Insulin‐stimulated <scp>L</scp>‐arginine transport requires <i>SLC7A1</i> gene expression and is associated with human umbilical vein relaxation

González, Marcelo; Gallardo, Victoria; Rodríguez, Natàlia; Salomón, Carlos; Westermeier, Francisco; Guzmán-Gutiérrez, Enrique; Abarzúa, Fernando; Leiva, Andrea; Casanello, Paola; Sobrevía, Luis

doi:10.1002/jcp.22635

Cited by 64 publications

(128 citation statements)

References 35 publications

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“…Furthermore, stimulation of human endothelial cells with insulin has been reported to increase transport of the eNOS substrate L-arginine in an Akt-dependent manner [26]. In agreement with a role for L-arginine transport in insulin-stimulated NO synthesis, inhibition of transport with lysine has been reported to inhibit insulin-stimulated 8 umbilical vein relaxation [30]. Asymmetric dimethylarginine (ADMA) is a naturally occurring, competitive inhibitor of eNOS and elevated ADMA concentrations have been reported in subjects with diabetes [31].…”

Section: Insulin Stimulation Of Endothelial No Synthesismentioning

confidence: 68%

Examining the Role of Insulin in the Regulation of Cardiovascular Health

Salt

2012

Future Cardiol.

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Examining the role of insulin in the regulation of cardiovascular health SUMMARYA substantial body of evidence has reported insulin has direct actions on the cardiovascular system independent of its systemic effects on plasma glucose or lipids. In particular, insulin regulates endothelial synthesis of the vasoactive mediators nitric oxide and endothelin-1, yet the importance of this in the maintenance of cardiovascular health remains poorly understood.Recent studies using animals with targeted downregulation of insulin signaling in vascular tissues are improving our understanding of the role of insulin in vascular health. This article focuses on the direct actions of insulin in cardiovascular tissues, with particular emphasis on the molecular mechanisms of insulin action on endothelial function. The potential contribution of impaired vascular insulin action to the cardiovascular complications of diabetes will also be discussed.

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Section: Insulin Stimulation Of Endothelial No Synthesismentioning

confidence: 68%

Examining the Role of Insulin in the Regulation of Cardiovascular Health

Salt

2012

Future Cardiol.

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“…Because HUVEC from MSPH exhibit unaltered eNOS protein abundance compared with MPH, lower CGRP dilation in MSPH may result from reduced NOS activity. The reduced eNOS activity in MSPH could result from larger reduction of eNOS activationassociated Ser 1177 phosphorylation, 30,38 compared with a less pronounced reduction of eNOS inactivation-associated Thr 495 phosphorylation. 29,39 Furthermore, a larger eNOS inhibition should be expected in MSPH because increased eNOS activity caused by reduced inhibitory Thr 495 phosphorylation could be a phenomenon that will counteract eNOS inactivation as a result of a lower Ser 1177 phosphorylation-dependent stimulation of this enzyme.…”

Section: Vascular Reactivity and Argiimentioning

confidence: 99%

“…ARGII metabolizes l-arginine, the substrate for eNOS in HUVEC 29 and other cell types, 4,21,42 suggesting that a lower l-arginine bioavailability could result in reduced eNOS activity in MSPH. Because l-arginine uptake is required for eNOS activity in HUVEC, 30 a reduced uptake and further delivery of this amino acid to eNOS could occur in MSPH. 43,44 Because HUVEC from MSPH exhibit higher V max and V max /K m for l-arginine transport compared with MPH, a reduced eNOS activity caused by other than reduced l-arginine transport mechanism is likely.…”

Section: Vascular Reactivity and Argiimentioning

confidence: 99%

Maternal Hypercholesterolemia in Pregnancy Associates With Umbilical Vein Endothelial Dysfunction

