This Article Abstract Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Zeiher, A. M. Articles by Busse, R. Search for Related Content PubMed PubMed Citation Articles by Zeiher, A. M. Articles by Busse, R.

(Circulation Research. 1995;76:980-986.)
© 1995 American Heart Association, Inc.

Articles

Nitric Oxide Modulates the Expression of Monocyte Chemoattractant Protein 1 in Cultured Human Endothelial Cells

Andreas M. Zeiher, Beate Fisslthaler, Beate Schray-Utz, Rudi Busse

From the Center of Physiology (B.F., B.S.-U., R.B.), University of Frankfurt, and the Department of Internal Medicine IV (A.M.Z.), Division of Cardiology, University of Frankfurt (Germany).

Correspondence to Andreas M. Zeiher, MD, Department of Internal Medicine IV, Division of Cardiology, University of Frankfurt, Theodor-Stern-Kai 7, Germany.

	Abstract

Top Abstract Introduction Materials and Methods Results Discussion References

 
Abstract The recruitment of monocytes into the arterial wall is one of the earliest events in the pathogenesis of atherosclerosis. Since monocyte chemoattractant protein 1 (MCP-1) plays a key role in the subendothelial recruitment of monocytes, we tested whether nitric oxide (NO) modulates the expression of MCP-1 in cultured human endothelial cells. Inhibition of basal NO production by N^G-nitro-L-arginine (L-NAG) upregulates endothelial MCP-1 mRNA expression (250±20%) and protein secretion. Exogenous addition of NO dose-dependently decreased MCP-1 mRNA expression and secretion. Changes in MCP-1 mRNA expression and protein secretion were paralleled by corresponding changes in chemotactic activity of cell-conditioned media for monocytes. An MCP-1 antibody reduced monocyte chemotactic activity by 85% and completely abolished the increased monocyte chemotactic activity induced by the inhibition of NO production. Elevation of endothelial cGMP levels had no significant effect on MCP-1 mRNA expression. Inhibition of basal endothelial NO production by L-NAG increased binding activity of a nuclear factor B (NF-B)–like transcriptional regulatory factor, whereas exogenous addition of NO decreased NF-B–like binding activity during stimulation with tumor necrosis factor-. Thus, NO modulates MCP-1 expression and monocyte chemotactic activity secreted by human umbilical vein endothelial cells (HUVECs) in culture. The activation of NF-B–like transcriptional regulatory proteins by inhibition of NO suggests a molecular link between an oxidant-sensitive transcriptional regulatory mechanism and NO synthesis in HUVECs.

Key Words: nitric oxide • nuclear transcription factor • endothelial cells • monocyte chemoattractant protein 1 • atherosclerosis

	Introduction

Top Abstract Introduction Materials and Methods Results Discussion References

 
The focal accumulation of lipid-laden foam cells beneath the endothelium represents the earliest visible manifestation of an atherosclerotic lesion in the arterial wall.^{1 2} Experimental studies in animal models of atherosclerosis have demonstrated that these foam cells are derived from circulating monocytes after their attachment to the arterial endothelium.^{3 4} Thus, the recruitment of monocyte/macrophages into the arterial wall is thought to be one of the earliest events in the pathogenesis of atherosclerosis. Penetration of the vascular endothelium by blood monocytes presumably occurs in response to a gradient of chemotactic factors released from cells of the vascular wall.⁵ One of these chemotactic factors is monocyte chemoattractant protein 1 (MCP-1), a monomeric polypeptide with potent and specific chemoattractant activity for monocytes.^{6 7 8 9} MCP-1 accounts for virtually all of the monocyte chemotactic activity secreted by endothelial cells in vitro.^{10 11}

There is experimental evidence that the endothelium itself becomes dysfunctional during the development of atherosclerotic lesions, a process associated with dramatic alterations in the release of endothelium-derived autacoids.¹² The most important of these autacoids is nitric oxide (NO), which has been shown to continuously modulate vascular tone^{13 14} and to inhibit platelet aggregation and adhesion.¹⁵ Moreover, NO also inhibits smooth muscle cell proliferation,¹⁶ attenuates endothelin generation,¹⁷ and modulates leukocyte adhesion to the endothelium.¹⁸ These effects suggest that NO may play a key role in the prevention of the development of atherosclerotic lesions.

The aim of the present study was to determine whether NO modulates the expression of MCP-1 in cultured human endothelial cells, thereby altering the chemoattractant activity of these cells to monocytes.

