Oxidative and Inflammatory Imbalance in Placenta and Kidney of sFlt1-Induced Early-Onset Preeclampsia Rat Model

Preeclampsia (PE) is a pregnancy-specific disorder characterized by the new onset of hypertension plus proteinuria and/or end-organ dysfunction. Here, we investigate the role of the nicotinamide adenine dinucleotide phosphate (NADPH) oxidase system as a major component of reactive oxygen species generation, in a rodent model of early-onset preeclampsia induced by excess sFlt1 (soluble fms-like tyrosine kinase 1). Placenta and kidney samples were obtained from normal pregnant and PE rats to measure the sFlt1/PlGF (placental growth factor) ratio in addition to oxidative stress-related parameters, including the activities and expressions of NADPH oxidase isoforms (NOX1, NOX2, and NOX4), components of nitric oxide (NO) metabolism, and antioxidant enzymes. Peroxisome proliferator-activated receptors (PPARα, PPARγ) and cytokines IL1β, IL3, IL6, IL10, and IL18 were also measured to evaluate the inflammation status in our experimental setting. Excessive O2●− production was found in rats that were treated with sFlt1; interestingly, this alteration appears to be mediated mainly by NOX2 in the placenta and by NOX4 in the kidney. Altered NO metabolism and antioxidant defense systems, together with mitochondrial dysfunction, were observed in this model of PE. Preeclamptic animals also exhibited overexpression of proinflammatory biomarkers as well as increased collagen deposition. Our results highlight the role of NADPH oxidase in mediating oxidative stress and possibly inflammatory processes in the placenta and kidney of an sFlt1-based model of early-onset preeclampsia

Retinoprotective Effect of Wild Olive (Acebuche) Oil-Enriched Diet against Ocular Oxidative Stress Induced by Arterial Hypertension

Oxidative stress plays an important role in the pathogenesis of ocular diseases, including hypertensive eye diseases. The beneficial effects of olive oil on cardiovascular diseases might rely on minor constituents. Currently, very little is known about the chemical composition and/or therapeutic effects of the cultivated olive tree’s counterpart, wild olive (also known in Spain as acebuche—ACE). Here, we aimed to analyze the antioxidant and retinoprotective effects of ACE oil on the eye of hypertensive mice made hypertensive via administration of NG-nitro-L-arginine-methyl-ester (L-NAME), which were subjected to a dietary supplementation with either ACE oil or extra virgin olive oil (EVOO) for comparison purposes. Deep analyses of major and minor compounds present in both oils was accompanied by blood pressure monitoring, morphometric analyses, as well as different determinations of oxidative stress-related parameters in retinal layers. Aside from its antihypertensive effect, an ACE oil-enriched diet reduced NADPH (nicotinamide adenine dinucleotide phosphate) oxidase activity/gene/protein expression (with a major implication of NADPH oxidase (NOX)2 isoform) in the retinas of hypertensive mice. Supplementation with ACE oil in hypertensive animals also improved alterations in nitric oxide bioavailability and in antioxidant enzyme profile. Interestingly, our findings show that the use of ACE oil resulted in better outcomes, compared with reference EVOO, against hypertension-related oxidative retinal damage.Consejería de Economía, Conocimiento, Empresas y Universidad, Junta de Andalucía (2017/440; 2020/275; CTS-584)Ministerio de Ciencia e Innovación, Gobierno de España (Plan Estatal de Investigación Científica y Técnica y de Innovación 2017-2020Programa Estatal de I+D+I PID2019-109002RB-I0

Mechanism of vascular toxicity in rats subjected to treatment with a tyrosine kinase inhibitor

Sunitinib (Su) is a tyrosine kinase inhibitor with antiangiogenic and antineoplastic effects that is recommended therapy for renal cell carcinoma, gastrointestinal stromal tumors, and pancreatic neuroendocrine tumors. Arterial hypertension is one of the adverse effects observed in the treatment with Su. The aim of this work was to deepen our understanding of the underlying mechanisms involved in the development of this side effect. Studies on endothelial function, vascular remodeling and nicotinamide adenine dinucleotide phosphate oxidase (NADPH oxidase) system were carried out in thoracic aortas from rats treated with Su for three weeks. Animals subjected to Su treatment presented with increased blood pressure and reduced endothelium-dependent vasodilation, the latter being reverted by NADPH oxidase blockade. Furthermore, vascular remodeling and stronger Masson trichrome staining, together with enhanced immunofluorescence signal for collagen 1 alpha 1 (Col1ff1), were observed in aortas from treated animals. These results were accompanied by a significant elevation in superoxide anion production and the activity/protein/gene expression of NADPH oxidase isoforms (NOX1, NOX2, and NOX4), which was also prevented by NOX inhibition. Furthermore, a decrease in nitric oxide (NO) levels and endothelial nitric oxide synthase (eNOS) activation was observed in aortas from Su-treated animals. All these results indicate that endothelial dysfunction secondary to changes in vascular remodeling and oxidative stress might be responsible for the typical arterial hypertension that develops following treatment with Su.Junta de Andalucía 2017/44

