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Reactive oxygen species (ROS), in particular, H2O2, is essential for full activation of VEGF receptor2 (VEGFR2) signaling involved in endothelial cell (EC) proliferation and migration. Extracellular superoxide dismutase (ecSOD) is a major secreted extracellular enzyme that catalyzes the dismutation of superoxide to H2O2, and anchors to EC surface through heparinbinding domain (HBD). Mice lacking ecSOD show impaired postnatal angiogenesis. However, it is unknown whether ecSODderived H2O2 regulates VEGF signaling. Here we show that gene transfer of ecSOD, but not ecSOD lacking HBD (ecSODDHBD), increases H2O2 levels in adductor muscle of mice, and promotes angiogenesis after hindlimb ischemia. Mice lacking ecSOD show reduction of H2O2 in non-ischemic and ischemic limbs. In vitro, overexpression of ecSOD, but not ecSOD-DHBD, in cultured medium in ECs enhances VEGF-induced tyrosine phosphorylation of VEGFR2 (VEGFR2-pY), which is prevented by short-term pretreatment with catalase that scavenges extracellular H2O2. Either exogenous H2O2 (<500 mM), which is diffusible, or nitric oxide donor has no effect on VEGF-induced VEGFR2-pY. These suggest that ecSOD binding to ECs via HBD is required for localized generation of extracellular H2O2 to regulate VEGFR2-pY. Mechanistically, VEGF-induced VEGFR2-pY in caveolae/lipid rafts, but non-lipid rafts, is enhanced by ecSOD, which localizes at lipid rafts via HBD. One of the targets of ROS is protein tyrosine phosphatases (PTPs). ecSOD induces oxidation and inactivation of both PTP1B and DEP1, which negatively regulates VEGFR2-pY, in caveolae/lipid rafts, but not non-lipid rafts. Disruption of caveolae/lipid rafts, or PTPs inhibitor orthovanadate, or siRNAs for PTP1B and DEP1 enhances VEGF-induced VEGFR2-pY, which prevents ecSODinduced effect. Functionally, ecSOD promotes VEGF-stimulated EC migration and proliferation. In summary, extracellular H2O2 generated by ecSOD localized at caveolae/lipid rafts via HBD promotes VEGFR2 signaling via oxidative inactivation of PTPs in these microdomains. Thus, ecSOD is a potential therapeutic target for angiogenesis-dependent cardiovascular diseases. [ABSTRACT FROM AUTHOR]

The article focuses on the role of Menkes Adenosine triphosphatase (MNK) in hypertension induced by angiotensin II. It states that extracellular superoxide dismutase (SOD3) regulates angiotensin II by modulating SOD3 levels. It mentions that chronic angiotensin II infusion enhances the vascular superoxide anion production and systolic blood pressure in MNK mutant mice. It states that coimmunoprecipitation analysis found that angiotensin II stimulation promoted an association of MNK with SOD3.

Abstract: Objectives: Peroxisome proliferator-activated receptors (PPAR) α and β/δ are essential transcriptional regulators of fatty acid oxidation in the heart. However, little is known about the roles of PPARγ in the heart. The present study is to investigate in vivo role(s) of PPARγ in the heart. Methods: A Cre–loxP mediated cardiomyocyte-restricted PPARγ knockout line was investigated. In these mice, exon 1 and 2 of PPARγ were targeted to eliminate PPARγ from cardiomyocytes. Results: PPARγ null mice exhibited pathological changes around 3 months of age, featuring progressive cardiac hypertrophy with mitochondrial oxidative damage. Most mice died from dilated cardiomyopathy. Cardiac expression of Sod2 (encoding manganese superoxide dismutase; MnSOD), a mitochondrial antioxidant enzyme was downregulated both in transcript and protein levels in cardiac samples in PPARγ knockout mice independent of pathological changes. Promoter analyses revealed that Sod2 is a target gene of PPARγ. Consequently, myocardial superoxide content in PPARγ knockout mice was increased, leading to extensive oxidative damage. Treatment with a SOD mimetic compound, MnTBAP, prevented superoxide-induced cardiac pathological changes in PPARγ knockout mice. Conclusions: The present study demonstrates that PPARγ is critical to myocardial redox homeostasis. These findings should provide new insights into understanding the roles of PPARγ in the heart. [Copyright &y& Elsevier]

