RESUMEN
Consideradas o maior problema global de saúde, as doenças crônicas não transmissíveis (DCNT) são a causa de grande parte das morbimortalidades no Brasil atualmente e possuem fatores de risco comuns, o que possibilita uma abordagem preventiva semelhante. Sabendo que a escola é um local fundamental para promoção em saúde, este artigo tem como objetivo relatar a experiência de um projeto de extensão universitária que foi realizado com alunos do ensino fundamental, em duas escolas públicas do município de Juiz de Fora MG, nos anos de 2018 e 2019. Foram realizadas palestras expositivas e dinâmicas sobre os sistemas do corpo humano e a sua relação com as DCNT, tendo sido aplicado um questionário, no início e fim da intervenção, a fim de avaliar o desempenho e compreensão dos alunos. Foram feitas estatísticas descritivas com valores absolutos, mínimos e máximos, médias e porcentagens. Após isso, foi possível constatar que os alunos obtiveram uma melhora significativa do desempenho não apenas no questionário, mas também no âmbito social, pois foi perceptível a troca de saberes entre alunos e acadêmicos, o que proporcionou um ambiente dinâmico e favorável para o aprendizado.
Considered the biggest global health problem, non-communicable chronic diseases (NCDs) are the cause of most morbidity and mortality in Brazil today and have common risk factors, which allows for a similar preventive approach. Knowing that school is a fundamental place for health promotion, this article aims to report the experience of a university extension project, carried out with elementary school students,in two public schools in the city of Juiz de Fora -MG, in the years from 2018 and 2019. Expository and dynamic lectures were held on human body systems and their relationship with CNCDs, with a questionnaire being applied at the beginning and end of the intervention, in order to assess the performance and understanding of students. Descriptive statistics were performed with absolute, minimum and maximum values, means and percentages. After that, it was possible to verify that the students obtained a significant performance improvement not only in the questionnaire, but also in the social sphere, as the exchange of knowledge between students and academics was noticeable, which provided a dynamic and favorable environment for learning.
Asunto(s)
Educación en Salud , Adolescente , Enfermedades no TransmisiblesRESUMEN
OBJECTIVES: This study aimed to evaluate the potential of aqueous extract from Mitracarpus frigidus aerial parts (MFAq) in the treatment of inflammation and oxidative stress, as well as to characterize its chemical constituents. METHODS: Total phenolic and flavonoid contents were determined, and phytoconstituents were detected by ultra-fast liquid chromatography/quadrupole-time-of-flight tandem mass spectrometry (UFLC-QTOF-MS). The antioxidant activity was evaluated by DPPH, TAC and ß-carotene/linoleic acid assays. In-vitro anti-inflammatory activity, cell viability and cell cycle were performed in J774A.1 cell line. In-vivo anti-inflammatory activity was investigated by two ear oedema assays (croton oil and phenol). KEY FINDINGS: Chlorogenic acid, clarinoside, quercetin-hexosylpentoside, rutin, kaempferol-3-O-rutinoside, kaempferol-rhamnosylhexoside, quercetin-pentosylrhamnosylhexoside, harounoside, 2-azaanthraquinone and sucrose were identified by UFLC-QTOF-MS. MFAq showed antioxidant activity, which was positively correlated to the content of phenolic compounds. MFAq significantly inhibited the production of nitric oxide, did not decrease viability in MTT assay (all concentrations) and showed no changes in membrane permeability and cell cycle of J774A.1 cell line. Furthermore, MFAq showed a reduction in ear oedema in all tested doses. CONCLUSION: MFAq was effective in some antioxidant and inflammatory parameters, in the experimental conditions that were used in the study. This is the first report of chemical composition and bioactivities from this extract.
Asunto(s)
Rubiaceae , Antiinflamatorios/uso terapéutico , Antioxidantes/química , Edema/inducido químicamente , Edema/tratamiento farmacológico , Humanos , Fenoles/farmacología , Extractos Vegetales/química , Quercetina , Rubiaceae/químicaRESUMEN
Centrosema coriaceum Benth belongs to Fabaceae family and have few studies of biological activity and chemical composition. Thus, the aims of this work were to determine chemical profile of the ethanolic extract of C. coriaceum leaves (CCE) by UFLC-QTOF-MS and to evaluate its in vitro biological potential. CCE showed MIC value of 1000 µg/mL against Candida glabrata (fungistatic effect) and high affinity in cell envelope by increasing cell permeability in nucleotide leakage, sorbitol and ergosterol assays. CCE showed antioxidant activity in all assays performed. For the anti-inflammatory and cytotoxicity activities, CCE, at all tested concentrations, significantly inhibited the production of nitric oxide and did not decrease J774A.1 cell viability below 70%. Finally, rutin, kaempferol-3O-rutinoside, caffeic acid, and sucrose were identified in CCE by UFLC-QTOF-MS. These results suggest, for the first time, that C. coriaceum has interesting antifungal, antioxidant, and anti-inflammatory activities.
