Elisa Pagnin | Università degli Studi di Padova (original) (raw)

Papers by Elisa Pagnin

Journal of Clinical Medicine, 2020

Evidence on cellular/molecular mechanisms leading to atrial fibrillation (AF) are scanty. Increas... more Evidence on cellular/molecular mechanisms leading to atrial fibrillation (AF) are scanty. Increased expression of Rho kinase (ROCK) and myosin-phosphatase-target subunit-1 (MYPT-1), ROCK activity’s marker, were shown in AF patients, which correlated with connexin 40 (Cx40) expression, membrane protein of heart gap junctions, key for rapid action potential’s cell–cell transfer. AF is the most frequent arrhythmia in dialysis patients who present increased MYPT-1 phosphorylation, which correlates with left ventricular (LV) mass. Given ROCK’s established role in cardiovascular–renal remodeling, induction of impaired cell-to-cell coupling/potential conduction promoting AF initiation/perpetuation, we evaluated in dialysis patients with AF, MYPT-1 phosphorylation, Cx40 expression, and their relationships to support their involvement in AF. Mononuclear cells’ MYPT-1 phosphorylation, Cx40 expression, and the ROCK inhibitor fasudil’s effect were assessed in dialysis patients with AF (DPAFs), ...

Journal of Clinical Medicine, 2018

Fabry disease is an X-linked lysosomal storage disease caused by mutations in the GLA gene that l... more Fabry disease is an X-linked lysosomal storage disease caused by mutations in the GLA gene that lead to a reduction or an absence of the enzyme α-galactosidase A, resulting in the progressive and multisystemic accumulation of globotriaosylceramide. Clinical manifestation varies from mild to severe, depending on the phenotype. The main clinical manifestations are cutaneous (angiokeratomas), neurological (acroparesthesias), gastrointestinal (nausea, diarrhea abdominal pain), renal (proteinuria and kidney failure), cardiovascular (cardiomyopathy and arrhythmias), and cerebrovascular (stroke). A diagnosis of Fabry disease can be made with an enzymatic assay showing absent or reduced α-galactosidase A in male patients, while in heterozygous female patients, molecular genetic testing is needed. Enzyme replacement therapy (ERT) with recombinant human α-galactosidase is nowadays the most-used disease-specific therapeutic option. Despite ERT, cardiocerebrovascular-renal irreversible organ in...

PLOS ONE, 2018

Background Fabry disease is characterized by deficient expression/activity of α-GalA with consequ... more Background Fabry disease is characterized by deficient expression/activity of α-GalA with consequent lysosomal accumulation in various organs of its substrate Gb3. Despite enzyme replacement therapy, Fabry disease progresses with serious myocardial, cerebral and renal manifestations. Gb3 accumulation may induce oxidative stress (OxSt), production of inflammatory cytokines and reduction of nitric oxide, which may impact on Fabry disease's clinical manifestations. Methods OxSt status was characterized in 10 patients compared with 10 healthy subjects via protein expression of p22 phox , subunit of NADH/NADPH oxidase, (Western blot), Heme oxygenase (HO)-1 levels (ELISA), antioxidant/anti-inflammatory, lipid peroxidation as malondialdehyde (MDA) production (colorimetric assay), phosphorylation state of Extracellular Signal Regulated Kinase (ERK)1/2 and Myosin Phosphatase Target Protein (MYPT)-1 (Western blot), marker of Rho kinase activation, both involved in OxSt signaling. Cardiac left ventricular (LV) mass was also evaluated (M-mode echocardiography). Results LV mass was higher in Fabry's males (123.72±2.03SEM g/m 2) and females (132.09±6.72g/ m 2). p22 phox expression was also higher in patients (1.04±0.09 d.u. vs 0.54±0.05 d.u. p<0.01) as well as MDA levels (54.51±3.97 vs 30.05±7.11 nmol/mL p = 0.01) while HO-1 was reduced (8.84±0.79 vs 14.03±1.23 ng/mL, p<0.02). MYPT-1's phosphorylation was increased in patients (0.52±0.11 d.u. vs 0.03±0.08 d.u., p<0.01) while phosphorylation of ERK1/2 was reduced (0.91±0.08 d.u. vs 1.53±0.17 d.u., p = 0.004).