Protein levels and polymorphisms of p22phox have been suggested to modulate vascular NAD(P)H oxidase activity and vascular production of reactive oxygen species (ROS). We sought to determine whether increasing p22phox expression would alter vascular ROS production and hemodynamics by targeting p22phox expression to smooth muscle in transgenic (Tg) mice. Aortas of Tgp22smc mice had increased p22phox and Nox 1 protein levels and produced more superoxide anti H2O2. Surprisingly, endothelium-dependent relaxation and blood pressure in Tgp22smc mice were normal. Aortas of Tgp22smc mice produced twofold more nitric oxide (NO) at baseline and sevenfold more NO in response to calcium ionophore as detected by electron spin resonance. Western blot analysis revealed a twofold increase in endothelial NO synthase (eNOS) protein expression in Tgp22smc mice. Both eNOS expression and NO production were normalized by infusion of the glutathione peroxidase mimetic ebselen or by crossing Tgp22smc mice with mice overexpressing catalase. We have previously found that NO stimulares extracellular superoxide dismutase (ecSOD) expression in vascular smooth muscle. In keeping with this, aortic segments from Tgp22smc mice expressed twofold more ecSOD, and chronic treatment with the NOS inhibitor NG-nitro-L-arginine methyl ester normalized this, suggesting that NO regulates ecSOD protein expression in vivo. These data indicate that chronic oxidative stress caused by excessive H2O2 production evokes a compensatory response involving increased eNOS expression and NO production. NO in turn increases ecSOD protein expression and counterbalances increased ROS production leading to the maintenance of normal vascular function and hemodynamics. [ABSTRACT FROM AUTHOR]

Excessive production and/or inadequate removal of reactive oxygen species, especially superoxide anion (O2•−), have been implicated in the pathogenesis of many cardiovascular diseases, including atherosclerosis, hypertension, diabetes, and in endothelial dysfunction by decreasing nitric oxide (NO) bioactivity. Since the vascular levels of O2•− are regulated by the superoxide dismutase (SOD) enzymes, a role of SOD in the cardiovascular disease is of substantial interest. Particularly, a major form of SOD in the vessel wall is the extracellular SOD (ecSOD). This review will discuss the characteristics of ecSOD and the role of ecSOD in cardiovascular diseases. [Copyright &y& Elsevier]

In order to establish a scientific basis for the potential use of type I interferons (IFNs) in clinical uveitis, we examined the effect of a preparation of IFN-alpha/beta on experimental autoimmune uveoretinitis (EAU). Lewis rats were immunized with bovine interphotoreceptor retinoid-binding protein (IRBP) and given daily intramuscular injections of 10(5) IU mouse natural IFN-alpha/beta. Intraocular inflammation was assessed by slit-lamp biomicroscopy and histopathological examination. Rats treated daily with IFN-alpha/beta starting on the day of IRBP immunization showed decreased intraocular inflammation as well as a slight delay in onset of inflammation when compared to control rats. This effect was also observed to a lesser extent in rats treated during either the induction phase of EAU only, or starting immediately after the onset of inflammation in the effector phase of EAU. Measurement of IRBP-stimulated splenocyte proliferation and serum anti-IRBP antibody subtypes did not reveal a significant difference between IFN-alpha/beta-treated rats and control rats. Measurement of cytokine production by IRBP-stimulated splenocytes in vitro showed significantly decreased TNF-alpha for IFN-alpha/beta-treated rats compared to control, but no difference for IFN-gamma, IL-2, IL-4, and IL-10. These results indicate that systemic administration of IFN-alpha/beta suppresses IRBP-induced EAU in rats, and suggest that such suppression may be mediated in part by a reduction in TNF-alpha production. [ABSTRACT FROM AUTHOR]

Objectives: The role of endothelin-1 (ET-1) in the pathogenesis of coronary artery spasm is not well understood. We aimed to determine if ET-1 is involved in serotonin-induced coronary spasm in the swine model. Methods: In 10 miniature pigs, a segment of the left anterior descending coronary artery was denuded and irradiated with X-ray. Three months after endothelial denudation, coronary vasomotion was assessed in vivo by quantitative arteriography. Results: Intracoronary serotonin at 10 μg/kg provoked coronary spasm (augmented narrowing of the luminal diameter) at the denuded site (diameter reduction 93 ± 4%) but not at the non-denuded control site (19 ± 4%, P < 0.01) associated with ST segment elevation in the region perfused by the denuded artery. Intracoronary administration of ET-1 at 25 ng/kg caused mild vasoconstriction of the denuded (26 ± 4) and non-denuded site (16 ± 3%, n.s.), but provoked ST segment elevation in the regions perfused by both the denuded and non-denuded arteries. The treatment with an endothelin antagonist (BQ123 0.1 mg/kg) significantly attenuated coronary vasoconstriction and ST segment elevation evoked with ET-1, but did not alter serotonin-induced vasoconstriction either at the denuded or control site. Conclusions: The results of this study suggest that endogenous ET-1 may not be involved in the pathogenesis of serotonin-induced coronary spasm in our swine model. [ABSTRACT FROM PUBLISHER]