Asunto(s)
Fabaceae , Antiinflamatorios/farmacología , Antioxidantes/farmacología , Extractos Vegetales/farmacología , Hojas de la PlantaRESUMEN
AIMS: We aimed to determine whether resistance training (RT) regulates renal renin-angiotensin system (RAS) components and inflammatory mediators in diabetic rats. MAIN METHODS: Male Wistar rats (3 months old) were randomly assigned into four groups: non-trained (NT), trained (T), non-trained + diabetes (NTD) and trained +diabetes (TD). Diabetes was induced by streptozotocin (50 mg/kg, Sigma Chemical Co., St. Louis, MO, USA), before RT protocol. Trained rats performed RT protocol on a 110-cm ladder (8 ladder climbs, once/day, 5 days/week, 8 weeks), carrying a load corresponding to 50-80% of maximum carrying capacity. Blood glucose, albuminuria and urinary volume were measured. Renal levels of angiotensin peptides (angiotensin I, II and 1-7), inflammatory markers, and also the activities of angiotensin-converting enzyme (ACE) and ACE2 were determined. KEY FINDINGS: Blood glucose and urinary volume were elevated in diabetic animals, and RT decreased albuminuria, renal Ang I and Ang II levels in diabetic rats. RT shifted the balance of renal RAS toward ACE2/Ang 1-7 axis in TD group, and mitigated the high levels of interleukin (IL)-10, IL-1ß and cytokine-induced neutrophil chemoattractant 1 (CINC) in the context of diabetes. Strong positive correlations were found between albuminuria and Ang II, IL-10 and IL-1ß. On the other hand, intrarenal Ang 1-7 levels were negatively correlated with IL-10 and IL-1ß levels. SIGNIFICANCE: RT improved kidney function by modulating intrarenal RAS toward ACE2/Ang 1-7 axis and inflammatory cytokines. RT represents a reasonable strategy to improve the renal complications induced by diabetes, counteracting nephropathy-associated maladaptive responses.
Asunto(s)
Angiotensina I/metabolismo , Enzima Convertidora de Angiotensina 2/metabolismo , Diabetes Mellitus Experimental/metabolismo , Nefritis/metabolismo , Fragmentos de Péptidos/metabolismo , Sistema Renina-Angiotensina/fisiología , Entrenamiento de Fuerza/métodos , Animales , Diabetes Mellitus Experimental/terapia , Riñón/metabolismo , Masculino , Nefritis/terapia , Ratas , Ratas WistarRESUMEN
OBJECTIVES: Evaluation of the in-vivo anti-inflammatory activity of the methanolic extract obtained from the aerial parts of Mitracarpus frigidus (MFM) in the infection caused by two Salmonella strains and its chemical fingerprint by UFLC-quadrupole time of flight-MS. METHODS: The efficacy of MFM was investigated in a classical in-vivo Salmonella infection mouse model. A Salmonella reference strain (ATCC 13311) and a clinical isolate were used to infect mice and then MFM was orally administered during 14 days. At the end of the treatment with MFM, the infection and inflammatory levels were assayed. KEY FINDINGS: MFM treatment showed a significant reduction in mice mortality by Salmonella infection and, also, did not cause alterations in the liver function. Inhibitions of inflammatory and oxidative stress mediators [malondialdehyde (MDA), catalase, and metalloproteinase] were possibly involved in the observed effects. Chlorogenic acid, clarinoside, quercetin-pentosylhexoside, rutin, kaempferol-3O-rutinoside, kaempferol-rhamnosylhexoside and 2-azaanthraquinone were identified in MFM. CONCLUSIONS: MFM was effective in some inflammatory parameters, in the experimental conditions that were used in the study. The results presented in this study and the previous in-vitro anti-Salmonella activity reported by our research group reinforce the importance of MFM studies to considerer it as an alternative treatment for salmonellosis.
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Antiinflamatorios/uso terapéutico , Inflamación/prevención & control , Estrés Oxidativo/efectos de los fármacos , Fitoterapia , Extractos Vegetales/uso terapéutico , Rubiaceae/química , Infecciones por Salmonella , Animales , Antibacterianos/análisis , Antibacterianos/farmacología , Antibacterianos/uso terapéutico , Antiinflamatorios/análisis , Antiinflamatorios/farmacología , Antioxidantes/análisis , Antioxidantes/farmacología , Antioxidantes/uso terapéutico , Catalasa/metabolismo , Modelos Animales de Enfermedad , Inflamación/etiología , Inflamación/metabolismo , Hígado/efectos de los fármacos , Masculino , Malondialdehído/metabolismo , Metaloproteasas/metabolismo , Ratones , Fitoquímicos/análisis , Fitoquímicos/farmacología , Fitoquímicos/uso terapéutico , Componentes Aéreos de las Plantas , Extractos Vegetales/química , Extractos Vegetales/farmacología , Salmonella/efectos de los fármacos , Salmonella/crecimiento & desarrollo , Infecciones por Salmonella/complicaciones , Infecciones por Salmonella/tratamiento farmacológico , Infecciones por Salmonella/metabolismo , Infecciones por Salmonella/microbiología , Especificidad de la EspecieRESUMEN
Measurement of immunosuppressive drug concentrations cyclosporine A (CyA), tacrolimus (TAC), sirolimus (SIR), and everolimus (EVE) in blood is an important application of therapeutic drug monitoring. These immunosuppressive agents are used in combined regimens and nowadays the liquid chromatography and tandem mass spectrometry is the best option for simultaneous analysis of these drugs in one short run. We developed an liquid chromatography and tandem mass spectrometry methodology in-house to measure the combination of immunosuppressants in a single blood sample from transplant patients in Brazil. We analyzed 235 combinations of 4 immunosuppressive drugs in patient blood to validate this study. The measuring ranges were 9 to 1000 ng/mL for CyA and 2 to 50 ng/mL for TAC, SIR, and EVE. Accuracy of the method was between 83.87% and 126.6% (coefficient of determination [r2] > 0.995). Validation of variation was ≤15% for lower limit of quantification. In our analysis 20% of patients treated with EVE showed concentration range of 6 to 6.9 ng/mL, 28% of patients treated with SIR showed a concentration range of 4 to 4.9 ng/mL to TAC, 22% of patients showed concentration range of 5 to 5.9 ng/mL, and finally 50% of patients treated with CyA showed concentration range of 20 to 30 ng/mL. Our routine laboratory was able to implement this new methodology in-house to measure simultaneous CyA, TAC, SIR, and EVE in a single blood sample from transplant patients with a sensibility and rapid quantification analysis.
