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- ItemEfeitos da sobrecarga crônica de ferro sobre a estrutura e função de artéria coronária de ratos(Universidade Federal do Espírito Santo, 2023-09-29) Rodrigues, Sabrina Bertoli; Santos, Leonardo dos; https://orcid.org/0000-0002-4340-6364; http://lattes.cnpq.br/4132087001362623; https://orcid.org/0000-0003-1134-5349; http://lattes.cnpq.br/8927452634782526; Bissoli, Nazaré Souza; https://orcid.org/0000-0002-3456-2437; http://lattes.cnpq.br/8865368585732583; Santos, Paulo Caleb Júnior de Lima; http://lattes.cnpq.br/7270343730265469; Furieri, Lorena Barros; https://orcid.org/0000000338592227; http://lattes.cnpq.br/0526203649991750; Rossoni, Luciana VenturiniIron is an essential mineral for several cellular processes, mainly due toitsabilitytoparticipate in reactions in which it receives or donates electrons. For thesamereason, when free and in excess, it is a great precursor in the generationof reactiveoxygen species, damaging tissues, organs and systems. Several studiesdemonstrate that iron overload is capable of causing cardiovascular damage, andaprobable relationship between the level of body iron and coronary artery diseasehaseven been suggested. In this study, we aimed to test the hypothesis that chronicironoverload, per se, causes damage to the coronary vasculature, associatedwiththegenerated oxidative stress and endothelial dysfunction. Serum, tissues andcoronaryvessels of male Wistar rats from the Ct and Fe groups were analyzed, inwhichsalinesolution (NaCl 0.9%) or iron-dextran (200 mg/kg/day) were administeredintraperitoneally, respectively, 5 times a week for 28 days. After euthanasia, bloodand organs were collected to assess serum and tissue iron, and vasoreactivity,fluorescence for reactive oxygen species and nitric oxide, histolomorphometrywereperformed in isolated coronary arteries. Finally, the modified Langedorff techniquetoassess the coronary bed was performed in a new set of experimental rats. Asexpected, iron overload increased serum and tissue iron levels, as well asreducedthe weight gain of animals in the Fe group compared to the Ct group. Furthermore,there was an increase in reactivity and a reduction in the vasodilator responseintheisolated rings of coronary arteries in the Fe group, associated with an increaseinthegeneration of superoxide anion, probably mediated by the AT1 receptor. Ironoverloadalso significantly reduced the bioavailability of nitric oxide in the coronary arteries. Inaddition to functional alterations, remodeling of arteries in the irongroupwasobserved, with collagen deposits and the presence of perivascular macrophages;together with this, alteration of the vascular endothelium with cell detachment wasevidenced, which suggests a denudation of this layer. In the analysis of theisolatedcoronary vascular bed, an increase in coronary perfusion pressure was observedinthis same Fe group, probably as a consequence of the increaseinvascularresistance and vasculopathy from iron overload. Our results demonstratethat chroniciron overload induces coronary endothelial dysfunction, probably duetooxidativestress and the imbalance between relaxing and contractile factors synthesizedbythedamaged endothelium.