Angiogenesis, a new vessel formation from the pre-existing ones, is essential for embryonic development, wound repair and treatment of ischemic heart and limb diseases. However, dysregulated angiogenesis contributes to various pathologies such as diabetic retinopathy, atherosclerosis and cancer. Reactive oxygen species (ROS) derived from NADPH oxidase (NOX) as well as mitochondria play an important role in promoting the angiogenic switch from quiescent endothelial cells (ECs). However, how highly diffusible ROS produced from different sources and location can communicate with each other to regulate angiogenesis remains unclear. To detect a localized ROS signal in distinct subcellular compartments in real time in situ, compartment-specific genetically encoded redox-sensitive fluorescence biosensors have been developed. Recently, the intercellular communication, "cross-talk", between ROS derived from NOX and mitochondria, termed "ROS-induced ROS release", has been proposed as a mechanism for ROS amplification at distinct subcellular compartments, which are essential for activation of redox signaling. This "ROS-induced ROS release" may represent a feed-forward mechanism of localized ROS production to maintain sustained signaling, which can be targeted under pathological conditions with oxidative stress or enhanced to promote therapeutic angiogenesis. In this review, we summarize the recent knowledge regarding the role of the cross-talk between NOX and mitochondria organizing the sustained ROS signaling involved in VEGF signaling, neovascularization and tissue repair. [ABSTRACT FROM AUTHOR]

Presents a study which determined whether the Menkes ATPase (MNK) is required for the activation of extracellular superoxide dismutase (SOD3) in the vasculature. Expression of MNK in vascular smooth muscle cells, vascular endothelial cells, fibroblasts and aorta; Role of vascular MNK for full activation of SOD3; Colocalization of SOD3 with MNK in the trans-Golgi network; Interaction of SOD3 with MNK.

We report a patient who developed multiple liver abscesses and sepsis caused by lactobacilli after the percutaneous intratumoral injection of ethanol for hepatocellular carcinoma. We diagnosed the liver abscess at an early stage because of a the finding of gas on ultrasound and computed tomography. Blood cultures grew Gram-positive rods, which were of the Lactobacillus species. The patient responded to the administration of antibiotics, and his hepatic tumors have not recurred in the 7 months since treatment. This is the first report of liver abscess following percutaneous ethanol injection therapy. [ABSTRACT FROM AUTHOR]

Myocardial infarction (MI) is the primary cause of cardiovascular mortality, and therapeutic strategies to prevent or mitigate the consequences of MI are a high priority. Cardiac progenitor cells (CPCs) have been used to treat cardiac injury post-MI, and despite poor engraftment, they have been shown to inhibit apoptosis and to promote angiogenesis through poorly understood paracrine effects. We previously reported that the direct injection of exosomes derived from CPCs (CPCexo) into mouse hearts provides protection against apoptosis in a model of acute ischemia/reperfusion injury. Moreover, we and others have reported that reactive oxygen species (ROS) derived from NADPH oxidase (NOX) can enhance angiogenesis in endothelial cells (ECs). Here we examined whether bioengineered CPCexo transfected with a pro-angiogenic miR-322 (CPCexo-322) can improve therapeutic efficacy in a mouse model of MI as compared to CPCexo. Systemic administration of CPCexo-322 in mice after ischemic injury provided greater protection post-MI than control CPCexo, in part, through enhanced angiogenesis in the border zones of infarcted hearts. Mechanistically, the treatment of cultured human ECs with CPCexo-322 resulted in a greater angiogenic response, as determined by increased EC migration and capillary tube formation via increased Nox2-derived ROS. Our study reveals that the engineering of CPCexo via microRNA (miR) programing can enhance angiogenesis, and this may be an effective therapeutic strategy for the treatment of ischemic cardiovascular diseases. [ABSTRACT FROM AUTHOR]