Asunto(s)
Cromatografía Liquida/métodos , Monitoreo de Drogas/métodos , Inmunosupresores/sangre , Espectrometría de Masas en Tándem/métodos , Brasil , Ciclosporina/sangre , Everolimus/sangre , Femenino , Rechazo de Injerto/prevención & control , Humanos , Trasplante de Órganos/efectos adversos , Sirolimus/sangre , Tacrolimus/sangreRESUMEN
The kidney is an important target of the renin-ANG-aldosterone system (RAAS). To date, several studies have demonstrated the existence of a local RAAS in various tissues, including the renal tissue. The mineralocorticoid aldosterone is known to play a critical role in the classical RAAS; however, its effect on mesangial cells (MCs) remains to be elucidated. Based on this, our aim was to investigate whether aldosterone stimulation can modulate the intracellular RAAS of immortalized human MCs by evaluating ANG-converting enzyme (ACE)/ANG II/ANG II receptor type 1 (AT1) and ANG-converting enzyme 2 (ACE2)/ANG (1-7)/MAS receptor axes. To realise this, protein expression, enzyme activity, and immunofluorescence were performed under aldosterone stimulation and in the presence of the mineralocorticoid receptor (MR) antagonist spironolactone (SPI). We observed that high doses of aldosterone increase ACE activity. The effect of aldosterone on the catalytic activity of ACE was completely abolished with the pretreatment of SPI suggesting that the aldosterone-induced cell injuries through ANG II release were attenuated. Aldosterone treatment also decreased the expression of MAS receptor, but did not alter the expression or the catalytic activity of ACE 2 and ANG (1-7) levels. Spironolactone modulated the localization of ANG II and AT1 receptor and decreased ANG (1-7) and MAS receptor levels. Our data suggest that both aldosterone and the MR receptor antagonist can modulate both of these axes and that spironolactone can protect MCs from the damage induced by aldosterone.
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Aldosterona/farmacología , Células Mesangiales/efectos de los fármacos , Células Mesangiales/metabolismo , Espironolactona/farmacología , Angiotensina I/genética , Angiotensina I/metabolismo , Enzima Convertidora de Angiotensina 2 , Células Cultivadas , Glicosilación/efectos de los fármacos , Humanos , Células Mesangiales/citología , Antagonistas de Receptores de Mineralocorticoides/farmacología , Fragmentos de Péptidos/genética , Fragmentos de Péptidos/metabolismo , Peptidil-Dipeptidasa A/genética , Peptidil-Dipeptidasa A/metabolismo , Proto-Oncogenes Mas , Proteínas Proto-Oncogénicas/genética , Proteínas Proto-Oncogénicas/metabolismo , Receptor de Angiotensina Tipo 1/genética , Receptor de Angiotensina Tipo 1/metabolismo , Receptores Acoplados a Proteínas G/genética , Receptores Acoplados a Proteínas G/metabolismo , Proteínas de Transporte Vesicular/genética , Proteínas de Transporte Vesicular/metabolismoRESUMEN
Diabetes mellitus and its complications have become a major health concern in Western countries. Increased activity of the intrarenal renin-angiotensin system (RAS) contributes to diabetic nephropathy (DN). We previously reported that in mesangial cells, the high glucose concentration (HG) leads to upregulation of angiotensin-converting enzyme (ACE) messenger RNA, suggesting that ACE was modulated by angiotensin II (Ang II) release. However, this relation in the collecting duct has not yet been studied. We, therefore, aimed to evaluate RAS modulation in inner medullary collecting duct cells (IMCD) exposed to HG. The IMCD were divided into normal glucose (5 mM D-glucose, NG), high glucose (30 mM, HG), and mannitol (30 mM, M) groups. The cells were cultured 48 hr in their respective media. The intracellular and extracellular ACE activity was measured using hippuryl-His-Leu as substrate via a fluorimetric assay and expression was analyzed using western blot analysis. ACE activity, intracellular (27%) and extracellular (22%), was significantly lower in the HG group than in NG and M. ACE2 activity and Ang 1-7 levels were higher in the intracellular compartment. Our data suggest that the HG cannot modify ACE synthesis in IMCD cells but can modulate its activity. The decrease in ACE activity may result in decreased levels of Ang II to protect the IMCD against proliferative and inflammatory deleterious effects of this peptide. Conversely, the increase of ACE2 generating high levels of Ang 1-7, a vasodilator peptide, suggesting that this peptide can induce glucose uptake and protect cells against oxidative stress, which can elicit insulin resistance.