Life sciences, Jan 30, 2018

Chronic kidney disease patient's progression to end-stage renal disease as well as their high... more Chronic kidney disease patient's progression to end-stage renal disease as well as their high mortality are linked to cardiovascular disease. However, the high incidence rate of cardiovascular morbidity and mortality in these patients is not fully accounted for by traditional cardiovascular risk factors such as diabetes, hypertension and obesity. Renal disease and CVD are associated with endothelial dysfunction, inflammation and oxidative stress and in this review we will examine what is known regarding their similar roles in both CVD and chronic kidney disease, specifically focusing on the interconnections between oxidative stress, inflammation and endothelial dysfunction. These interconnections are best visualized as a vicious circle wherein these entities coexist and communicate with each other, thereby exacerbating the processes underpinning these different entities with the end result of the high morbidity and mortality that characterize CKD patients. By exploring this vici...

Journal of the American Heart Association, Jan 21, 2016

Tubulointerstitial fibrosis, the final outcome of most kidney diseases, involves activation of ep... more Tubulointerstitial fibrosis, the final outcome of most kidney diseases, involves activation of epithelial mesenchymal transition (EMT). Endothelin-1 (ET-1) activates EMT in cancer cells, but it is not known whether it drives EMT in the kidney. We therefore tested the hypothesis that tubulointerstitial fibrosis involves EMT driven by ET-1. Transgenic TG[mRen2]27 (TGRen2) rats developing fulminant angiotensin II-dependent hypertension with prominent cardiovascular and renal damage were submitted to drug treatments targeted to ET-1 and/or angiotensin II receptor or left untreated (controls). Expressional changes of E-cadherin and α-smooth muscle actin (αSMA) were examined as markers of renal EMT. In human kidney HK-2 proximal tubular cells expressing the ETB receptor subtype, the effects of ET-1 with or without ET-1 antagonists were also investigated. The occurrence of renal fibrosis was associated with EMT in control TGRen2 rats, as evidenced by decreased E-cadherin and increased αSMA...

International Journal of Cardiology, 2006

Antioxid Redox Signal, 2005

Lipid oxidation and environmental pollutants are major sources of alpha,beta-unsaturated aldehyde... more Lipid oxidation and environmental pollutants are major sources of alpha,beta-unsaturated aldehydes such as acrolein and 4-hydroxynonenal. Acrolein (2-propenal), a major product of organic combustion such as tobacco smoke, represents the most reactive alpha,beta-unsaturated aldehyde, with high reactivity toward nucleophilic targets such as sulfhydryl groups. To investigate how acrolein affects respiratory tract cell activation, we exposed either primary (NHBE) or immortalized human bronchial epithelial cells (HBE1) to 0-25 microM acrolein, and determined effects on basal and tumor necrosis factor-alpha (TNFalpha)-induced production of the chemokine interleukin (IL)-8. Cell exposure to acrolein dose-dependently suppressed IL-8 mRNA levels in HBE1 cells (26, 40, and 79% at 5, 10, and 25 microM acrolein concentrations, respectively) and resulted in corresponding decreases in IL-8 production. Studies of nuclear factor-kappaB (NFkappaB) activation, an essential event in IL-8 production, showed decreased TNFalpha-induced NFkappaB activation by acrolein, illustrated by inhibition of nuclear translocation of NFkappaB and reduced IkappaBalpha degradation. Immunochemical analysis of IkappaB kinase (IKK), a redox-sensitive regulator of NFkappaB activation, indicated direct modification of the IKK beta-subunit by acrolein, suggesting that acrolein may act directly on IKK. In summary, our results demonstrate that acrolein can suppress inflammatory processes in the airways by inhibiting epithelial IL-8 production through direct or indirect inhibitory effects on NFkappaB activation.