Increased aerobic glycolysis in endothelial cells of atheroprone areas of blood vessels has been hypothesized to drive increased inflammation and lesion burden but direct links remain to be established. Here we show that endothelial cells exposed to disturbed flow in vivo and in vitro exhibit increased levels of protein kinase AMP-activated (PRKA)/AMP-activated protein kinases (AMPKs). Selective deletion of endothelial Prkaa1, coding for protein kinase AMP-activated catalytic subunit alpha1, reduces glycolysis, compromises endothelial cell proliferation, and accelerates the formation of atherosclerotic lesions in hyperlipidemic mice. Rescue of the impaired glycolysis in Prkaa1-deficient endothelial cells through Slc2a1 overexpression enhances endothelial cell viability and integrity of the endothelial cell barrier, and reverses susceptibility to atherosclerosis. In human endothelial cells, PRKAA1 is upregulated by disturbed flow, and silencing PRKAA1 reduces glycolysis and endothelial viability. Collectively, these results suggest that increased glycolysis in the endothelium of atheroprone arteries is a protective mechanism. Increased glycolysis and inflammatory responses have been observed in endothelial cells exposed to disturbed flow. However, the role of endothelial glycolysis in atherosclerosis is unclear. Here the authors unveil a protective role for glycolysis by showing that endothelial deletion of Prkaa1 accelerates atherosclerosis in hyperlipidemic mice through a reduction of glycolytic metabolism. [ABSTRACT FROM AUTHOR]

The article reviews a study on the role of mitochondrial thioredoxin (Trx2) as a novel regulator for nicotine adenine dinucleotide phosphate (NADPH) oxidase, endothelial function and systolic blood pressure in angiotensin II-induced hypertension. It suggests that mitochondrial reactive oxygen species (ROS) increases expression of NADPH oxidase components. It also agrees with the finding about the protective effect of Trx2 and the inhibition of cardiac hypertrophy and cardiac superoxide levels by Trx2.

Because of their ability to generate superoxide, we hypothesized that renal vasoconstrictor responses to norepinephrine (NE) and angiotensin II (Ang II) would be enhanced in mice lacking extracellular superoxide dismutase (ecSOD-KO). NE (30 to 300 µg/kg) and Ang II (0.3 to 6 µg/kg) were administered (i.v.) to anesthetized male and female C57BI/6 (WT) and ecSOD-KO mice on a C57B1/6 background (n=7-10). Blood pressure (BP) and renal blood flow (RBF) responses were measured via carotid artery catheter and ultrasonic flow probe, respectively. In WT mice, the decrease of RBF in response to NE was similar between males and females. RBF responses in male and female ecSOD-KO mice to NE were reduced compared to male and female WT mice (e.g RBF response to 30 µg/kg NE in males: -7.9±1.2 vs -20.7±3.8%, p<0.0001; and in females: -12.2±2.3 vs -15.8±2.9%, p=0.04). However, responses in male ecSOD-KO mice were attenuated to a greater extent than in female ecSOD-KO mice, demonstrated by a lesser response in male versus females ecSOD-KO mice (e.g RBF response to 30 µg/kg: -7.9±1.2 vs - 12.2±2.3%, p=0.03). Similar results were found for BP responses to NE, and for RBF and BP responses to Ang II. Contrary to our hypothesis, renal vasoconstrictor responses in ecSOD-KO mice were reduced. We hypothesize that male ecSOD-KO mice have compensatory mechanisms that completely obviate the potential role of increased superoxide in the renal vasculature. [ABSTRACT FROM AUTHOR]

Inflammation, oxidative stress, and copper (Cu) play an important role in cardiovascular disease, including atherosclerosis. We previously reported that cytosolic Cu chaperone antioxidant-1 (Atox1) translocates to the nucleus in response to inflammatory cytokines or exogenous Cu and that Atox1 is localized at the nucleus in the endothelium of inflamed atherosclerotic aorta. However, the roles of nuclear Atox1 and their function are poorly understood. Here we showed that Atox1 deficiency in ApoE−/− mice with a Western diet exhibited a significant reduction of atherosclerotic lesion formation. In vitro, adenovirus-mediated overexpression of nuclear-targeted Atox1 (Ad-Atox1-NLS) in cultured human endothelial cells (ECs) increased monocyte adhesion and reactive oxygen species (ROS) production compared to control cells (Ad-null). To address the underlying mechanisms, we performed genome-wide mapping of Atox1-regulated targets in ECs, using an unbiased systemic approach integrating sequencing data. Combination of ChIP-Seq and RNA-Seq analyses in ECs transfected with Ad-Atox1-NLS or Ad-null identified 1387 differentially expressed genes (DEG). Motif enrichment assay and KEGG pathway enrichment analysis revealed that 248 differentially expressed genes, including inflammatory and angiogenic genes, were regulated by Atox1-NLS, which was then confirmed by real-time qPCR. Among these genes, functional analysis of inflammatory responses identified CD137, CSF1, and IL5RA as new nuclear Atox1-targeted inflammatory genes, while CD137 is also a key regulator of Atox1-NLS-induced ROS production. These findings uncover new nuclear Atox1 downstream targets involved in inflammation and ROS production and provide insights into the nuclear Atox1 as a potential therapeutic target for the treatment of inflammatory diseases such as atherosclerosis. [ABSTRACT FROM AUTHOR]