Anti‐inflammatory action of dietary wild olive (Acebuche) oil in the retina of hypertensive mice

Inflammation plays a crucial role in the course of eye diseases, including many vascular retinopathies. Although olive oil is known to have beneficial effects against inflammatory processes, there is no information available on the anti‐inflammatory potential of the wild olive tree (namely, acebuche (ACE) for the primitive Spanish lineages). Here we investigate the anti‐inflammatory effects of ACE oil in the retina of a mouse model of arterial hypertension, which was experimentally induced by administration of L‐NAME (NG‐nitro‐L‐arginine‐methyl‐ester). The animals were fed supplements of ACE oil or extra virgin olive oil (EVOO, for comparative purposes). Retinal function was assessed by electroretinography (ERG), and different inflammation‐related parameters were measured in the retina and choroid. Besides significant prevention of retinal dysfunction shown in ERG recordings, ACE oil‐enriched diet upregulated the expression of the anti‐inflammatory mark-ers PPARγ, PPARα and IL‐10, while reducing that of major proinflammatory biomarkers, IL‐1β, IL‐ 6, TNF‐α and COX‐2. This is the first report to highlight the anti‐inflammatory properties of an ACE oil‐enriched diet against hypertension‐related retinal damage. Noteworthy, dietary supplementa-tion with ACE oil yielded better results compared to a reference EVOO.Ministerio de Ciencia e Innovación PID2019-109002RB-I00Junta de Andalucía 2020/275, 2021/188, CTS-584Universidad de Sevilla 2020/116

Evaluación del daño vascular tras el tratamiento con Sunitinib: disfunción endotelial y mecanismos patogénicos implicados

Sunitinib (Su) is an orally active multitarget tyrosine kin ase inhibitor with antiangiogenic and antineoplastic effects. Su is the first line choice for metastatic renal cell carcinoma treatment ( and the second line one for gastrointestinal stromal tumours (GISTs) and pancreatic neuroendocrine tumour ( pNET). Th e drug binds to several tyrosine kinase receptors including platelet derived growth factor receptor PDGFR α β; colony stimulating factor 1 receptor, CSF 1R; stem cell factor receptor, KIT; Fms like tyrosine kinase 3 receptor, FLT3; vascula r endothelial growth factor receptor, VEGFR 1 2 3; and glial cell line derived neurotrophic factor receptor, RET. A rterial hypertension is a common feature associated to treatment with Su and other VEGFR inhibitors S everal mechanisms have been suggested to explain this side effect , e. reduced nitric oxide generation, endothelial dysfunction, capillary rarefaction or endothelin 1 ove rexpression . However , the precise underlying pathophysiological mechanism s remain unknown. The aim of this doctoral thesis was to deepen our understanding o n those mechanisms involved in the development of Su depen d ent hypertension . To this purpose , our experi m ental approach focused on the stud y o f vascular function, including assays in the presence or absence of circula ting extracellular vesicles. In addition , vascular remodel l ing, fibrosis and inflammation, nicotinamide adenine dinucleoti de phosphate oxidase (NADPH oxidase), oxidative stress and nitric oxide systems were evaluated in thoracic aortas obtained from male Wistar rats treated with Su for 3 weeks. Our results show ed increased blood pressure and reduced endotheliu m dependent vasodilation in animals subjected to Su treatment , the latter being reverted by NADPH ox idase blockade . Interestingly, reduced endothelium dependent vasodilation was also observed in aortic rings of normotensive rats preincubated with circulating extracellular vesicles from Su treated rats, which was reverted by oxidative blockade . Further alterations observed in aortas from Su treated animals included vascular remode l ling and stronge r Masson trichrome staining, together with enhanced immunofluorescence signal for collagen 1 alpha 1 (Col1α1). These results were accompanied by a significant elevation in superoxide anion production and in the activity/protein/gene expression of NADPH oxidase isoforms (NOX1, NOX2 and NOX4), which could be prevented by NOX inhibition. Su treated animals also presented with NOX dependent eNOS inactivation and decrease d aortic NO levels. All these observations indicate that endothelial dysfunction secondary to enhanced oxidative stress and changes in vascular remodel l ing might be responsible for the establishment of arterial hypertension that typically occurs followi n g treatment with Su