International Journal of Cardiology, 2016

Life Sciences, 2016

Cardiovascular disease (CVD) is the leading cause of excess mortality in chronic kidney disease (... more Cardiovascular disease (CVD) is the leading cause of excess mortality in chronic kidney disease (CKD) and dialysis patients (DP) who have higher prevalence of left ventricular hypertrophy (LVH), the strongest predictor of CV events. Rho kinase (ROCK) activation is linked in hypertensive patients to cardiac remodeling while ROCK inhibition suppresses cardiomyocyte hypertrophy and, in a human clinical condition opposite to hypertension, its downregulation associates with lack of CV remodeling. Information on ROCK activation-LVH link in CKD and DP is lacking. Materials and methods: Mononuclear cells (PBMCs) MYPT-1 phosphorylation, a marker of ROCK activity, and the effect of fasudil, a ROCK inhibitor, on MYPT-1 phosphorylation were assessed in 23 DPs, 13 stage 3-4 CKD and 36 healthy subjects (HS) by Western blot. LV mass was assessed by M-mode echocardiography. Key findings: DP and CKD had higher MYPT-1 phosphorylation compared to HS (p b 0.001 and p = 0.003). Fasudil (500 and 1000 μM) dose dependently reduced MYPT-1 phosphorylation in DP (p b 0.01). DP had higher LV mass than CKD (p b 0.001). MYPT-1 phosphorylation was higher in patients with LVH (p = 0.009) and correlated with LV mass both in DP and CKD with LVH (p b 0.001 and p = 0.006). Significance: In DP and CKD, ROCK activity tracks with LVH. This ROCK activation-LVH link provided in these CVD high-risk patients along with similar findings in hypertensive patients and added to opposite findings in a human model opposite to hypertension and in type 2 diabetic patients, identify ROCK activation as a potential LVH marker and provide further rationale for ROCK activation inhibition as target of therapy in CVD high-risk patients.

International journal of clinical and experimental medicine, 2015

Recent evidence showed that endogenous nicotinamide adenine dinucleotide phosphate-oxidase 4 (NOX... more Recent evidence showed that endogenous nicotinamide adenine dinucleotide phosphate-oxidase 4 (NOX4) may exert a protective role on the cardiovascular system inducing vasodilation, reduction of blood pressure, and anti-proliferative actions. However, the functional significance of NOX4 in the cardiovascular system in humans remains elusive. Mononuclear cell levels of NOX4 were assessed by immunoblotting in 14 Gitelman's patients (GS), a unique human model of endogenous Ang II signaling antagonism and activation of anti-atherosclerotic and anti-remodeling defenses, and compared to 11 untreated essential hypertensive patients as well as to 11 healthy normotensive subjects. The association between NOX4 and its effector heme oxygenase (HO-1) (sandwich immunoassay) was also evaluated. NOX4 protein levels were decreased in hypertensive patients as compared to both GS and healthy subjects (1.06±0.31 AU vs. 1.76±0.54, P=0.002 and vs. 1.61±0.54, P=0.018, respectively). NOX4 protein level ...