Exosomes, key mediators of cell‐cell communication, derived from type 2 diabetes mellitus (T2DM) exhibit detrimental effects. Exercise improves endothelial function in part via the secretion of exosomes into circulation. Extracellular superoxide dismutase (SOD3) is a major secretory copper (Cu) antioxidant enzyme that catalyzes the dismutation of O2•− to H2O2 whose activity requires the Cu transporter ATP7A. However, the role of SOD3 in exercise‐induced angiogenic effects of circulating plasma exosomes on endothelial cells (ECs) in T2DM remains unknown. Here, we show that both SOD3 and ATP7A proteins were present in plasma exosomes in mice, which was significantly increased after two weeks of volunteer wheel exercise. A single bout of exercise in humans also showed a significant increase in SOD3 and ATP7A protein expression in plasma exosomes. Plasma exosomes from T2DM mice significantly reduced angiogenic responses in human ECs or mouse skin wound healing models, which was associated with a decrease in ATP7A, but not SOD3 expression in exosomes. Exercise training in T2DM mice restored the angiogenic effects of T2DM exosomes in ECs by increasing ATP7A in exosomes, which was not observed in exercised T2DM/SOD3−/− mice. Furthermore, exosomes overexpressing SOD3 significantly enhanced angiogenesis in ECs by increasing local H2O2 levels in a heparin‐binding domain‐dependent manner as well as restored defective wound healing and angiogenesis in T2DM or SOD3−/− mice. In conclusion, exercise improves the angiogenic potential of circulating exosomes in T2DM in a SOD3‐dependent manner. Exosomal SOD3 may provide an exercise mimetic therapy that supports neovascularization and wound repair in cardiometabolic disease. [ABSTRACT FROM AUTHOR]

1. Angiogenesis, the formation of new capillaries from existing vasculature, is a critical process in normal physiology as well as several physiopathologies. A desire to curb the supportive role angiogenesis plays in the development and metastasis of cancers has driven exploration into anti-angiogenic strategies as cancer therapeutics. Key to this, angiogenesis additionally displays an exquisite sensitivity to bioavailable copper. Depletion of copper has been shown to inhibit angiogenesis in a wide variety of cancer cell and xenograft systems. Several clinical trials using copper chelation as either an adjuvant or primary therapy have been conducted. Yet, the biological basis for the sensitivity of angiogenesis remains unclear. Numerous molecules important to angiogenesis regulation have been shown to be either directly or indirectly influenced by copper, yet a clear probative answer to the connection remains elusive. 2. Measurements of copper in biological systems have historically relied on techniques that, although demonstrably powerful, provide little or no information as to the spatial distribution of metals in a cellular context. Therefore, several new approaches have been developed to image copper in a biological context. One such approach relies on synchrotron-derived X-rays from third-generation synchrotrons and the technique of high resolution X-ray fluorescence microprobe (XFM) analysis. 3. Recent applications of XFM approaches to the role of copper in regulating angiogenesis have provided unique insight into the connection between copper and cellular behaviour. Using XFM, copper has been shown to be highly spatially regulated, as it is translocated from perinuclear areas of the cell towards the tips of extending filopodia and across the cell membrane into the extracellular space during angiogenic processes. Such findings may explain the heightened sensitivity of this cellular process to this transition metal and set a new paradigm for the kinds of regulatory roles that the spatial dynamics of cellular transition metals may play. [ABSTRACT FROM AUTHOR]