	Materials and Methods

Top Abstract Introduction Materials and Methods Results Discussion References

 
Cell Culture
Human umbilical cords were obtained from local hospitals, and human umbilical vein endothelial cells (HUVECs) were isolated as described previously.¹⁹ Cells were grown to confluence on plastic Petri dishes (Greiner) precoated with 25 µg/mL fibronectin in medium 199 (Biochrom) supplemented with 20% heat-inactivated fetal calf serum (Vitromex). N^G-Nitro-L-arginine (L-NAG, Serva), 3-morpholinosydnonimine (SIN-1) and C87-3786 (Cassella AG), tumor necrosis factor- (TNF-, Boehringer Mannheim), atrial natriuretic factor (Sigma Chemical Co), and 8-p-chlorophenylthioguanosine-3',5'-cyclic monophosphate (8-PCPT-cGMP, BioLog) were supplemented without prior change of medium. For long-time incubation, SIN-1 was added every 4 hours.

Detection of cAMP and cGMP Levels
Intracellular cAMP levels in cultured HUVECs were measured as previously described²⁰ by using a commercially available radioimmunoassay kit (Amersham). The amount of cGMP was detected as previously described²¹ by using a radioimmunoassay kit from New England Nuclear. Protein amounts were measured according to Lowry et al.²²

Northern Blot Analysis
Total cellular RNA was extracted from confluently grown HUVECs by acidic guanidinium isothiocyanate.²³ After quantification by measuring absorbance at 260 nm, equal amounts of total RNA were size-fractioned by electrophoresis on a 1.2% formaldehyde agarose gel. RNA was transferred to nylon membranes (Hybond, Amersham-Buchler) and fixed by UV cross-linking (1 minute) and baking at 80°C for 2 hours. Hybridization was performed overnight with a 740-bp Kpn I restriction fragment from the clone pXM-hJE34 (kindly provided by B.J. Rollins) containing the coding region for MCP-1/JE. This DNA fragment was radiolabeled with 30-µCi [-³²P]dCTP (3000 Ci/mmol, Amersham Buchler) by random-prime labeling. The radiolabeled DNA was purified by gel filtration (Nick columns, Pharmacia). The blots were finally washed in 0.1xstandard saline citrate and 0.2% SDS at 55°C.

Autoradiography was performed with Fuji RX film at -70°C with intensifying screens (DuPont de Nemours). For quantification of the RNA amounts loaded, the blots were rehybridized with a radiolabeled cDNA probe specific for 18S rRNA derived from mouse.

Measurement of MCP-1 Secretion
Aliquots of the supernatant from primary HUVEC cultures treated with different stimuli were used for Western blot analysis. Equal amounts of protein were separated by SDS-PAGE²⁴ and transferred to nitrocellulose membranes (Schleicher & Schuell). For detection of MCP-1–specific protein, a polyclonal MCP-1 antiserum (Paesel & Lorei) was used as primary antibody. As secondary antibody, a goat anti-rabbit antibody conjugated to peroxidase (Amersham) was used.

Chemotaxis Assay
Human monocytes were obtained from volunteers by sedimentation of heparinized blood as previously described.²⁵ Monocyte chemotaxis was assessed by a microchamber technique (modified Boyden chamber). In brief, 155 µL endothelial cell–conditioned medium (supplemented with 1% fetal calf serum) was placed in the lower chamber of the modified Boyden chamber, and 100 µL monocyte suspension was placed in the upper compartment. Monocyte suspension was adjusted to 1.2x10⁶ cells per milliliter in PBS. The two compartments were separated by a polycarbonate filter (5-µm pore size, Costar). The chambers were incubated at 37°C for 90 minutes. At the end of incubation, filters were removed, fixed, and stained with Pappenheim stain, and five oil-immersion fields were counted after coding of the samples. For each experiment, N-formyl-methyl-leucyl-phenylalanine (f-MLP, 10 nmol/L, Sigma) was used as a reference chemoattractant.

In some experiments, a neutralizing polyclonal rabbit anti-human MCP-1 antibody (Bibby Dunn Labortechnik) was used.