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Glucosa/toxicidad , Túbulos Renales Colectores/efectos de los fármacos , Sistema Renina-Angiotensina/efectos de los fármacos , Angiotensina I/metabolismo , Angiotensina II/metabolismo , Enzima Convertidora de Angiotensina 2 , Animales , Línea Celular , Túbulos Renales Colectores/metabolismo , Túbulos Renales Colectores/patología , Ratones , Fragmentos de Péptidos/metabolismo , Peptidil-Dipeptidasa A/metabolismoRESUMEN
To test the effects of chronic-stress on the cardiovascular system, the model of chronic mild unpredictable stress (CMS) has been widely used. The CMS protocol consists of the random, intermittent, and unpredictable exposure of laboratory animals to a variety of stressors, during 3 consecutive weeks. In this study, we tested the hypothesis that exposure to the CMS protocol leads to left ventricle microcirculatory remodeling that can be attenuated by angiotensin II receptor blockade. Male Sprague-Dawley rats were randomly assigned into four groups: Control, Stress, Control + losartan, and Stress + losartan (N = 6, each group, losartan: 20 mg/kg/day). The rats were euthanized 15 days after CMS exposure, and blood samples and left ventricle were collected. Rats submitted to CMS presented increased glycemia, corticosterone, noradrenaline and adrenaline concentration, and losartan reduced the concentration of the circulating amines. Cardiac angiotensin II, measured by high-performance liquid chromatography (HPLC), was significantly increased in the CMS group, and losartan treatment reduced it, while angiotensin 1-7 was significantly higher in the CMS losartan-treated group as compared with CMS. Histological analysis, verified by transmission electron microscopy, showed that rats exposed to CMS presented increased perivascular collagen and losartan effectively prevented the development of this process. Hence, CMS induced a state of microvascular disease, with increased perivascular collagen deposition, that may be the trigger for further development of cardiovascular disease. In this case, CMS fibrosis is associated with increased production of catecholamines and with a disruption of renin-angiotensin system balance, which can be prevented by angiotensin II receptor blockade.
RESUMEN
ETHNOPHARMACOLOGICAL RELEVANCE: Vernonia condensata Baker (Asteraceae) is traditionally used in South American Countries as an anti-inflammatory, analgesic and hepatoprotective. AIM OF THE STUDY: This study aimed to investigate the in vivo hepatoprotective and antioxidant, and the in vitro anti-inflammatory activities of the ethyl acetate partition (EAP) from the ethanolic extract of this medicinal plant leaves. MATERIALS AND METHODS: For the in vivo hepatoprotective activity, rats were pretreated orally for seven days with vehicle, silymarin 100mg/kg or EAP 50, 100 and 200mg/kg. Then, acetaminophen 3g/kg was also orally administrated. Animals were euthanatized 24h after the damage inducement. The levels of the serum enzymes ALT, AST and ALP were determined, as well as the triglycerides, total cholesterol and fractions. The antioxidant activity was evaluated by TBARS assay and by the measurement of glutathione reductase, superoxide dismutase and catalase activities in the rats liver tissue. The in vitro anti-inflammatory assay using Raw 264.7 cell line induced by lipopolysaccharide was conducted to verify EAP ability to inhibit pro-inflammatory cytokines. RESULTS: EAP was able to inhibit all the acute biochemical alterations caused by acetaminophen overdose. EAP inhibited malondialdehyde formation, maintained the catalase and increased the glutathione reductase activities. Also, EAP decreased NO, IL-6 and TNF-α levels at concentrations from 10 to 20µg/mL. 1,5-dicaffeoylquinic acid was isolated and identified as the major compound in EAP. Apigenin, luteolin, chlorogenic acid were also identified. EAP anti-inflammatory action may be due to its antioxidant activity or its capacity to inhibit the pro-inflammatory cytokines. CONCLUSION: These results strongly suggested that V. condensata may be useful as a possible therapy against liver damage.