- ItemO antioxidante mitocondrial, mitoquinona, previne a disfunção cardíaca 7 dias após infarto do miocárdio em rato(Universidade Federal do Espírito Santo, 2023-08-08) Ximenes, Carolina Falcão; Stefanon, Ivanita; https://orcid.org/0000-0003-2638-5183; http://lattes.cnpq.br/8456612999765726; https://orcid.org/0000-0001-6619-2285; http://lattes.cnpq.br/3613329548109549; Bissoli, Nazaré Souza; https://orcid.org/0000-0002-3456-2437; http://lattes.cnpq.br/8865368585732583; Baldo, Thaís de Oliveira Faria; https://orcid.org/0000-0001-8391-6177; http://lattes.cnpq.br/2809630268648026; Fonseca, Silvia Carolina Guatimosim; https://orcid.org/0000-0001-8386-3722; http://lattes.cnpq.br/7958786029463633Myocardial Infarction (AMI) is considered the main cause of heart failure (HF). It is believed that oxidative stress (ROS) plays a crucial role in the myocardium adjacent to the infarcted area and in the progression of left ventricular remodeling. At 7 days post-AMI, the decrease in myocardial contractility is associated with changes in the pathway of calcium (Ca2+) and ROS. However, the impact of mitochondrial ROS as a source of controlled dysfunction during the early phase of AMI will remains unknown. We hypothesize that MitoQ mitochondrial antioxidant treatment for 7 days after AMI will improve contractile function dependent on the reduction in mitochondrial ROS production in the acute phase of AIM. Therefore, our objective was to analyze the effect of treatment, for 7 days, with the specific mitochondrial antioxidant, MitoQ, on contractile dysfunction in the acute phase after AMI in rats. Wistar rats aged 12 weeks were divided into Sham, Infarto, Sham MitoQ and Infarto MitoQ (CEUA 16/2021). At the end of treatment with MitoQ for 7 days in drinking water (100 µM), analyzes of myocardial contractility “in vivo” were performed in hemodynamic parameters and “in vitro” measured in isolated papillary left ventricle (LV) with muscle length in which the active voltage is maximum (Lmáx); in presence of different extracellular concentrations of MitoQ; Ca2+ and isoproterenol. Cardiomyocytes isolated from the LV were used to measure morphological and temporal parameters of contractile function and transient Ca2+. Superoxide anion (O2 •- ) production was quantified using Dihydroethidium (DHE) and mitochondrial O2 •- using MitoSox Red. Statistical analysis used ANOVA two-way and post-hoc de Tukey’s and test t student, for p<0,05. Treatment with MitoQ did not change the area of infarction, However, it prevented the decrease in body weight gain and prevented the hemodynamic changes observed in the infarction group in the following parameters: systolic blood pressure (SBP) Sham: 113 ± 3; Infarto: 93 ± 4; Sham MitoQ: 108 ± 4; Infarto MitoQ: 108 ± 3*, mmHg *p<0.05); diastolic blood pressure (DBP) (Sham: 84 ± 3; Infarto: 68 ± 3; Sham MitoQ: 81 ± 3; Infarto MitoQ: 108 ± 3*, mmHg*p<0.05); left ventricular systole pressure (LVSP) (Sham: 116 ± 4.5; Infarto: 79 ± 2.7; Sham MitoQ: 115.8 ± 6; Infarto MitoQ: 100 ± 4.4*, mmHg *p<0.05); left ventricular end-diastolic pressure (LVDP) (Sham: 5 ± 0.4; Infarto: 10 ± 1; Sham MitoQ: 3 ± 1; Infarto MitoQ: 5 ± 0.5*, mmHg *p<0.05); first derivate of maximum pressure (dP/dtmáx) (Sham: 3993 ± 199; Infarto: 2480 ± 58; Sham MitoQ: 3326 ± 265; Infarto MitoQ: 2738 ± 103*, mmHg/s *p<0.05) and first derivate of minimum (dP/dtmin) (Sham: -3273 ± 227; Infarto: -1486 ± 41; Sham MitoQ: -2703 ± 88; Infarto MitoQ: -2083 ± 88*, mmHg/s *p<0,05). Treatment with MitoQ (100 uM) for 7 days was able to prevent the reduction of the isometric force of contraction of the animals in the infarction group (Sham: 0.56 ± 0.06; Infarto: 0.29 ± 0.05; Sham MitoQ: 0.47 ± 0.07; Infarto MitoQ: 0.73 ± 0.08* g/mg, *p<0.05), the reduction of the maximum positive derivative of force (+dF/dtmáx) (Sham: 25 ± 2.18; Infarto: 14.43 ± 2.20; Sham MitoQ: 16.88 ± 2.16; Infarto MitoQ: 24.17 ± 1.75 g/g/s, *p<0.05) and reduced contractility to extracellular Ca2+ influx (1,25 mM – Sham: 444 ± 49.86; Infarto: 229.83 ± 68.28; Sham MitoQ: 357.38 ± 29.97; Infarto MitoQ: 640.17 ± 60.9* g/g, *p<0.05). Morphometric analyses observed that treatment with MitoQ prevented the increase in cell area (Sham: 3846 ± 105; Infarto: 4309 ± 107; Sham MitoQ: 4107 ± 134; Infarto MitoQ: 3782 ± 116* µm2 , *p<0.05) and increased cell length (Sham: 141 ± 2.8; Infarto: 153 ± 1.95; Sham MitoQ: 144 ± 2.25; Infarto MitoQ: 137 ± 2.25* µm, *p<0,05) of cardiomyocytes in the initial stage of AMI. The cardiomyocytes contractility was increased in the Infarto group, which were prevented by MitoQ treatment such as cardiomyocyte shortening (Sham: 610 ± 26; Infarto: 815 ± 30; Sham MitoQ: 653,5 ± 25; Infarto MitoQ: 553,6 ± 30* µm2 , *p<0.05). MitoQ treatment prevented the increase in [Ca2+]i transient amplitude (Sham: 2.58 ± 0.04; Infarto: 3.15 ± 0.07; Sham MitoQ: 2.65 ± 0.08; Infarto MitoQ: 2.30 ± 0.05* F/F0, *p<0.05). The in-situ production of O2 •- , demonstrated by fluorescence intensity, was higher in the Infarto group, but treatment with MitoQ for 7 days prevented this increase in ROS. Mitochondrial O2 •- formation in isolated cardiomyocytes was greater in the Infarto group compared to the Sham group (p<0.01), and MitoQ treatment restored redox homeostasis. Our results demonstrated that MitoQ treatment prevented contractile dysfunction, confirming the involvement of mitochondrial ROS participation in the development of HF after AMI. In this way, find the most effective and safe way to modulate mitochondrial function and dynamics in HF after AMI in its initial phase represents a potential therapeutic target and an important step for the future of research in the treatment of cardiovascular diseases.
- ItemUso da espectroscopia FT-IR ATR e métodos de quimiometria em biofluidos para diagnósticos clínicos: efeito da diluição da amostra no diagnóstico da doença de Fabry(Universidade Federal do Espírito Santo, 2023-05-30) Leal, Leonardo Barbosa; Barauna, Valerio Garrone; https://orcid.org/0000000328320922; http://lattes.cnpq.br/1583882564447423; https://orcid.org/0000-0003-0968-4202; http://lattes.cnpq.br/1630453093149668; Graeff, Carlos Frederico de Oliveira; https://orcid.org/0000-0003-0162-8273; http://lattes.cnpq.br/5268607684223281; Mill, Jose Geraldo; https://orcid.org/000000020987368X; http://lattes.cnpq.br/2497419234600362; Teixeira, Carlos Graeff; https://orcid.org/0000000327250061; http://lattes.cnpq.br/0464152494769261; Valderrama, Patricia; Lima, Kassio Michell Gomes deFourier Transform Infrared Spectroscopy (ATR-FTIR) emerges as a powerful tool to aid the area of clinical medicine. The FTIR technique provides accurate, real-time results for clinical/diagnostic screening tests. This study aimed to study the effect of plasma sample dilution on Fabry disease classification. Fabry disease (FD), a rare X cromossome disorder with alpha-galactosidase A (α-GAL-A) deficiency. The equipment used was the Agilent Cary 630 spectrometer, equipped with a diamond crystal, Total Attenuated Reflection (ATR) system. The range between 400 to 4000 cm-1 was analyzed with a resolution of 4cm-1 with 32 raster and background scans. Analyzes of volume, drying time and dilution were carried out. Once the best dilution point was located, plasma samples from patients with SCD were analyzed in their total and diluted form. Average spectra in triplicates were obtained and analyzed in separate regions, FG fingerprint (800-1800cm-1 ) and high HW wavenumber (2800-4000cm-1 ) and pre-processed by line vector normalization. Unsupervised analyzes were performed, such as principal component analysis (PCA) and Unsupervised Random Forest (URF) and classification analysis such as GA-LDA along with Monte Carlo Method with 500 models, Support Vector Machine (SVM), K-near Neighbor (KNN) and PLS-DA (Partial Least Squres – Discriminated Analysis) were used to obtain the results. The results suggested that the ideal drying time for plasma analysis is from 18 minutes and a volume of 10µl. For dilution, the best result was 25%. Both models built with diluted and undiluted samples were able to identify patients with SCD with specificity and sensitivity above 80%. Models built with undiluted samples were able to generate the best predictive model for PD. However, it is still necessary to investigate the effect of dilution in the making of predictive models created from other biological samples, such as, for example, urine.