Gel-Shift Assays
Nuclear extracts from stimulated primary HUVEC cultures were isolated as previously described.²⁶ Briefly, the cells were washed twice with ice-cold HEPES-Tyrode solution and collected by centrifugation. Cells were resuspended in (mmol/L) HEPES 10 (pH 7.9), potassium chloride 10, EDTA 0.1, EGTA 0.1, dithiothreitol 1, and phenylmethylsulfonyl fluoride (PMSF) 0.5, along with 5 µg/mL leupeptin, chymostatin, antipain, pepstatin A, trypsin inhibitor, and aprotinin, and incubated for 15 minutes on ice. After addition of Nonidet P-40 (0.7% final concentration) and vortexing, the lysed cells were centrifuged for 15 seconds at 14 000g. The pellet was resuspended in 20 mmol/L HEPES (pH 7.9), 0.4 mol/L sodium chloride, 1 mmol/L EDTA, 1 mmol/L EGTA, 1 mmol/L dithiothreitol, 1 mmol/L PMSF, and 5 µg/mL of each proteinase inhibitor mix (see above), vigorously shaken for 15 minutes at 4°C, and centrifuged for 5 minutes at 14 000g. The supernatant, containing the nuclear proteins, was used for protein quantification. Double-stranded oligonucleotides containing the sequence of the binding site for nuclear factor B (NF-B) were radiolabeled with 25 µCi [-³²P]ATP (specific activity, 6000 Ci/mmol) by using a 5' end labeling kit from Pharmacia and quantified by Cerenkov counting. Binding conditions were slightly modified according to the method of Sen and Baltimore.²⁷ Nuclear protein (6 µg) was incubated with 5000 counts of labeled oligonucleotide in (mmol/L) HEPES 10 (pH 7.5), sodium chloride 100, EDTA 1, and dithiothreitol 1, along with 5% (vol/vol) glycerol and 2 µg poly(dI/dC) (Pharmacia) for 30 minutes at room temperature. The reaction mixture was loaded on a native 6% polyacrylamide gel buffered with (mmol/L) Tris 89, boric acid 89, and EDTA 2. After vacuum drying, the gel was used for autoradiography at -70°C by using Fuji RX film with intensifying screens (DuPont de Nemours).

	Results

Top Abstract Introduction Materials and Methods Results Discussion References

 
Effects of Cell Culture on Intracellular cGMP Content in HUVECs
To assess the amount of NO produced by HUVECs in culture under basal conditions, intracellular cGMP levels, as an index of NO production, were determined. Fig 1 illustrates that intracellular cGMP levels decreased >10-fold between 16 hours and 40 hours after cell plating. Thus, the analysis of NO-mediated effects on gene expression in cultured HUVECs must take into account the downregulation of endogenous NO production, which is directly related to the time span at which the cells were plated. Consequently, all experiments were carried out within 24 hours after cell plating.

View larger version (13K): [in this window] [in a new window]   Figure 1. Graph showing changes in cGMP content in cultured human umbilical vein endothelial cells after plating as a function of time (mean±SEM, n=8).

Inhibition of Basal NO Production Upregulates MCP-1 Expression
MCP-1 mRNA levels of freshly isolated HUVECs slightly increased 3 hours after culturing and then decreased after 6 hours in culture (Fig 2). Inhibition of NO production by L-NAG increased MCP-1 mRNA levels after a 6-hour incubation with L-NAG (Fig 2). Suppression of endogenous NO production by L-NAG for 24 hours increased MCP-1 mRNA levels to 250±20% (P<.01) of control values (n=6). This increase in MCP-1 mRNA expression was specific for L-NAG, since N^G-nitro-D-arginine had no significant effect on MCP-1 mRNA expression. The L-NAG–induced increase in MCP-1 mRNA was paralleled by an increase in MCP-1 secretion into the supernatant of the HUVEC monolayer, indicating increased production of MCP-1 at the protein level after suppression of endogenous NO synthesis.

View larger version (35K): [in this window] [in a new window]   Figure 2. Northern blot showing time-dependent expression of monocyte chemoattractant protein 1 (MCP-1) mRNA in freshly harvested human umbilical vein endothelial cells with (right side) and without (left side) inhibition of basal nitric oxide synthesis by NG-nitro-L-arginine (L-NAG). 18S mRNA levels (bottom) were used to normalize MCP-1 mRNA levels.

On exposure to L-NAG, HUVECs released considerable amounts of monocyte chemotactic activity, as assessed by induction of monocyte migration across polycarbonate filters. Fig 3 illustrates that monocyte chemotactic activity of HUVEC-conditioned media was significantly increased after 12-hour inhibition of NO synthesis. Monocyte chemotactic activity of conditioned media induced by NO synthesis inhibition was 50% of that induced by exposure of HUVECs to 100 U/mL TNF- (Fig 3).

View larger version (34K): [in this window] [in a new window]   Figure 3. Bar graph showing monocyte chemotactic activity of cell-conditioned media (percentage of control, mean±SEM, n=12). Human umbilical vein endothelial cells were incubated in 1% serum in the absence (control [Ctl]) or presence of NG-nitro-L-arginine (L-NAG, 3x10-5 mol/L), 3-morpholinosydnonimine (SIN-1, 3x10-5, 10-4, and 3x10-4 mol/L), or tumor necrosis factor- (TNFa, 100 U/mL) for 12 hours. The number of monocytes adherent to the polycarbonate filter was counted (five high-power fields [HPFs]), and the value obtained for Ctl conditions was used to normalize chemotactic activity (=100%). L-NAG significantly (P<.01) increased monocyte chemotactic activity, whereas SIN-1 dose-dependently decreased monocyte chemotactic activity (P<.05 at 10-4 mol/L, P<.01 at 3x10-4 mol/L). The effect of L-NAG was 50% of that induced by TNFa. N-Formyl-methyl-leucyl-phenylalanine (f-MLP, 10-8 mol/L) was used to assess unspecific stimulation of monocyte chemotaxis in the modified Boyden chamber.