Asunto(s)
Antiinflamatorios/farmacología , Antioxidantes/farmacología , Enfermedad Hepática Inducida por Sustancias y Drogas/prevención & control , Extractos Vegetales/farmacología , Vernonia/química , Acetaminofén/administración & dosificación , Acetaminofén/toxicidad , Animales , Antiinflamatorios/administración & dosificación , Antiinflamatorios/aislamiento & purificación , Anticolesterolemiantes/administración & dosificación , Anticolesterolemiantes/aislamiento & purificación , Anticolesterolemiantes/farmacología , Antioxidantes/administración & dosificación , Antioxidantes/aislamiento & purificación , Línea Celular , Enfermedad Hepática Inducida por Sustancias y Drogas/etiología , Citocinas/metabolismo , Relación Dosis-Respuesta a Droga , Sobredosis de Droga , Macrófagos/efectos de los fármacos , Macrófagos/metabolismo , Masculino , Ratones , Extractos Vegetales/administración & dosificación , Hojas de la Planta , Ratas , Ratas Wistar , Silimarina/farmacologíaRESUMEN
Sepsis is an uncontrolled systemic inflammatory response against an infection and a major public health issue worldwide. This condition affects several organs, and, when caused by Gram-negative bacteria, kidneys are particularly damaged. Due to the importance of renin-angiotensin system (RAS) in regulating renal function, in the present study, we aimed to investigate the effects of endotoxemia over the renal RAS. Wistar rats were injected with Escherichia coli lipopolysaccharide (LPS) (4 mg/kg), mimicking the endotoxemia induced by Gram-negative bacteria. Three days after treatment, body mass, blood pressure, and plasma nitric oxide (NO) were reduced, indicating that endotoxemia triggered cardiovascular and metabolic consequences and that hypotension was maintained by NO-independent mechanisms. Regarding the effects in renal tissue, inducible NO synthase (iNOS) was diminished, but no changes in the renal level of NO were detected. RAS was also highly affected by endotoxemia, since renin, angiotensin-converting enzyme (ACE), and ACE2 activities were altered in renal tissue. Although these enzymes were modulated, only angiotensin (ANG) II was augmented in kidneys; ANG I and ANG 1-7 levels were not influenced by LPS. Cathepsin G and chymase activities were increased in the endotoxemia group, suggesting alternative pathways for ANG II formation. Taken together, our data suggest the activation of noncanonical pathways for ANG II production and the presence of renal vasoconstriction and tissue damage in our animal model. In summary, the systemic administration of LPS affects renal RAS, what may contribute for several deleterious effects of endotoxemia over kidneys.
Asunto(s)
Lesión Renal Aguda/metabolismo , Angiotensina II/metabolismo , Endotoxemia/metabolismo , Riñón/metabolismo , Lesión Renal Aguda/inducido químicamente , Lesión Renal Aguda/patología , Angiotensina I/metabolismo , Enzima Convertidora de Angiotensina 2 , Animales , Endotoxemia/inducido químicamente , Endotoxemia/patología , Riñón/patología , Lipopolisacáridos , Masculino , Óxido Nítrico/metabolismo , Óxido Nítrico Sintasa de Tipo II/metabolismo , Peptidil-Dipeptidasa A/metabolismo , Ratas , Ratas Wistar , Renina/metabolismo , Sistema Renina-Angiotensina/fisiologíaRESUMEN
OBJECTIVES: To evaluate the in-vivo wound healing and in-vitro antioxidant activity of gels containing the ethyl acetate extract of Cecropia pachystachya (ECP) 2% and 5%, and to perform the chemical fingerprint of ECP by HPLC-DAD. METHODS: The wound healing activity of the gels was evaluated for 21 days, using the excision model in rats followed by histopathological and histomorphometric analysis. The in-vitro antioxidant effect of ECP was investigated using 1-diphenyl-2-picrylhydrazyl, reducing power, ß-carotene bleaching and thiobarbituric acid reactive substances assays. Also, a HPLC analysis was performed to identify the chemical markers orientin, iso-orientin and chlorogenic acid. KEY FINDINGS: The group of animals treated with ECP 5% presented oedema and inflammatory infiltrate with less intensity than the other groups. Both ECP 2% and 5% gels showed less neovascularization and cellularity, and better tissue repair when compared to the control, which showed a younger and homogeneous tissue. CONCLUSIONS: This study had demonstrated that the ECP gels promoted the acceleration of the healing process when compared to the control group. Wound contraction, angiogenesis, epithelialization and the collagen deposition support further evaluation of C. pachystachya leaves in the topical treatment and management of skin wounds.
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Cecropia/química , Geles/química , Geles/farmacología , Extractos Vegetales/farmacología , Hojas de la Planta/química , Cicatrización de Heridas/efectos de los fármacos , Animales , Antioxidantes/química , Antioxidantes/farmacología , Compuestos de Bifenilo/química , Ácido Clorogénico/química , Flavonoides/química , Glucósidos/química , Masculino , Extractos Vegetales/química , Ratas , Ratas Wistar , Sustancias Reactivas al Ácido Tiobarbitúrico/químicaRESUMEN
Population studies have shown an association between diabetic nephropathy (DN) and insertion/deletion (I/D) polymorphism of the angiotensin-converting enzyme (ACE) gene (ACE in humans, Ace in mice). The aim was to evaluate the modulation of Ace copies number and diabetes mellitus (DM) on renal RAS and correlate it with indicators of kidney function. Increased number of copies of the Ace gene, associated with DM, induces renal dysfunction. The susceptibility to the development of DN in 3 copies of animals is associated with an imbalance in activity of RAS enzymes leading to increased synthesis of Ang II and Ang-(1-7). Increased concentration of renal Ang-(1-7) appears to potentiate the deleterious effects triggered by Ang II on kidney structure and function. Results also show increased bradykinin concentration in 3 copies diabetic group. Taken together, results indicate that the deleterious effects described in 3 copies diabetic group are, at least in part, due to a combination of factors not usually described in the literature. Thus, the data presented here show up innovative and contribute to understanding the complex mechanisms involved in the development of DN, in order to optimize the treatment of patients with this complication.