American Journal of Hypertension, 2000

BS and GS are characterized by an abnormal cell signaling (reduced stimulated [Ca ϩϩ ] i and [IP3... more BS and GS are characterized by an abnormal cell signaling (reduced stimulated [Ca ϩϩ ] i and [IP3] i), pointing toward a reduced PKC and cell reactivity. NO system is up regulated (increased ecNOS mRNA, NO2 Ϫ /NO3 Ϫ and cGMP excretion) which further contribute to the vascular hyporeactivity. Since PKC regulates many signaling processes and ecNOS gene expression, we evaluated basal and fMLP-stimulated (10 min, 300 nM) PKC activity PKC␣ translocation rate (Western blot) in the cytosol and membrane of neutrophyls from 3 BS, 6 GS and 10 controls (C). We also studied the effect of PKC stimulation and inhibition on monocyte ecNOS gene expression. Finally, we evaluated monocyte G␣q gene expression, since Gq protein transduces signals to PLC␤, to generate IP3. Cytosol and membrane basal PKC activity were similar in BS-GS and C (70Ϯ3 vs 80Ϯ2; 37Ϯ3 vs 46Ϯ2 pmol/min/mg prot respectively), while fMLP-stimulated membrane PKC activity increased to a lower extent in BS-GS (from 43Ϯ2 to 53Ϯ3 vs 38Ϯ2 to 66Ϯ3 pmol/min/mg prot, pϽ0.05). Membrane PKC␣ expression was similar (8.5Ϯ1.5 vs 12.4Ϯ4.0 d.u.), while its ⌬ increase after fMLP was reduced in BS-GS vs C (4.5Ϯ1.4 vs 9.5Ϯ2.1 pϽ0.01). PMA (100 nM) reduced ecNOS gene expression in BS-GS (from 0.80Ϯ0.05 to 0.55Ϯ0.07 d.u. pϽ0.001), while incubation of control cells with GF109203X, (a PKC inhibitor, 1.2 mM), increased ecNOS mRNA (from 0.41Ϯ0.04 to 0.62Ϯ0.05, d.u. pϽ0.001). G␣q mRNA was reduced in BS-GS vs C (0.87Ϯ0.013 vs 0.98Ϯ0.005 d.u., pϽ0.0004). Our data establish that in BS-GS G␣q is reduced and this may cause the altered cell signaling responsible of reduced vascular contractility through reduced PKC. Reduced PKC-dependent NO system up regulation may further contribute to BS and GS vascular hyporeactivity.

High Blood Pressure & Cardiovascular Prevention, 2015

Insights into the Angiotensin II (Ang II) signalling pathways have been provided by extensive stu... more Insights into the Angiotensin II (Ang II) signalling pathways have been provided by extensive studies using Bartter's/Gitelman's syndromes patients. These syndromes are characterized by activation of the reninangiotensin-aldosterone system but do not develop hypertension and cardiovascular remodelling, therefore represent a mirror image of hypertension and clinically manifest themselves as the opposite of hypertension. The short and the long-term signalling of Ang II remain an important matter of investigation to shed light on mechanisms responsible for the pathophysiology of hypertension and its long-term complications, such as cardiovascular remodelling and atherogenesis. In particular the long-term signalling of Ang II is involved in the pathophysiology of cardiovascular-renal remodelling, inflammatory and hypertrophic responses in which the relationship between RhoA/Rho kinase pathway and NO system plays a crucial role. This review reports the results of our studies in Bartter's and Gitelman's syndromes to get better insight these processes and the role of Ang II signaling. The information obtained from the studies in Bartter's/Gitelman's patients can, in fact, clarify, confirm or be used to gather more general data on the biochemical mechanisms responsible for the pathophysiology of hypertension and its long-term complications and could contribute to identify additional potential significant targets of therapy.

High Blood Pressure & Cardiovascular Prevention, 2005

European Urology Supplements, 2004

International Journal of Molecular Medicine, 2006

Aldosterone seems to play a role in the regulation of the electrolyte content of sperm and in the... more Aldosterone seems to play a role in the regulation of the electrolyte content of sperm and in the motility of spermatozoa. The aim of the study was to evaluate the presence of the mineralocorticoid receptor (MR) in human ejaculated spermatozoa. We have assayed MR on spermatozoa of freshly ejaculated sperm from healthy donors. The identification of MR was made by using immunohistochemistry and immunofluorescence analyses, while MR mRNA expression was evaluated by real-time PCR assay. The immunohistochemical and immunofluorescence analyses showed positive staining both in the midpiece and in the tail of the spermatozoa. Relative quantification of MR by using real-time PCR shows that the mRNA expression of MR in spermatozoa is lower than in mononuclear leukocytes (positive controls). Sequencing showed complete identity between the sequence obtained from spermatozoa and the human MR cDNA sequence. Further studies should be performed in order to elucidate a possible physiological role of aldosterone in regulating electrolyte concentration, and the pro-oxidant effect of excess aldosterone in this new target tissue.