The addition of a polyclonal MCP-1 antibody to HUVEC-conditioned media dose-dependently reduced the L-NAG–induced increase in chemotactic activity for monocytes, which was completely abolished by 1 µg/mL MCP-1 antibody (Fig 4), indicating that most of the chemotactic activity was due to the release of MCP-1 into the supernatant. The antibody did not affect chemotactic activity of f-MLP (10 nmol/L), excluding nonspecific effects.

View larger version (23K): [in this window] [in a new window]   Figure 4. Bar graph showing monocyte chemotactic activity of cell-conditioned media in the presence (stippled bars) and absence (open bars) of a polyclonal monocyte chemoattractant protein 1 (MCP-1) antibody (AB) (mean±SEM, n=6). AB (1 µg/mL) was added to the cell-conditioned media in the modified Boyden chamber. AB decreased the number of monocytes adherent to the polycarbonate filter (counted for five high-power fields [HPFs]) by 85% compared with control (Ctl) conditions without antibody and completely inhibited the NG-nitro-L-arginine (L-NAG)–induced increase in monocyte chemotaxis. AB had no effect on N-formyl-methyl-leucyl-phenylalanine (f-MLP)–induced monocyte chemotaxis, excluding unspecific effects. TNFa indicates tumor necrosis factor-.

Exogenous NO Decreases MCP-1 Expression
To investigate the effects of exogenously added NO, HUVECs were incubated with the NO donor SIN-1 at varying concentrations and times. Fig 5, left, illustrates that exposure of confluent primary HUVECs to the NO donor SIN-1 for 12 hours dose-dependently decreased MCP-1 mRNA levels. Since SIN-1 has been shown to generate superoxide during its metabolism, we additionally investigated the effects of the NO donor C87-3786, which does not generate superoxide during its metabolism.²⁸ However, identical responses were observed with C87-3786. Incubation with C87-3786 (3x10^-5 mol/L for 8 hours) decreased MCP-1 mRNA levels to 44±15% (n=6, P<.01). The effect of SIN-1 was time dependent, with a maximum decrease in MCP-1 mRNA occurring between 8 and 12 hours of incubation with SIN-1 at 100 µmol/L. The decrease in MCP-1 mRNA expression was paralleled by a decrease in MCP-1 secretion into the supernatant (Fig 5, middle). On exposure of HUVECs to SIN-1, there was also a dose- and time-dependent decrease in monocyte chemotactic activity of HUVEC-conditioned media (Fig 3).

View larger version (29K): [in this window] [in a new window]   Figure 5. Left, Northern blot showing dose-dependent effects of the nitric oxide (NO) donor 3-morpholinosydnonimine (SIN-1) on monocyte chemoattractant protein 1 (MCP-1) mRNA expression in cultured human umbilical vein endothelial cells (HUVECs) after 12-hour exposure. Middle, Western blot showing MCP-1 protein in the supernatant from cultured HUVECs incubated with SIN-1 (10-4 mol/L) or tumor necrosis factor- (TNF, 100 U/mL) for 12 hours. Right, Northern blot showing low-dose (10-5 mol/L) effects of the NO donors SIN-1 and C87-3786 on MCP-1 mRNA expression in cultured HUVECs with inhibition of basal NO synthesis by NG-nitro-L-arginine (L-NAG) after 5-hour (short-time) exposure.

Incubation with SIN-1 or C87-3786 at lower concentrations (10^-5 mol/L) for shorter periods (5 hours) did not affect basal MCP-1 mRNA levels. However, it significantly (P<.05) reduced the L-NAG–induced increase in MCP-1 mRNA levels to 51±9% for SIN-1 (n=4) and to 43±12% for C87-3786 (n=4) (Fig 5, right).

Effects of Cyclic Nucleotides on MCP-1 mRNA Levels
The soluble guanylyl cyclase is the major molecular target of NO in endothelial cells. Therefore, we investigated whether NO-induced modulation of MCP-1 expression is mediated by changes in cGMP levels. Fig 6 illustrates that increased levels of cGMP caused either by treatment of HUVECs with atrial natriuretic factor (3.4±1.3-fold increase in cGMP levels) or by the addition of the stable membrane-permeable cGMP analogue 8-PCPT-cGMP had no effect on MCP-1 mRNA levels. Thus, elevated cGMP levels may not account for the decrease in MCP-1 mRNA levels after exogenous NO supplementation.