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Angiotensina I/metabolismo , Bradiquinina/metabolismo , Nefropatías Diabéticas/genética , Regulación de la Expresión Génica , Riñón/metabolismo , Fragmentos de Péptidos/metabolismo , Animales , Glucemia/análisis , Peso Corporal , Diabetes Mellitus Experimental/metabolismo , Diabetes Mellitus Tipo 1/complicaciones , Nefropatías Diabéticas/metabolismo , Dosificación de Gen , Predisposición Genética a la Enfermedad , Genotipo , Masculino , Ratones , Ratones Endogámicos C57BL , Peptidil-Dipeptidasa A/genética , Sistema Renina-Angiotensina/efectos de los fármacos , Espectrometría de FluorescenciaRESUMEN
This study aimed to analyzing the effect of chronic sodium overload upon carotid and femoral injury, and its relation to vascular angiotensin modulation. Male C57Bl6 mice were divided in: control (cont), receiving 1% NaCl solution for 2 weeks (salt-2) or 12 weeks (salt-12). Two-weeks before the end of the study, a 2mm catheter was implanted around the left femoral and carotid arteries to induce injury. Blood pressure (BP) and heart rate (HR) were measured at the end of the study by tail plethysmography. Arteries were collected and prepared for histological analysis to determine arterial thickening and perivascular collagen deposition. Angiotensin II and Ang(1-7) were quantified in fresh arteries using the HPLC method. There were no differences in body weight, BP and HR. Intima/media ratio had a similar increase in both injured arteries of cont and salt-2 mice, but a more pronounced increase was observed in salt-12 mice (31.1±6%). On the other hand, sodium overload modified perivascular collagen deposition, increasing thick fibers (cont: 0.5%; salt-2: 3.4%; salt-12: 0.6%) and decreasing thin fibers (cont: 7.4%; salt-2: 0.5%; salt-12: 6.8%) in non-injured arteries. Injured arteries presented similar collagen fiber distribution. Angiotensin quantification showed increased Ang(1-7) in salt treated mice (salt-2: +72%; salt-12: +45%) with a concomitant decrease in Ang II (salt-2: -54%; salt-12: -60%). Vascular injury increased significantly Ang(1-7) in salt-12 mice (+80%), maintaining Ang II reduction similar to that of a non-injured artery. The lack of changes in BP and HR suggests that the structural changes observed may be due to non-hemodynamic mechanisms such as local renin-angiotensin system. Collagen evaluation suggests that sodium overload induces time-related changes in vascular remodeling. The increase of artery injury with concomitant increase in Ang(1-7) in 12-week treated mice shows a direct association between the duration of salt treatment and the magnitude of vascular injury.
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Angiotensinas/metabolismo , Arterias Carótidas/efectos de los fármacos , Traumatismos de las Arterias Carótidas/inducido químicamente , Arteria Femoral/efectos de los fármacos , Arteria Femoral/lesiones , Cloruro de Sodio/efectos adversos , Animales , Presión Sanguínea/efectos de los fármacos , Peso Corporal/efectos de los fármacos , Arterias Carótidas/metabolismo , Arterias Carótidas/patología , Arterias Carótidas/fisiología , Colágeno/metabolismo , Relación Dosis-Respuesta a Droga , Elasticidad , Arteria Femoral/patología , Arteria Femoral/fisiología , Frecuencia Cardíaca/efectos de los fármacos , Hemodinámica/efectos de los fármacos , Masculino , Ratones , Ratones Endogámicos C57BL , Neointima/inducido químicamente , Peptidil-Dipeptidasa A/metabolismo , Remodelación Vascular/efectos de los fármacosRESUMEN
BACKGROUND: Hyperactivity of the renin-angiotensin system (RAS) and functional deficits in hypertension are reduced after exercise training. We evaluate in arteries, kidney and plasma of hypertensive rats the sequential effects of training on vascular angiotensinogen, Ang II and Ang (1-7) content. METHODS AND RESULTS: Spontaneously hypertensive rats (SHR) and Wistar-Kyoto rats (WKY) were trained or kept sedentary (S) for 3 months. After hemodynamic measurements (weeks 0, 1, 2, 4, 8 and 12), blood, arteries and kidneys were obtained to quantify the angiotensin content (HPLC) and angiotensinogen expression (Western Blotting). SHR-S vs. WKY-S exhibited elevated pressure, increased angiotensinogen and angiotensins' content in the renal artery with a high Ang II/Ang (1-7) ratio (~5-fold higher than in the femoral artery, kidney and plasma, and 14-fold higher than in the aorta). Training promptly reduced angiotensinogen expression and downregulated the RAS in the renal SHR artery (1st-12th week), with a specific reduction of the vasoconstrictor axis; significant reduction of the AngII/Ang (1-7) ratio (36%, T4-T8) occurred simultaneously with significant pressure fall (5%). In other SHR arteries, plasma and kidneys and in all WKY tissues, T-induced AngII and Ang (1-7) reductions were proportional, maintaining the AngII/Ang (1-7) ratio. CONCLUSIONS: Vascular RAS is not equally expressed in vessels, having crucial importance in the renal artery. In the renal SHR artery, training downregulates the vasoconstrictor and preserves the vasodilator axis while in other tissues and plasma training reduces both RAS axes, thus maintaining the vasoconstriction/vasodilatation balance in a lower level.