- ItemEfeitos da testosterona sobre a reatividade vascular de ratos hipertensos(Universidade Federal do Espírito Santo, 2023-02-15) Gonçalves, Leticia Tinoco; Santos, Roger Lyrio dos; https://orcid.org/0000-0003-3399-3523; http://lattes.cnpq.br/1122196233280741; https://orcid.org/0000-0003-2202-8541; http://lattes.cnpq.br/0836713525136907; Ceravolo, Graziela Scalianti; https://orcid.org/0000-0001-7558-6294; http://lattes.cnpq.br/0517142252318597; Padilha, Alessandra Simao; https://orcid.org/0000-0002-9585-1347; http://lattes.cnpq.br/7658998034219799; Bonaventura, Daniella; https://orcid.org/0000-0001-5253-4918; http://lattes.cnpq.br/9290907947235226; Santos, Leonardo dos; http://lattes.cnpq.br/4132087001362623The abusive use of testosterone is considered a global public health problem, with several side effects. In the cardiovascular system, its effects are still controversial, ranging from protective to deleterious actions. Because testosterone is a hormone that can be converted to 17 β-estradiol and dihydrotestosterone (DHT), part of the effects found with supraphysiological doses can be attributed to its metabolites. Therefore, we investigated the hypothesis that a supraphysiological dose of testosterone impairs the endothelium-dependent vasodilation of mesenteric resistance arteries, as well as its repercussions on oxidative stress (OS) and blood pressure (BP). We also evaluated the participation of 17 β-estradiol and DHT in the responses found. We used Spontaneously Hypertensive Rats (SHR), aged 8 to 10 weeks, divided into 5 groups: intact (SHAM), orchiectomized (ORX), intact testosterone-treated (TTO; 3 mg/Kg/day/s.c.), intact treated with testosterone and anastrozole [aromatase enzyme inhibitor (TTO+ANA; 0.1 mg/Kg/day)] and intact treated with testosterone and finasteride [5α-reductase enzyme inhibitor (TTO+FIN; 5 mg/Kg/day)] for 4 weeks. BP was assessed by tail cuff plethysmography. We performed concentration-response curves to acetylcholine (ACh, 0.1 nM - 10 µM) in mesenteric arteries using a wire myograph, in the absence and presence of pharmacological inhibitors. Also testosterone, 17 β-estradiol and dihydrotestosterone concentrations were evaluated. Vascular detection of superoxide anion (O2 •- ) and endothelium ultrastructure were analyzed by DHE and scanning electron microscopy (SEM), respectively. Data were expressed as mean ± standard error of the mean, and analyzed by Student's t-test or one-way or two-way ANOVA, followed by Tukey's post hoc test (p < 0.05). Orchiectomy reduced levels of testosterone, 17 β-estradiol and dihydrotestosterone, impaired ACh vasodilation, increased OS, altered endothelial morphology without altering BP. Testosterone treatment did not impair ACh vasodilation compared to the SHAM group, however it altered the endothelial pathways of relaxation, with lesser participation of NO and greater participation of prostanoids, possibly derived from COX-1. In addition, in the TTO group, the participation of EDH was greater compared to SHAM, indicating that EETs, H2O2 and K+ channels contributed to this vasodilator response. In the TTO+ANA group, the reduction in 17 β-estradiol levels did not impair ACh vasodilation, however, it decreased the participation of nitric oxide, prostanoids and increased EDH, and increased O2•- levels with alteration of endothelial morphology. TTO+FIN showed impairment in the vasodilator response to ACh, with an increased participation of NO and a lower participation of prostanoids. Regarding EDH, with a decrease in dihydrotestosterone, there was no increase in the participation of EETs, H2O2 and K+ channels compared to TTO. DHT seems to contribute to the decrease of NO and estrogen seems to stimulate the action of the NO pathway and prostanoids. In estrogen reduction, testosterone maintains endothelial vasodilation by greater stimulation of EDH, with more action of EETs, H2O2 and K+ channels, with greater formation of O2 •- . These results may contribute to the elucidation of the modulating role of testosterone on endothelial function, even in treatment with a supraphysiological dose, in addition to showing the importance of the presence of estrogen for the cardiovascular system in situations of endothelial dysfunction.