View larger version (51K): [in this window] [in a new window]   Figure 6. Northern blot demonstrating monocyte chemoattractant protein 1 (MCP-1) mRNA levels after elevation of cGMP levels in human umbilical vein endothelial cells by incubation with 8-p-chlorophenylthioguanosine-3',5'-cyclic monophosphate (8-PCPT-cGMP, 100 µmol/L) or atrial natriuretic factor (ANF, 30 nmol/L) for 12 hours.

In contrast, elevation of cAMP levels by treatment of HUVECs with isoproterenol (3.8±1-fold increase in cAMP levels) and the phosphodiesterase inhibitor isobutyl methylxanthine induced an increase in MCP-1 mRNA levels (Fig 7), indicating that elevations of intracellular cAMP levels constitute a potential signaling mechanism involved in the induction of MCP-1 in endothelial cells. However, inhibition of endogenous NO synthesis by L-NAG decreased intracellular cAMP levels to 50% to 60% of control values (data not shown), suggesting that the increase in MCP-1 mRNA levels by inhibition of endogenous NO synthesis cannot be explained by increased intracellular levels of cAMP.

View larger version (105K): [in this window] [in a new window]   Figure 7. Northern blot demonstrating monocyte chemoattractant protein 1 (MCP-1) mRNA levels after elevation of cAMP levels in human umbilical vein endothelial cells by incubation with isoproterenol (Iso, 100 µmol/L), isobutyl methylxanthine (IBMX, 500 µmol/L), or Iso (100 µmol/L) plus IBMX (500 µmol/L) for 12 hours. MCP-1 mRNA levels increased by 198±38% after incubation with Iso (n=4), by 201±29% after combined incubation with Iso and IBMX (n=4), and by 137±15% after incubation with IBMX (n=3).

NO Inhibition Promotes DNA Binding Protein Activity to NF-B–Like Binding Sites
Since the promoter of the human MCP-1 gene contains functionally active NF-B binding sites,²⁹ we tested whether an NF-B–like transcription factor is involved in the effects of NO inhibition. Nuclear extracts from HUVECs were assayed for DNA binding activity to a double-stranded oligonucleotide containing the sequence for an NF-B binding site. As shown in Fig 8, NF-B–like binding activity was increased in response to inhibition of NO production by L-NAG. TNF- further induced NF-B binding activity. However, costimulation with the NO donor C87-3786 produced a decrease in NF-B–like binding activity compared with stimulation with TNF- alone (Fig 8). Quantitative densitometric analysis of the gel-shift assays revealed that L-NAG increased NF-B binding activity by 150±10.7% (n=5), whereas TNF- led to an increase of 345±44% (n=4). Costimulation with the NO donor SIN-1 decreased TNF-–induced binding activity by 68±7.8% (n=3); costimulation with the NO donor C87-3786, by 75±13.3% (n=4). The specificity for NF-B was shown by competition of the gel shift with unlabeled NF-B oligonucleotide and the inhibition of the shift by the inhibitory NF-B subunit IB (data not shown). Thus, inhibition of NO production upregulates the binding activity of an NF-B–like transcription factor in HUVECs.

View larger version (59K): [in this window] [in a new window]   Figure 8. Gel mobility shift assay demonstrating effect of nitric oxide on the activation of nuclear factor B (NF-B). Human umbilical vein endothelial cells were treated for 30 minutes with the components indicated: NG-nitro-L-arginine (L-NAG, 3x10-5 mol/L), tumor necrosis factor- (TNF-, 1000 U/mL), and C87-3786 (10-4 mol/L). Equal amounts of proteins from nuclear extract were analyzed for NF-B binding activity. The signal indicated is specific for NF-B, since inhibition with the inhibitory NF-B subunit IB abolished the signal (data not shown).

	Discussion

Top Abstract Introduction Materials and Methods Results Discussion References

 
The mechanisms responsible for the recruitment of monocytes into the arterial wall during atherogenesis are believed to involve the adherence of monocytes to the endothelium, followed by their migration into the subendothelial space. Previous studies of transmigration have demonstrated that virtually all of the monocyte chemoattractant activity released into the culture medium by human endothelial cells was attributable to MCP-1.¹⁰ In addition, MCP-1 antibodies completely inhibited the minimally modified low-density lipoprotein (LDL)–induced monocyte transmigration into the subendothelial space of a coculture system of human aortic wall cells.¹⁰ Thus, MCP-1 appears to play a key role in the recruitment of monocytes. The present study demonstrates that both endogenous and exogenous NO modulate the expression of MCP-1 in human endothelial cells in vitro. This was exemplified by the finding that inhibition of endogenous NO production markedly increased MCP-1 mRNA levels in endothelial cells. Moreover, the chemotactic response of monocytes to medium from cultured endothelial cells was significantly enhanced after inhibition of endothelial NO synthesis. The latter effect can be attributed solely to an increase in MCP-1, since MCP-1 antibodies completely prevented this enhanced monocyte chemotactic activity. In addition, exogenous NO induced a decrease in both MCP-1 expression and monocyte chemoattractant activity of cell-conditioned media.

