Co-expression and modulation of neuronal and endothelial nitric oxide synthase in human endothelial cells

Bachetti, Tiziana; Comini, Laura; Curello, Salvatore; Bastianon, D.; Palmieri, Michela; Bresciani, Giuliana; Callea, Francesco; Ferrari, Roberto

doi:10.1016/j.yjmcc.2004.07.006

Cited by 80 publications

(61 citation statements)

References 28 publications

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“…We found that all three NOS isoforms were expressed in HUVECs (Fig. 4A), similar to previously published data from several different groups (4,9,16). Interestingly, nNOS demonstrated a prominent nuclear localization, similar to the findings by several other groups (43,48), the significance of which in endothelial cells (such as on the regulation of protein expression) is currently being examined in our laboratory.…”

Section: E 2 Attenuates High Glucose-induced Nitrotyrosine and Peroxysupporting

confidence: 91%

“…Traditionally, eNOS has been described as the critical enzyme for endothelial NO production although endothelial cells express both eNOS and nNOS even under resting conditions (4,17). However, even a short period (1 h or more) of exposure to high glucose conditions can lead to ROS-mediated uncoupling of eNOS, whereby eNOS yields superoxide instead of NO (20).…”

Section: Discussionmentioning

confidence: 99%

“…Three distinct isoforms of NOS are present in the body, namely, endothelial NOS (eNOS), inducible NOS (iNOS), and neuronal NOS (nNOS) (34). Classically, eNOS has been considered the main source of NO in the vasculature, although there is increasing evidence that nNOS in the endothelium plays an important function (4,9,10,17). Recently, we have demonstrated the presence of endothelial nNOS in rat mesenteric arteries where this enzyme contributes to a vasorelaxant role (33).…”

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confidence: 99%

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Estradiol attenuates high glucose-induced endothelial nitrotyrosine: role for neuronal nitric oxide synthase

Chakrabarti

Cheung

Davidge

2012

American Journal of Physiology-Cell Physiology

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Chakrabarti S, Cheung CC, Davidge ST. Estradiol attenuates high glucose-induced endothelial nitrotyrosine: role for neuronal nitric oxide synthase. Am J Physiol Cell Physiol 302: C666 -C675, 2012. First published November 30, 2011 doi:10.1152/ajpcell.00181.2011.-Hyperglycemia in diabetes causes increased oxidative stress in the vascular endothelium with generation of free radicals such as superoxide. Peroxynitrite, a highly reactive species generated from superoxide and nitric oxide (NO), induces proinflammatory tyrosine nitration of intracellular proteins under such conditions. The female sex hormone estrogen appears to exert protective effects on the nondiabetic endothelium. However, several studies show reduced vascular protection in women with diabetes, suggesting alterations in estrogen signaling under high glucose. In this study, we examined the endothelial effects of estrogen under increasing glucose levels, focusing on nitrotyrosine and peroxynitrite. Human umbilical vein endothelial cells were incubated with normal (5.5 mM) or high (15.5 or 30.5 mM) glucose before addition of estradiol (E 2, 1 or 10 nM). Selective NO synthase (NOS) inhibitors were used to determine the role of specific NOS isoforms. Addition of E2 significantly reduced high glucoseinduced increase in peroxynitrite and consequently, nitrotyrosine. The superoxide levels were unchanged, suggesting effects on NO generation. Inhibition of neuronal NOS (nNOS) reduced high glucoseinduced nitrotyrosine, demonstrating a critical role for this enzyme. E2 increased nNOS activity under normal glucose while decreasing it under high glucose as determined by its phosphorylation status. These data show that nNOS contributes to endothelial peroxynitrite and subsequent nitrotyrosine generation under high glucose, which can be attenuated by E2 through nNOS inhibition. The altered regulation of nNOS by E2 under high glucose is a potential therapeutic target in women with diabetes.

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Section: E 2 Attenuates High Glucose-induced Nitrotyrosine and Peroxysupporting

confidence: 91%

Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

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Estradiol attenuates high glucose-induced endothelial nitrotyrosine: role for neuronal nitric oxide synthase

Chakrabarti

Cheung

Davidge

2012

American Journal of Physiology-Cell Physiology

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“…A possibility to explain this discrepancy is the concomitant raise in the inhibitory phosphorylation of eNOS on Thr495 as previously described (Fleming et al, 2001). Also estradiol could be affecting other sources for NO as is the case of neuronal NOS isoform, described as a functional NO source in HUVEC (Bachetti et al, 2004).…”

Section: Discussionmentioning

confidence: 85%

Estradiol, acting through estrogen receptor alpha, restores dimethylarginine dimethylaminohydrolase activity and nitric oxide production in oxLDL-treated human arterial endothelial cells