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Angiotensina II/biosíntesis , Angiotensina I/biosíntesis , Angiotensinógeno/biosíntesis , Riñón/metabolismo , Fragmentos de Péptidos/biosíntesis , Condicionamiento Físico Animal/fisiología , Arteria Renal/metabolismo , Sistema Renina-Angiotensina/fisiología , Aerobiosis/fisiología , Angiotensina I/sangre , Angiotensina II/sangre , Enzima Convertidora de Angiotensina 2 , Angiotensinógeno/sangre , Angiotensinógeno/genética , Animales , Presión Sanguínea , Arteria Femoral , Masculino , Especificidad de Órganos , Fragmentos de Péptidos/sangre , Peptidil-Dipeptidasa A/sangre , Ratas , Ratas Endogámicas SHR , Ratas Endogámicas WKY , Carrera , Vasoconstricción/fisiología , Vasodilatación/fisiologíaRESUMEN
Somatic angiotensin-I converting enzyme (sACE) is a broadly distributed peptidase which plays a role in blood pressure and electrolyte homeostasis by the conversion of angiotensin I into angiotensin II. N-domain isoforms (nACE) with 65 and 90 kDa have been described in body fluids, tissues and mesangial cells (MC), and a 90 kDa nACE has been described only in spontaneously hypertensive rats. The aim of this study was to investigate the existence of proteolytic enzymes that may act in the hydrolysis of sACE generating nACEs in MC. After the confirmation of the presence of ACE sheddases in Immortalized MC (IMC), we purified and characterized these enzymes using fluorogenic substrates specifically designed for ACE sheddases. Purified enzyme identified as a serine protease by N-terminal sequence was able to generate nACE. In the present study, we described for the first time the presence of ACE sheddases in IMC, identified as serine proteases able to hydrolyze sACE in vitro. Further investigations are necessary to elucidate the mechanisms responsible for the expression and regulation of ACE sheddases in MC and their roles in the generation of nACEs, especially the 90 kDa form possibly related to hypertension.
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Hipertensión/enzimología , Peptidil-Dipeptidasa A/metabolismo , Proteolisis , Serina Proteasas/metabolismo , Angiotensina I/metabolismo , Angiotensina II/metabolismo , Animales , Presión Sanguínea , Humanos , Hipertensión/metabolismo , Células Mesangiales/enzimología , Células Mesangiales/metabolismo , Peptidil-Dipeptidasa A/biosíntesis , Isoformas de Proteínas/biosíntesis , Isoformas de Proteínas/metabolismo , Ratas , Serina Proteasas/biosíntesisRESUMEN
It has been clearly established that mitogen-activated protein kinases (MAPKS) are important mediators of angiotensin II (Ang II) signaling via AT1 receptors in the vasculature. However, evidence for a role of these kinases in changes of Ang II-induced vasoconstriction in obesity is still lacking. Here we sought to determine whether vascular MAPKs are differentially activated by Ang II in obese animals. The role of AT2 receptors was also evaluated. Male monosodium glutamate-induced obese (obese) and non-obese Wistar rats (control) were used. The circulating concentrations of Ang I and Ang II, determined by HPLC, were increased in obese rats. Ang II-induced isometric contraction was decreased in endothelium-intact resistance mesenteric arteries from obese compared with control rats and exhibited a retarded AT1 receptor antagonist response. Blocking of AT2 receptors and inhibition of either endothelial nitric oxide synthase (eNOS) or extracellular signal-regulated protein kinases 1 and 2 (ERK1/2) restored Ang II-induced contraction in obese rats. Western blot analysis revealed increased protein expression of AT2 receptors in arteries from obese rats. Basal and Ang II-induced ERK1/2 phosphorylation was also increased in obese rats. Blockade of either AT1 or AT2 receptors corrected the increased ERK1/2 phosphorylation in arteries from obese rats to levels observed in control preparations. Phosphorylation of eNOS was increased in obese rats. Incubation with the ERK1/2 inhibitor before Ang II stimulation did not affect eNOS phosphorylation in control rats; however, it corrected the increased phosphorylation of eNOS in obese rats. These results clearly demonstrate that enhanced AT2 receptor and ERK1/2-induced, NO-mediated vasodilation reduces Ang II-induced contraction in an endothelium-dependent manner in obese rats.