Previous studies of MCP-1 expression by endothelial cells focused primarily on the effects of cytokines^{11 30} or modified LDL.¹⁰ Since atherosclerotic lesions have been shown to contain oxidatively modified LDL³¹ and inflammatory cells capable of secreting a number of cytokines,² induction of MCP-1 expression by cytokines and modified LDL within the vascular wall was regarded to be important for the monocyte recruitment into the subendothelium. Indeed, in situ hybridization studies have demonstrated increased MCP-1 mRNA expression in atherosclerotic lesions from both humans and experimental animals.^{32 33 34} In these studies, MCP-1 was detected not only in endothelial cells and macrophages but also in smooth muscle cells of atherosclerotic vessels.^{33 34} These results suggested that the recruitment of peripheral blood monocytes into the subintimal space occurs, in part, in response to a chemotactic gradient of MCP-1 released from early foam cell lesions and smooth muscle cells.³⁴ The present study considerably extends these findings by demonstrating that NO modulates MCP-1 expression and monocyte chemotactic activity in vitro irrespective of the presence or absence of cytokines or modified LDL.

It is now well documented that NO production in endothelial cells in situ is closely correlated to the shear stress prevailing on the endothelial cell surface.^{35 36 37} Low shear stress is associated with an attenuated NO synthesis; increases in shear stress induce not only an immediate increase in NO synthesis but also chronically enhance NO production by increasing NO synthase expression.³⁶ Therefore, it is tempting to speculate that since arterial sites with low shear stress or flow turbulences are the locations in which the earliest grossly visible atherosclerotic lesions typically develop,³⁸ these sites may exhibit an enhanced expression of MCP-1. Our results demonstrate that inhibition of endogenous NO formation potently induces MCP-1 in human endothelial cells in vitro.

It has been shown that shear stress is associated with a transient initial increase in MCP-1 gene expression.³⁹ However, this initial increase in MCP-1 mRNA expression is unrelated to the magnitude of shear stress and not confined to endothelial cells but can also be observed in HeLa cells and glioma cells.³⁹ Thus, this initial increase in MCP-1 mRNA most likely represents an unspecific stress response with subsequent induction of immediate-early response genes such as MCP-1 mediated by a protein kinase C–dependent mechanism. This view is also supported by our observation of an initial increase in MCP-1 mRNA shortly after culturing freshly isolated HUVECs (Fig 2). However, in addition to this early increase in MCP-1 mRNA in untreated HUVECs, inhibition of NO synthesis induced a pronounced enhancement of MCP-1 mRNA expression, which was still present 24 hours after culturing the cells. Thus, the mechanisms responsible for MCP-1 gene expression in response to inhibition of NO synthesis must be different from those inducing an immediate-early response to any kind of stress.

Circumstantial evidence suggests that NO might exert important antiatherosclerotic effects. For example, NO has been reported to inhibit vascular smooth muscle cell proliferation,¹⁶ and removal of the endothelium has been shown to induce considerable intimal thickening of the vessel wall.⁴⁰ High-flow states, which are associated with a concomitant increased endothelial NO production on the other hand, reduce intimal thickening even after arterial injury in experimental animals.⁴¹ Finally, supplementation of L-arginine, the precursor of NO, has been recently shown not only to attenuate endothelial adhesiveness for monocytes⁴² but also to reduce the extent of atherosclerotic lesions in cholesterol-fed rabbits.⁴³ The modulation of MCP-1 in endothelial cells by endogenously produced NO might importantly contribute to the antiatherogenic effects of NO by locally interfering with the monocyte chemotactic activity of the endothelial cell layer itself.

The molecular mechanisms underlying the modulation of MCP-1 expression in endothelial cells by NO remain to be determined. Previous studies addressing potential signaling mechanisms involved in MCP-1 induction in fibroblasts and vascular smooth muscle cells demonstrated a role for protein kinase C, whereas cAMP did not appear to be involved.^{44 45} The results of the present study and a recent report⁴⁶ clearly demonstrate that increased levels of intracellular cAMP induce MCP-1 expression in human endothelial cells. Thus, the signaling mechanisms involved in MCP-1 expression may be different in different cell types. However, a cAMP-mediated mechanism cannot account for the observed increase in MCP-1 expression after inhibition of NO synthesis, since incubation of endothelial cells with L-NAG led to a decrease in cAMP levels.