Journal of Biomedical Science, 2011

Background: Angiotensin II (Ang II) signaling occurs via two major receptors which activate non-r... more Background: Angiotensin II (Ang II) signaling occurs via two major receptors which activate non-receptor tyrosin kinases that then interact with protein tyrosin-phosphatases (PTPs) to regulate cell function. SHP-2 is one such important PTP that also functions as an adaptor to promote downstream signaling pathway. Its role in Ang II signaling remains to be clarified. Results: Using cultured normal human fibroblasts, immunoprecipitation and western blots, we show for the first time that SHP-2 and PLCβ1 are present as a preformed complex. Complex PLCβ1 is tyr-phosphorylated basally and Ang II increased SHP-2-PLCβ1 complexes and caused complex associated PLCβ1 tyr-phosphorylation to decline while complex associated SHP-2's tyr-phosphorylation increased and did so via the Ang II type 1 receptors as shown by Ang II type 1 receptor blocker losartan's effects. Moreover, Ang II induced both increased complex phosphatase activity and decreased complex associated PLCβ1 tyr-phosphorylation, the latter response required regulator of G protein signaling (RGS)-2. Conclusions: Ang II signals are shown for the first time to involve a preformed SHP-2-PLCβ1 complex. Changes in the complex's PLCβ1 tyr-phosphorylation and SHP-2's tyr-phosphorylation as well as SHP-2-PLCβ1 complex formation are the result of Ang II type 1 receptor activation with changes in complex associated PLCβ1 tyrphosphorylation requiring RGS-2. These findings might significantly expand the number and complexity of Ang II signaling pathways. Further studies are needed to delineate the role/s of this complex in the Ang II signaling system.

Biochemical and Biophysical Research Communications, 2003

Ozone (O 3) is among the most reactive environmental oxidant pollutants to which cutaneous tissue... more Ozone (O 3) is among the most reactive environmental oxidant pollutants to which cutaneous tissues are exposed. O 3 exposure has been shown to induce antioxidant depletion as well as the oxidation of lipids and proteins within the outermost skin layer, the stratum corneum. However, relatively little is known regarding the potential effects of O 3 on the cellular constituents of the underlying skin epidermis and dermis. In the present study, hairless mice exposed for 6 h to 0.8 ppm O 3 showed increases in lipid peroxidation, as quantitated by increases in 4-hydroxynonenal-protein adducts. O 3 exposure caused an induction of the stress proteins HSP27 and heme oxygenase-1 (HO-1), starting at 6 h and increasing up to 18 h after O 3 exposure. This was accompanied by an increase in matrix metalloproteinase-9 (MMP-9) mRNA and activity levels, indicative of possible injurious-reparative processes. Collectively, our data demonstrate that skin exposure to O 3 not only affects antioxidant levels and oxidation markers in the outermost stratum corneum layer, but also induces cellular stress responses in the deeper cellular layers of the skin.

Cardiorenal Medicine, 2015

Hypothesis/Introduction: Angiotensin II (Ang II) has been shown to control erythropoietin (EPO) s... more Hypothesis/Introduction: Angiotensin II (Ang II) has been shown to control erythropoietin (EPO) synthesis as Ang II type 1 receptor (AT1R) blockers block Ang-II-induced EPO oversecretion. To further explore the involvement of AT1R in processes controlling EPO levels, plasma EPO and mononuclear cell NADPH oxidase 4 (NOX4) - a NOX family member involved in oxygen sensing, which is a process central to controlling EPO levels - were assessed in Bartter's/Gitelman's syndrome (BS/GS) patients, a human model of endogenous AT1R antagonism and healthy subjects. Heme oxygenase (HO)-1, antioxidant and anti-inflammatory factor related to NOX4 activation, and the relationship of EPO and NOX4 to HO-1 were also assessed. Materials and Methods: EPO was measured by chemiluminescent immunoassay, HO-1 by sandwich immunoassay and NOX4 protein expression by Western blot. Results: EPO was increased in BS/GS patients compared to healthy subjects (7.64 ± 2.47 vs. 5.23 ± 1.07 U/l; p = 0.025), wherea...