheological measurement results was suggested to be one of the factors determining the stability of the ßCDE, and ßCDEs with higher yield stress values were more stable. We found that the skin permeability of CP could be improved by using ßCDE with isopropyl myristate as the oil phase and that the flux of CP depended on the free CP concentration in the water phase of ßCDE.


Assuntos
Ciclodextrinas/administração & dosagem , Ciclodextrinas/metabolismo , Sistemas de Liberação de Medicamentos/métodos , Emulsificantes/administração & dosagem , Emulsificantes/metabolismo , Absorção Cutânea/efeitos dos fármacos , Administração Cutânea , Inibidores da Enzima Conversora de Angiotensina/administração & dosagem , Inibidores da Enzima Conversora de Angiotensina/metabolismo , Animais , Captopril/administração & dosagem , Captopril/metabolismo , Camundongos , Camundongos Pelados , Técnicas de Cultura de Órgãos , Absorção Cutânea/fisiologia
19.
Probiotics Antimicrob Proteins ; 11(1): 273-282, 2019 03.
Artigo em Inglês | MEDLINE | ID: mdl-29209903

RESUMO

The potential for reusability of whey is of concern due to its substantial nutritional value and, second, in view of the need to reduce environmental impact. The use of goat whey powder in the production of a fermented dairy beverage using a Lactobacillus casei culture as adjunct shows a good prospect of a food product with hypotensive activity. This study investigated the microbial viability, proteolysis and angiotensin-converting enzyme (ACE) inhibitory activity of a fermented dairy beverage produced with goat whey powder and a probiotic culture of L. casei BGP93 co-cultured with Streptococcus thermophilus TA-40. The probiotic beverage exhibited no significant difference from the control (absence of L. casei) with regard to titratable acidity, S. thermophilus viability and proteolysis degree during 21 days of storage (P > 0.05). During this period, the beverage maintained L. casei at appropriate levels (> 7 log cfu ml-1), thereby qualifying as a potential probiotic product. Although both control and probiotic beverages exhibited ACE inhibitory activity, as a result of proteolysis of whey proteins during fermentation, significant increased ACE inhibitory activity was found for the beverage with added probiotic L. casei (P < 0.05). The probiotic beverage has potency as a functional food candidate to be included in a dietary strategy aiming at prevention and control of hypertension.


Assuntos
Inibidores da Enzima Conversora de Angiotensina/metabolismo , Produtos Fermentados do Leite/microbiologia , Lactobacillus casei/metabolismo , Streptococcus thermophilus/metabolismo , Proteínas do Soro do Leite/metabolismo , Inibidores da Enzima Conversora de Angiotensina/química , Animais , Bebidas/análise , Bebidas/microbiologia , Técnicas de Cocultura , Produtos Fermentados do Leite/análise , Fermentação , Cabras , Lactobacillus casei/crescimento & desenvolvimento , Leite/química , Leite/microbiologia , Peptidil Dipeptidase A/química , Proteólise , Streptococcus thermophilus/crescimento & desenvolvimento , Proteínas do Soro do Leite/química
20.
J Dairy Sci ; 102(2): 961-975, 2019 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-30594363

RESUMO

Lactobacillus helveticus LB 10 proteinases immobilized with sodium alginate were used to hydrolyze whey protein to produce angiotensin-I-converting enzyme (ACE)-inhibitory peptides. The generated hydrolysates were tested for ACE-inhibitory activity and for their ability to be transported across Caco-2 cell monolayers. Using a response surface method, we determined that a proteinase concentration of 7.55 mg/mL, sodium alginate concentration of 2.03 g/100 mL, and glutaraldehyde concentration of 0.39% were found to be the optimal immobilization conditions. Compared with free proteinase, the immobilized proteinase had significantly higher pH, thermal and storage stability, and reusability. Whey protein hydrolysates were fractionated by gel filtration chromatography and ACE-inhibitory peptide mixtures were transported across Caco-2 cell monolayers in a human intestinal-absorption model. The di- and tripeptides KA, EN, DIS, EVD, LF, AIV, and VFK (half-maximal inhibitory concentrations (mean ± standard deviation) of 1.24 ± 0.01, 1.43 ± 0.04, 1.59 ± 0.27, 1.32 ± 0.05, 1.60 ± 0.39, 2.66 ± 0.02, and 1.76 ± 0.09 mmol/L, respectively) were detected on the basolateral side of the Caco-2 cell monolayer using ultra-performance liquid chromatography-tandem mass spectrometry. These results highlight that ACE-inhibitory peptides are present on the basolateral side of the Caco-2 cell model after transportation of whey protein hydrolysate across the Caco-2 cell membrane.


Assuntos
Inibidores da Enzima Conversora de Angiotensina/metabolismo , Enzimas Imobilizadas/metabolismo , Lactobacillus helveticus/enzimologia , Peptídeo Hidrolases/metabolismo , Proteínas do Soro do Leite/metabolismo , Animais , Transporte Biológico , Células CACO-2 , Membrana Celular/metabolismo , Humanos , Hidrólise , Peptidil Dipeptidase A/metabolismo , Proteólise
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