Activation of soluble guanylyl cyclase, the major target of NO in a variety of cells, results in an increase in cGMP. However, increased levels of intracellular cGMP did not affect MCP-1 expression in HUVECs, indicating that the modulation of MCP-1 by NO is obviously not mediated by intracellular cGMP levels. In line with our results, Niu et al⁴⁷ recently reported that prolonged exposure of endothelial cells to inhibitors of NO synthesis induces neutrophil adhesion by a mechanism independent of cGMP.

The molecular targets of NO in endothelial cells include not only the activation of soluble guanylyl cyclase but also the activation of an ADP-ribosyltransferase, the formation of nitrosylated Fe-S complexes with potential destruction of the catalytic function of some enzymes, and interactions with superoxide anions.⁴⁸ Depending on circumstances, the reaction of superoxide with NO may represent a detoxification of either molecule or a route to the generation of peroxynitrite, which might decay to hydroxyl radical,⁴⁹ considered the strongest oxidant in biological systems. Thus, NO may either scavenge oxygen radicals or, if present in large quantities, act as a radical by itself.⁴⁸ Indeed, inhibition of NO formation in HUVECs has recently been shown to increase oxidative flux within these cells,⁴⁷ indicating that endogenous NO may function to scavenge endogenously produced oxidants. Reactive oxygen intermediates have been recognized to play an important role as widely used regulators of gene transcription,^{50 51} especially for genes under the control of the transcription factor NF-B.⁵⁰ The promoter of the human MCP-1 gene contains functionally active NF-B binding sites.²⁹ In mesangial cells, oxygen radicals may serve as second messengers for the expression of MCP-1 in response to TNF- and IgG.⁵² Moreover, cytokine-induced oxidative stress is an important regulatory signal in endothelial cells that mediates the expression of vascular cell adhesion molecule-1 via NF-B–like transcriptional regulatory proteins.⁵³ The present study demonstrates that inhibition of NO synthesis activates proteins capable of binding to oligonucleotides containing the consensus sequence for an NF-B binding site, suggesting a molecular link between an oxidant-sensitive transcriptional regulatory mechanism and NO synthesis in HUVECs. However, we cannot exclude the possibility that other transcription factors such as activator protein 1, which has recently been characterized as an antioxidant-responsive factor,⁵⁴ may play a role as regulatory signals modulating MCP-1 expression during inhibition of NO synthesis in HUVECs. Thus, although there is accumulating evidence that NO plays an important role in the maintenance of cellular redox equilibrium, the precise mechanism(s) by which NO interferes with other oxidants and modulates the expression of MCP-1 in HUVECs will require further studies.

In summary, endogenously produced NO modulates MCP-1 expression and secretion in primary cultures of HUVECs. Since endothelial NO production is closely associated with the shear stress acting on the endothelial cell layer, upregulation of MCP-1 expression by reduced NO production might play a key role in the preferential development of early atherosclerotic lesions at arterial sites with low shear stress by locally increasing the monocyte chemotactic activity of the endothelium.

	Acknowledgments

 
This study was supported by the Deutsche Forschungsgemeinschaft (grant Bu 436/4-3). The authors are indebted to Isabel Winter and Christine Kremer for expert technical assistance.

Received December 5, 1994; accepted March 15, 1995.

	References

Top Abstract Introduction Materials and Methods Results Discussion References

 
1. Ross R. The pathogenesis of atherosclerosis. N Engl J Med. 1986;314:488-500.[Medline] [Order article via Infotrieve]

2. Munro JM, Cotran RS. The pathogenesis of atherosclerosis: atherogenesis and inflammation. Lab Invest. 1988;58:249-261. [Medline] [Order article via Infotrieve]

3. Faggiotto A, Ross R, Harker L. Studies of hypercholesterolemia in the nonhuman primate, I: changes that lead to fatty streak formation. Arteriosclerosis. 1984;4:323-340. [Abstract/Free Full Text]

4. Gerrity R. The role of the monocyte in atherogenesis, I: transition of blood-borne monocytes into foam cells in fatty lesions. Am J Pathol. 1983;103:181-190.[Abstract]

5. Navab M, Imes SS, Hama SY, Hough GP, Ross LA, Bork RW, Valente AJ, Berliner JA, Drinkwater DC, Laks H, Fogelman AM. Monocyte transmigration induced by modification of low density lipoprotein in cocultures of human aortic wall cells is due to induction of monocyte chemotactic protein 1 synthesis and is abolished by high density lipoprotein. J Clin Invest. 1991;88:2039-2046.