Impact of maternal nutrition in viral infections during pregnancy

Other than being a physiological process, pregnancy is a condition characterized by major adaptations of maternal endocrine and metabolic homeostasis that are necessary to accommodate the fetoplacental unit. Unfortunately, all these systemic, cellular, and molecular changes in maternal physiology also make the mother and the fetus more prone to adverse outcomes, including numerous alterations arising from viral infections. Common infections during pregnancy that have long been recognized as congenitally and perinatally transmissible to newborns include toxoplasmosis, rubella, cytomegalovirus, and herpes simplex viruses (originally coined as ToRCH infections). In addition, enterovirus, parvovirus B19, hepatitis virus, varicella-zoster virus, human immunodeficiency virus, Zika and Dengue virus, and, more recently, coronavirus infections including Middle Eastern respiratory syndrome (MERS) and severe acute respiratory syndrome (SARS) infections (especially the novel SARS-CoV-2 responsible for the ongoing COVID-19 pandemic), constitute relevant targets for current research on maternal-fetal interactions in viral infections during pregnancy. Appropriate maternal education from preconception to the early postnatal period is crucial to promote healthy pregnancies in general and to prevent and/or reduce the impact of viral infections in particular. Specifically, an adequate lifestyle based on proper nutrition plans and feeding interventions, whenever possible, might be crucial to reduce the risk of virus-related gestational diseases and accompanying complications in later life. Here we aim to provide an overview of the emerging literature addressing the impact of nutrition in the context of potentially harmful viral infections during pregnancy.This work was supported by Consejería de Transformación Económica, Industria, Conocimiento y Universidades, Junta de Andalucía, Spain (2020/275; CTS-584); Fondo Nacional de Desarrollo Científico y Tecnológico (FONDECYT, grant number 1190316) and International Sabbatical (University Medical Centre Groningen, University of Groningen, The Netherlands) from the Vice-Rectorate of Academic Affairs, Academic Development Office of the Pontificia Universidad Católica de Chile, Chile. AS is recipient of an FPU predoctoral fellowship from Ministerio de Ciencia, Innovación y Universidades (FPU17/03465). CRG was supported by Ministerio de Ciencia e Innovación, Ayudas para la Promoción de Empleo Joven e Implantaci´on de la Garantía Juvenil en I+D+i 2017-2020 (PEJ2018-004474-A).Peer reviewe

Sunitinib-induced oxidative imbalance and retinotoxic effects in rats

[Aims] Sunitinib (Su), a tyrosine kinase inhibitor, is one of the most commonly used anti-angiogenic drugs. Some studies have described retinal detachment and photoreceptor damage following systemic exposure to Su, despite beneficial effects achieved with local treatment of ocular pathologies. The aim of this study was to explore the role of NADPH oxidase system and oxidative stress in eyes from Su-treated animals.[Main methods] Male Wistar rats were administered 25 mg Su/kg body weight/day incorporated in the chow for 3 weeks. Upon treatment completion, NADPH oxidase activity and ROS levels were measured in ocular tissue by chemiluminescence and dihydroethidium (DHE) staining, respectively. The expression of NADPH oxidase isoforms (NOX1, NOX2 and NOX4), antioxidant enzymes and endothelial/inducible nitric oxidase isoforms (eNOS/iNOS) in the eyecup and/or retina were measured via immunofluorescence, immunoblotting and RT-qPCR.[Key findings] NADPH oxidase activity/expression increased in eyecup and retinas from Su-treated rats. Immunohistofluorescence studies in retinal layer confirmed a higher signal of NADPH oxidase isoforms after Su treatment. Treated animals also presented with reductions in NO levels and eNOS expression, whereas iNOS was upregulated. Finally, a significant depletion of antioxidant enzyme glutathione peroxidase was measured in eyecups of rats following Su exposure, and the opposite pattern was seen for glutathione reductase and superoxide dismutase.[Significance] This study demonstrates that Su treatment is associated with NADPH oxidase-derived oxidative stress in the eye. Long-term treatment of Su should be properly monitored to avoid retinotoxic effects that might result in ocular pathologies and sight-threatening conditions.This study was supported by Consejería de Economía, Conocimiento, Empresas y Universidad, Junta de Andalucía (2017/00000440; CTS-584). AS is recipient of an FPU predoctoral fellowship from Ministerio de Ciencia, Innovación y Universidades (FPU17/03465). CR-G was supported by Ministerio de Ciencia, Innovación y Universidades, Ayudas para la Promoción de Empleo Joven e Implantación de la Garantía Juvenil en I+D+i 2017-2020 (PEJ2018-004474-A)

Hypertension secondary to nitric oxide depletion produces oxidative imbalance and inflammatory/fibrotic outcomes in the cornea of C57BL/6 mice.