- ItemPotencial toxicológico do cádmio em dose semelhante à de exposição ocupacional na gênese de complicações no trato reprodutivo de ratas wistar(Universidade Federal do Espírito Santo, 2023-02-16) Costa, Charles Santos da; Graceli, Jones Bernardes; https://orcid.org/0000000193494369; http://lattes.cnpq.br/3803670746263603; https://orcid.org/0000-0003-3616-8211; http://lattes.cnpq.br/8647659013623563; Alves, Leandro Miranda; https://orcid.org/0000-0001-7832-8717; http://lattes.cnpq.br/9883547704448526; Padilha, Alessandra Simão; https://orcid.org/0000-0002-9585-1347; http://lattes.cnpq.br/7658998034219799; Santos, Roger Lyrio dos; https://orcid.org/0000-0003-4316-7196; http://lattes.cnpq.br/1122196233280741; Lima, Leandro Ceotto FreitasCadmium (Cd) is a toxic heavy metal known to be a major endocrine disruptor. However, few studies have explored the effects of Cd exposure on developing polycystic ovary syndrome (PCOS) features and premature ovarian failure (POF). In this study, we evaluated whether subacute exposure to Cd at a dose similar to that of occupational exposure results in damage to the hypothalamic-pituitarygonadal (HPG) axis and abnormalities related to PCOS and FOP. Wistar rats were exposed to CdCl₂ (100 ppm in water for 30 days), we evaluated the levels of Cd in the blood, HPG axis and uterus. In addition, we evaluated metabolic factors, HPG axis function, reproductive tract morphophysiology, inflammation, oxidative stress and fibrosis. Exposure to Cd raised its levels in serum, HPG axis and uterus. The Cd rats presented metabolic damage, adiposity reduction, dyslipidemia and insulin resistance (IR). Cd also caused the HPG axis to malfunction. Irregularities in the estrous cycle, abnormal hypothalamic mRNA expression with overexpression of Kisspeptin 1 (Kiss1), androgen receptor (AR) and rapamycin mechanistic target protein (mTOR) and lower expression of Kiss1 (Kiss1R) and leptin receptors (LepR) were observed and tumor necrosis factor alpha (TNF-a), high levels of luteinizing hormone (LH), low levels of anti-Müllerian hormone (AMH) and abnormal ovarian follicular development, reduced ovarian reserve and follicle number antral were observed, suggesting ovarian depletion. Furthermore, exposure to Cd decreases the granulosa layer thickness and the corpora lutea counting and increases the cystic and atretic follicles. Cd exposure increased inflammation, oxidative stress, and remodeling in the reproductive tract. Finally, positive correlations were observed between serum and reproductive tract Cd levels with insulin resistance, dyslipidemia and estrous cycle duration, cystic and atretic follicles, LH levels and inflammation. Thus, these data suggest that subacute exposure to Cd at a dose similar to that of occupational exposure alters the function of the HPG axis, leading to PCOS and FOP features besides metabolic abnormalities in female rats.