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Arterias Mesentéricas/fisiología , Proteína Quinasa 1 Activada por Mitógenos/metabolismo , Proteína Quinasa 3 Activada por Mitógenos/metabolismo , Óxido Nítrico Sintasa de Tipo III/metabolismo , Obesidad/fisiopatología , Receptor de Angiotensina Tipo 2/metabolismo , Angiotensina II/farmacología , Bloqueadores del Receptor Tipo 2 de Angiotensina II/farmacología , Animales , Western Blotting , Endotelio Vascular/fisiología , Inhibidores Enzimáticos/farmacología , Imidazoles/farmacología , Técnicas In Vitro , Masculino , Arterias Mesentéricas/metabolismo , Proteína Quinasa 1 Activada por Mitógenos/antagonistas & inhibidores , Proteína Quinasa 3 Activada por Mitógenos/antagonistas & inhibidores , NG-Nitroarginina Metil Éster/farmacología , Óxido Nítrico/metabolismo , Óxido Nítrico Sintasa de Tipo III/antagonistas & inhibidores , Obesidad/metabolismo , Fosforilación/efectos de los fármacos , Piridinas/farmacología , Ratas Wistar , Regulación hacia Arriba , Vasoconstricción/efectos de los fármacos , Vasoconstrictores/farmacologíaRESUMEN
The aims of this work were to evaluate the in vitro and in vivo schistosomicidal properties of the methanolic extract of the aerial parts of Mitracarpus frigidus (MFM) and to determine its HPLC profile. For the in vitro experiment, four pairs of adult worms, obtained from infected mice, were exposed to different concentrations of MFM (100 to 400 µg/mL) for 24 and 48 h and analyzed under an inverted microscope. For the in vivo experiment, mice were inoculated with cercariae and, 20 days after infection, MFM (100 and 300 mg/kg) was administered orally for the following 25 days. Mice were euthanized after 60 days. MFM showed in vitro schistosomicidal activity, exhibiting the opening of the gynaecophoral canal of some male schistosomes, the presence of contorted muscles, vesicles, and the darkening of the paired worms skin. In vivo experiments showed that MFM treatments significantly reduced total worm count, as praziquantel, showing a decrease in liver and spleen weight. Also, a significant reduction in granuloma density was observed. MFM treatment did not cause alterations in the liver function of either infected or noninfected mice. The HPLC chromatogram profile showed the presence of kaempferol-O-rutinoside, rutin, kaempferol, psychorubrin, and ursolic acid.
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Extractos Vegetales/farmacología , Plantas Medicinales/química , Rubiaceae/química , Schistosoma mansoni/efectos de los fármacos , Animales , Cromatografía Líquida de Alta Presión/métodos , Femenino , Granuloma/tratamiento farmacológico , Quempferoles/química , Hígado/efectos de los fármacos , Masculino , Ratones , Naftoquinonas/química , Extractos Vegetales/química , Rutina/química , Esquistosomicidas/farmacología , Bazo/efectos de los fármacos , Triterpenos/química , Ácido UrsólicoRESUMEN
Cecropia pachystachya is a species traditionally used in Brazil to treat inflammation. This work aims to evaluate the topical anti-inflammatory and antioxidant activities of the methanolic extract of C. pachystachya (CPM) and to perform its chemical fingerprint by HPLC-DAD. The topical anti-inflammatory activity was evaluated using the mouse models of acute ear inflammation induced by croton oil, arachidonic acid, capsaicin, EPP, phenol, and chronic inflammation induced by multiple application of croton oil. The in vitro antioxidant effect of CPM was investigated using DPPH, reducing power, ß -carotene bleaching, and TBARS assays. HPLC analysis was performed to quantify the antioxidant phenolics orientin, isoorientin, and chlorogenic acid previously identified in CPM. CPM exhibited significant anti-inflammatory effect in the acute models, in some cases comparable to the reference drugs. Histopathological analysis showed a moderate chronic skin anti-inflammatory effect with decrease in vasodilation, edema, cell infiltration, and epidermal hyperproliferation. It also showed strong in vitro antioxidant activity. The contents of orientin, isoorientin, and chlorogenic acid were 66.5 ± 1.8, 118.8 ± 0.7, and 5.4 ± 0.2 µg/mg extract, respectively. The topical anti-inflammatory activity of CPM could be based on its antioxidant properties, although other effects are probably involved, including COX inhibition and other mechanisms.
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Antiinflamatorios/farmacología , Antioxidantes/farmacología , Cecropia/química , Dermatitis/tratamiento farmacológico , Enfermedades del Oído/tratamiento farmacológico , Extractos Vegetales/farmacología , Administración Tópica , Animales , Antiinflamatorios/química , Antioxidantes/química , Dermatitis/metabolismo , Dermatitis/patología , Modelos Animales de Enfermedad , Enfermedades del Oído/inducido químicamente , Enfermedades del Oído/metabolismo , Enfermedades del Oído/patología , Epidermis/metabolismo , Epidermis/patología , Masculino , Ratones , Extractos Vegetales/químicaRESUMEN
The present study evaluated the antioxidant potential of Vernonia condensata Baker (Asteraceae). Dried and powdered leaves were exhaustively extracted with ethanol by static maceration followed by partition to obtain the hexane, dichloromethane, ethyl acetate, and butanol fractions. Total phenols and flavonoids contents were determined through spectrophotometry and flavonoids were identified by HPLC-DAD system. The antioxidant activity was assessed by DPPH radical scavenging activity, TLC-bioautography, reducing power of Fe(+3), phosphomolybdenum, and TBA assays. The total phenolic content and total flavonoids ranged from 0.19 to 23.11 g/100 g and from 0.13 to 4.10 g/100 g, respectively. The flavonoids apigenin and luteolin were identified in the ethyl acetate fraction. The IC50 of DPPH assay varied from 4.28 to 75.10 µg/mL and TLC-bioautography detected the antioxidant compounds. The reducing power of Fe(+3) was 19.98 to 336.48 µg/mL, while the reaction with phosphomolybdenum ranged from 13.54% to 32.63% and 56.02% to 135.00% considering ascorbic acid and rutin as reference, respectively. At 30 mg/mL, the ethanolic extract and fractions revealed significant effect against lipid peroxidation. All these data sustain that V. condensata is an important and promising source of bioactive substances with antioxidant activity.