Arterial hypertension (AH) leads to oxidative and inflammatory imbalance that contribute to fibrosis development in many target organs. Here, we aimed to highlight the harmful effects of severe AH in the cornea. Our experimental model was established by administration of NG-nitro-L-arginine-methyl-ester (L-NAME) to C57BL/6 mice, which were monitored weekly for arterial blood pressure and intraocular pressure (IOP). Morphological studies of ocular tissues were accompanied by analyses of reactive oxygen species generation, and localization/expression of NAPDH oxidase isoforms (NOX1, NOX2, NOX4) and inflammatory biomarkers (PPARα, PPARγ, IL-1β, IL-6, IL-10, TNF-α, and COX-2). Masson's trichrome and Sirius Red staining were used to explore the fibrotic status of the cornea. The expression of collagen isoforms (COL1α1, COL1α2, COL3α1, COL4α1, COL4α2) and relevant metalloproteinases (MMPs) and their tissue inhibitors (TIMPs) were also quantified to evaluate the participation of collagen metabolism in AH-related corneal damage. Hypertensive animals showed an increase in IOP values, and a thinner cornea compared with normotensive controls. Moreover, AH increased NADPH oxidase activity and reactive oxygen species generation in the cornea, which was accompanied by transcriptional upregulation of NOX isoforms and inflammatory biomarkers, while reducing PPAR expression. L-NAME-treated animals also developed corneal fibrosis with overexpression of collagen isoforms and reduction of factors responsible for collagen degradation. This is the first study reporting structural changes in the cornea and elevated IOP in L-NAME-treated mice. Overexpression of the NADPH oxidase system and collagen deposition might play a substantial role in the pathogenic mechanisms contributing to ocular disturbances in a context of severe hypertension

Mechanism of Vascular Toxicity in Rats Subjected to Treatment with a Tyrosine Kinase Inhibitor

Sunitinib (Su) is a tyrosine kinase inhibitor with antiangiogenic and antineoplastic effects that is recommended therapy for renal cell carcinoma, gastrointestinal stromal tumors, and pancreatic neuroendocrine tumors. Arterial hypertension is one of the adverse effects observed in the treatment with Su. The aim of this work was to deepen our understanding of the underlying mechanisms involved in the development of this side effect. Studies on endothelial function, vascular remodeling and nicotinamide adenine dinucleotide phosphate oxidase (NADPH oxidase) system were carried out in thoracic aortas from rats treated with Su for three weeks. Animals subjected to Su treatment presented with increased blood pressure and reduced endothelium-dependent vasodilation, the latter being reverted by NADPH oxidase blockade. Furthermore, vascular remodeling and stronger Masson trichrome staining, together with enhanced immunofluorescence signal for collagen 1 alpha 1 (Col1α1), were observed in aortas from treated animals. These results were accompanied by a significant elevation in superoxide anion production and the activity/protein/gene expression of NADPH oxidase isoforms (NOX1, NOX2, and NOX4), which was also prevented by NOX inhibition. Furthermore, a decrease in nitric oxide (NO) levels and endothelial nitric oxide synthase (eNOS) activation was observed in aortas from Su-treated animals. All these results indicate that endothelial dysfunction secondary to changes in vascular remodeling and oxidative stress might be responsible for the typical arterial hypertension that develops following treatment with Su.This study was supported by Junta de Andalucía - Consejería de Economía, Conocimiento, Empresas y Universidad (2017/440). We thank Centro de Innovación, Tecnología e Innovación de la Universidad de Sevilla (CITIUS, Servicio de Biología y Microscopía) for technical support. This study was supported by Junta de Andalucía-Consejería de Economía, Conocimiento, Empresas y Universidad (2017/440). C.R.-G. was supported by Ministerio de Ciencia, Innovación y Universidades, Ayudas para la Promoción de Empleo Joven e Implantación de la Garantía Juvenil en I+D+i 2017-2020 (PEJ2018-004474-A). A.S.-G. is the recipient of an FPU predoctoral fellowship from Ministerio de Ciencia, Innovación y Universidades (FPU17/03465)