Background: Colorectal cancer remains a deadly cancer due to metastatic disease. To understand the molecular mechanisms of metastasis in colon cancer, we investigated whether the copper chaperone antioxidant-1 (Atox1) protein plays a role in this process. Recent findings indicate that Atox1 protein has transcription factor activities and plays a vital role in cell proliferation in cancer cells. However, the role of Atox1 in metastasis has not been examined. Methods: Atox1 expression was determined by immunofluorescence in a tissue microarray generated from a spectrum of CRC patients. Subcellular fractionation of colon cancer cell lines SW480 and SW620 cells was used to examine the cellular location of Atox1 in the face of activin A, a cytokine that stimulates colon cancer metastasis. Atox1 expression was genetically manipulated and cellular migration measured through trans-well assay and proliferation measured by colony formation assays. Results: Here we demonstrate that in patients with metastatic colon cancer, there is a significant increase in the expression of nuclear Atox1. Interestingly, the metastatic CRC cell line SW620 has increased nuclear localization of Atox1 compared to its related non-metastatic cell line SW480. Further, inhibition of endogenous Atox1 by siRNA in SW620 decreased colony formation and reactive oxygen species generation via decreased expression of Atox1 targets cyclin D1 and NADPH oxidase subunit p47 phox, respectively. Additionally, overexpression of nuclear-targeted but not copper binding domain-mutated Atox1 in SW480 cells increased colony formation and cell migration that was further augmented by activin A stimulation, a known enhancer of colon cancer metastasis. Conclusions: Our findings suggest that nuclear Atox1 might be a new therapeutic target as well as a new biomarker for metastatic colorectal cancer. [ABSTRACT FROM AUTHOR]

R2666 --> 849.6 --> Diabetes, whether insulin‐dependent or independent is a major cause of endothelial dysfunction and a leading risk factor for cardiovascular disease. While compelling evidence indicates that endothelial dysfunction is a precursor and contributor to cardiovascular disease, the etiology of diabetes‐induced endothelial dysfunction remains ill‐defined. Recently alteration in endothelial cell bioenergetics has emerged as a new contributor to vascular diseases. However, whether alterations in endothelial glycolysis, the main endothelial cell bioenergetic pathway contributes to endothelial dysfunction is unknown. Herein, we tested the hypothesis that diabetes‐associated endothelial dysfunction involved increases in endothelial cells glycolysis and upregulation of its primary regulator, 6‐phosphofructo‐2‐kinase/fructose‐2, 6‐bisphosphatase 3 (PFKFB3). Measurement of glycolytic activity via Seahorse analyzer in primary human aortic endothelial cells (HAEC) from control and diabetic patients revealed a significant increase in basal glycolysis in diabetic subjects (p<0.05). We also employed Akita mice, a mouse model of type 1 diabetes. Male Akita mice showed a diabetic phenotype with 3‐folds increase in blood glucose. Seahorse analysis in intact aortic explants revealed a 1.7‐fold increase in basal glycolysis and a 1.3‐fold increase in maximal glycolysis in Akita mice (P<0.05). The increase in glycolysis was endothelial‐derived as aortic denudation significantly decreased glycolysis in Akita mice and abolished the difference between groups. No significant difference in mitochondrial function was depicted in the mito‐stress test between Akita and WT aortic explants (p>0.05). Additionally, endothelial cells extracted from aortas of Akita mice exhibited a 2‐fold increase in PFKFB3 mRNA expression(P<0.05). Interestingly, inhibition of PFKFB3 using 3PO (20μM) restored endothelial function in Akita mice. Similarly, Non‐obese diabetic females (NOD), a model for type‐1 diabetes, showed 4‐fold increases in PFKFB3 mRNA expression in aortic endothelial cells and impaired EDR which was restored with 3PO (p<0.05). To further evaluate the role of PFKFB3 in vascular endothelial function we overexpressed PFKFB3 in WT aortas with adenovirus (Ad‐PFKFB3), and reported impaired aorta EDR(P<0.05). Transduction of HAEC with Ad‐PFKFB3 showed a significant increase in ROS producing enzyme (NOX1). Also, inhibition of NOX1 using the selective inhibitor GKT771 restored EDR in Akita and Ad‐PFKFB3‐transduced WT aortas. In Conclusion, our data identified for the first time a role for endothelial glycolysis in the control of vascular relaxation and showed a crucial role of PFKFB3‐mediated endothelial glycolysis in vascular endothelial dysfunction associated with Type‐1 diabetes in males and females. The underlying mechanism potentially involves NOX1 as a downstream target for PFKFB3. Thus, Inhibitors of PFKFB3 could be a potential therapeutic target for diabetes‐induced vascular endothelial dysfunction. [ABSTRACT FROM AUTHOR]