Leiva

Medina

Salsoso

et al. 2013

ATVB

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Objective-Human pregnancy that courses with maternal supraphysiological hypercholesterolemia (MSPH) correlates with atherosclerotic lesions in fetal arteries. It is known that hypercholesterolemia associates with endothelial dysfunction in adults, a phenomenon where nitric oxide (NO) and arginase are involved. However, nothing is reported on potential alterations in the fetoplacental endothelial function in MSPH. The aim of this study was to determine whether MSPH alters fetal vascular reactivity via endothelial arginase/urea and l-arginine transport/NO signaling pathways. Approach and Results-Total cholesterol <280 mg/dL was considered as maternal physiological hypercholesterolemia (n=46 women) and ≥280 mg/dL as MSPH (n=28 women). Maternal but not fetal total cholesterol and low-density lipoprotein-cholesterol levels were elevated in MSPH. Umbilical veins were used for vascular reactivity assays (wire myography), and primary cultures of umbilical vein endothelial cells to determine arginase, endothelial NO synthase (eNOS), and human cationic amino acid transporter 1 and human cationic amino acid transporter 2A/B expression and activity. MSPH reduced calcitonine gene-related peptide-umbilical vein relaxation and increased intima/media ratio (histochemistry), as well as reduced eNOS activity (l-citrulline synthesis from l-arginine, eNOS phosphorylation/dephosphorylation), but increased arginase activity and arginase II protein abundance. Arginase inhibition increased eNOS activity and l-arginine transport capacity without altering human cationic amino acid transporter 1 or human cationic amino acid transporter 2A/B protein abundance in maternal physiological hypercholesterolemia and MSPH. Conclusions-MSPH is a pathophysiological condition altering umbilical vein reactivity because of fetal endothelial dysfunction associated with arginase and eNOS signaling imbalance. We speculate that elevated maternal circulating cholesterol is a factor leading to fetal endothelial dysfunction, which could have serious consequences to the growing fetus. Maternal Hypercholesterolemia in Pregnancy AssociatesWith Umbilical Vein Endothelial Dysfunction Leiva et al MSPH and Umbilical Vein Endothelial Dysfunction 2445NO synthases. Because NO synthesis depends on l-arginine uptake in human placenta endothelium, 10-17 MSPH could result in altered l-arginine transport and NO synthesis, that is, the l-arginine/NO pathway, in fetal endothelium.l-Arginine uptake occurs predominantly via the human cationic amino acid transporters (hCATs) family.13 hCATs family includes ≥5 members, that is, hCAT-1, hCAT-2A, hCAT-2B, hCAT-3, and hCAT-4, 12,13 of which the high-affinity, lowcapacity hCAT-1 is the main isoform expressed in human umbilical vein endothelial cells (HUVEC), 15,16 a cell type exposed to oxygen-and nutrient-enriched blood (ie, arteriallike blood, reaching fetal circulation).18 Interestingly, altered hCAT-1 activity could result in abnormal NO synthesis in HUVEC. 10,11,15,17 Hypercholesterolemia also associates with reduced endotheli...

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“…The sequences −1606 and −650 bp from the transcription start point of the SLC7A1 gene (GenBank: AL596114) were PCRamplified using Elongase Enzyme System (Invitrogen) and cloned into pGL3-basic reporter system [6,18]. The pGL3-hCAT-1 reporter constructs generated were pGL3-hCAT-1 −1606 and pGL3-hCAT-1…”

Section: Hcat-1 Promoter Cloningmentioning

confidence: 99%

“…Incubation medium was removed and replaced by L-lysinefree Krebs and transport was determined as above [18].…”

Section: L-arginine Transportmentioning

confidence: 99%

Insulin requires A1 adenosine receptors expression to reverse gestational diabetes-increased L-arginine transport in human umbilical vein endothelium

Guzmán-Gutiérrez

Armella

Toledo

et al. 2015

Purinergic Signalling

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Gestational diabetes mellitus (GDM) associates with increased L-arginine transport and extracellular concentration of adenosine in human umbilical vein endothelial cells (HUVECs). In this study we aim to determine whether insulin reverses GDM-increased L-arginine transport requiring adenosine receptors expression in HUVECs. Primary cultured HUVECs from full-term normal (n = 38) and diet-treated GDM (n = 38) pregnancies were used. Insulin effect was assayed on human cationic amino acid transporter 1 (hCAT1) expression (protein, mRNA, SLC7A1 promoter activity) and activity (initial rates of L-arginine transport) in the absence or presence of adenosine receptors agonists or antagonists. A 1 adenosine receptors (A 1 AR) and A 2A AR expression (Western blot, quantitative PCR) was determined. Experiments were done in cells expressing or siRNA-suppressed expression of A 1 AR or A 2A AR. HUVECs from GDM exhibit higher maximal transport capacity (maximal velocity (V max )/ apparent Michaelis Menten constant (K m ), V max /K m ), which is blocked by insulin by reducing the V max to values in cells from normal pregnancies. Insulin also reversed the GDMassociated increase in hCAT-1 protein abundance and mRNA expression, and SLC7A1 promoter activity for the fragment −606 bp from the transcription start point. Insulin effects required A 1 AR, but not A 2A AR expression and activity in this cell type. In the absence of insulin, GDM-increased hCAT-1 expression and activity required A 2A AR expression and activity. HUVECs from GDM pregnancies exhibit a differential requirement of A 1 AR or A 2A AR depending on the level of insulin, a phenomenon that represent a condition where adenosine or analogues of this nucleoside could be acting as helpers of insulin biological effects in GDM.

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Insulin‐stimulated L‐arginine transport requires SLC7A1 gene expression and is associated with human umbilical vein relaxation

Cited by 64 publications

References 35 publications

Examining the Role of Insulin in the Regulation of Cardiovascular Health

Examining the Role of Insulin in the Regulation of Cardiovascular Health

Maternal Hypercholesterolemia in Pregnancy Associates With Umbilical Vein Endothelial Dysfunction

Insulin requires A1 adenosine receptors expression to reverse gestational diabetes-increased L-arginine transport in human umbilical vein endothelium

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