Novella

Laguna-Fernández

Lázaro-Franco

et al. 2013

Molecular and Cellular Endocrinology

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a b s t r a c tAsymmetric dimethylarginine (ADMA) is an endogenous inhibitor of nitric oxide (NO) synthase. ADMA accumulation, mainly due to a decreased dimethylarginine dimethylaminohydrolase (DDAH) activity, has been related to the development of cardiovascular diseases. We investigate whether estradiol prevents the changes induced by oxidized low density lipoprotein (oxLDL) on the DDAH/ADMA/NO pathway in human umbilical artery endothelial cells (HUAEC). HUAEC were exposed to estradiol, native LDL (nLDL), oxLDL and their combinations for 24 h. In some experiments, cells were also exposed to the unspecific estrogen receptor (ER) antagonist ICI 182780, the specific ERa antagonist MPP or specific agonists for ERa, ERb and GPER. ADMA concentration was measured by HPLC and concentration of NO by amperometry. Protein expression and DDAH activity were measured by immunoblotting and an enzymatic method, respectively. oxLDL, but not nLDL, increased ADMA concentration with a concomitant decrease on DDAH activity. oxLDL reduced eNOS protein and NO production. Estradiol alone had no effects on DDAH/ADMA/NO pathway, but increased the attenuated endothelial NO production induced by oxLDL by reduction in ADMA and preventing loss of eNOS protein levels. ICI 182780 and MPP completely abolished these effects of estradiol on oxLDL-exposed cells. ERa agonist, but not ERb and GPER agonists, mirrored estradiol effects on NO production. In conclusion, estradiol restores (1) DDAH activity, and therefore ADMA levels, and (2) NO production impaired by oxLDL in HUAEC acting through ERa.

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“…This may perhaps be explained by the citrulline-arginine cycle, reflecting the ability of many nonhepatic cells to convert citrulline to arginine. 36 ECs may also express the neuronal isoform of NOS, 37 although its role in endothelial NO production remains unclear and, accordingly, we did not test the effects of CSE on the neuronal isoform of NOS per se.…”

Section: Discussionmentioning

confidence: 99%

Adverse Effects of Cigarette Smoke on NO Bioavailability

et al. 2006

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Abstract-Endothelial dysfunction is a hallmark of cardiovascular disease, and the L-arginine:NO pathway plays a critical role in determining endothelial function. Recent studies suggest that smoking, a well-recognized risk factor for vascular disease, may interfere with L-arginine and NO metabolism; however, this remains poorly characterized. Accordingly, we performed a series of complementary in vivo and in vitro studies to elucidate the mechanism by which cigarette smoke adversely affects endothelial function. In current smokers, plasma levels of asymmetrical dimethyl-arginine (ADMA) were 80% higher (Pϭ0.01) than nonsmokers, whereas citrulline (17%; PϽ0.05) and N-hydroxy-L-arginine (34%; PϽ0.05) were significantly lower. Exposure to 10% cigarette smoke extract (CSE) significantly affected endothelial arginine metabolism with reductions in the intracellular content of citrulline (81%), N-hydroxy-L-arginine (57%), and arginine (23%), while increasing ADMA (129%). CSE significantly inhibited (38%) arginine uptake in conjunction with a 34% reduction in expression of the arginine transporter, CAT1. In conjunction with these studies, CSE significantly reduced the activity of eNOS and NO production by endothelial cells, while stimulating the production of reactive oxygen species. In conclusion, cigarette smoke adversely affects the endothelial L-arginine NO synthase pathway, resulting in reducing NO production and elevated oxidative stress. In conjunction, exposure to cigarette smoke increases ADMA concentration, the latter being a risk factor for cardiovascular disease. Key Words: smoking Ⅲ endothelium Ⅲ metabolism A therosclerotic coronary and cerebrovascular disease are leading causes of death and disability in the Western world, and cigarette smoking has been clearly identified as a risk factor for coronary artery disease (reviewed by Ambrose and Barua 1 ). In this context, measures of endothelial function have been associated with cardiovascular outcome, 2 and smoking has been widely associated with reduced endothelial function. 3 The endothelium plays a central role in the modulation of vascular tone, the inhibition of platelet aggregation and vascular smooth muscle proliferation, and a key participation in angiogenesis under appropriate conditions. NO is well recognized as playing a pivotal part in these endothelial properties, being produced by the endothelial isoform of NO from its preferred substrate L-arginine. In this context, provision of sufficient L-arginine is critical for the sustained production of NO supply, 4 mediated in endothelial cells (ECs) principally by the type 1 cationic amino acid transporter (CAT1). Previous studies indicate that deleterious actions of cigarette smoke on endothelial function could be mitigated by supplemental L-arginine; 5 however, to date, the precise basis for this interaction remains unclear. One explanation is that cigarette smoke exerts an inhibitory effect on components of the L-arginine:NO pathway to influence NO production. In addition, potential effects of cigar...

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Co-expression and modulation of neuronal and endothelial nitric oxide synthase in human endothelial cells

Cited by 80 publications

References 28 publications

Estradiol attenuates high glucose-induced endothelial nitrotyrosine: role for neuronal nitric oxide synthase

Estradiol attenuates high glucose-induced endothelial nitrotyrosine: role for neuronal nitric oxide synthase

Estradiol, acting through estrogen receptor alpha, restores dimethylarginine dimethylaminohydrolase activity and nitric oxide production in oxLDL-treated human arterial endothelial cells

Adverse Effects of Cigarette Smoke on NO Bioavailability

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