6. Yoshimura T, Robinson EA, Tanaka S, Appella E, Kuratsu J, Leonard EJ. Purification and amino acid analysis of two human glioma-derived monocyte chemoattractants. J Exp Med. 1989;169:1449-1459. [Abstract/Free Full Text]

7. Yoshimura T, Yukhi N, Moore SK, Appella E, Lerman MI, Leonard EJ. Human monocyte chemoattractant protein-1 (MCP-1). FEBS Lett. 1989;244:487-493. [Medline] [Order article via Infotrieve]

8. Rollins BJ, Stier P, Ernst T, Wong GG. Human JE encodes a monocyte secretory protein. Mol Cell Biol. 1989;9:4687-4695. [Abstract/Free Full Text]

9. Graves DT, Jliang YL, Williamson MJ, Valente AJ. Identification of monocyte chemotactic activity produced by malignant cells. Science. 1989;245:1490-1493. [Abstract/Free Full Text]

10. Cushing SD, Berliner JA, Valente AJ, Territo MC, Navab M, Parhami F, Gerrity R, Schwartz CJ, Fogelman AM. Minimally modified low density lipoprotein induces monocyte chemotactic protein 1 in human endothelial cells and smooth muscle cells. Proc Natl Acad Sci U S A. 1990;87:5134-5138. [Abstract/Free Full Text]

11. Rollins BJ, Yoshimura T, Leonard EJ, Pober JS. Cytokine-activated human endothelial cells synthesize and secrete a monocyte chemoattractant, MCP-1/JE. Am J Pathol. 1990;136:1229-1233. [Abstract]

12. Freiman PC, Mitchell GC, Heistad DD, Armstrong ML, Harrison DG. Atherosclerosis impairs endothelium-dependent vascular relaxation to acetylcholine and thrombin in primates. Circ Res. 1986;58:783-789. [Abstract/Free Full Text]

13. Ignarro JJ. Biosynthesis and metabolism of endothelium-derived nitric oxide. Annu Rev Pharmacol Toxicol. 1990;30:535-560. [Medline] [Order article via Infotrieve]

14. Bassenge E, Busse R. Endothelial modulation of coronary tone. Prog Cardiovasc Dis. 1988;30:349-380. [Medline] [Order article via Infotrieve]

15. Busse R, Lückhoff A, Bassenge E. Endothelium-derived relaxant factor inhibits platelet activation. Naunyn Schmiedebergs Arch Pharmacol. 1987;336:566-571. [Medline] [Order article via Infotrieve]

16. Garg UC, Hassid A. Nitric-oxide generating vasodilators and 8-bromo-cyclic guanosine monophosphate inhibit mitogenesis and proliferation of cultured rat vascular smooth muscle cells. J Clin Invest. 1989;83:1774-1777.

17. Boulanger C, Lüscher T. Release of endothelin from the porcine aorta. J Clin Invest. 1990;85:587-590.

18. Kubes P, Suzuki M, Granger DN. Nitric oxide: an endogenous modulator of leukocyte adhesion. Proc Natl Acad Sci U S A. 1991;88:4651-4655. [Abstract/Free Full Text]

19. Busse R, Lamontagne D. Endothelium-derived bradykinin is responsible for the increase in calcium produced by angiotensin-converting enzyme inhibitors in human endothelial cells. Naunyn Schmiedebergs Arch Pharmacol. 1991;344:126-129. [Medline] [Order article via Infotrieve]

20. Lückhoff A, Mülsch A, Busse R. cAMP attenuates autacoid release from endothelial cells: relation to internal calcium. Am J Physiol. 1990;258:H960-H966. [Abstract/Free Full Text]

21. Mülsch A, Böhme E, Busse R. Stimulation of soluble guanylate cyclase by endothelium-derived relaxing factor from cultured endothelial cells. Eur J Pharmacol. 1987;135:247-250. [Medline] [Order article via Infotrieve]

22. Lowry OH, Rosebrough MJ, Farr AL, Randall RJ. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951;193:265-275. [Free Full Text]

23. Chomcynzki P, Sacchi N. Single step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987;162:156-159. [Medline] [Order article via Infotrieve]

24. Laemmli UK. Cleavage of structural proteins during the assembly of head of bacteriophage T4. Nature. 1970;227:680-685. [Medline] [Order article via Infotrieve]

25. Wang JM, Sica A, Peri G, Walter S, Padura M, Libby P, Ceska M, Lindley I, Colotta F, Mantovani A. Expression of monocyte chemotactic protein and interleukin-8 by cytokine-activated human vascular smooth muscle cells. Arterioscler Thromb. 1991;11:1166-1174. [Abstract/Free Full Text]

26. Schreiber E, Matthias P, Müller MM, Schaffner W. Rapid detection of octamer binding proteins with `mini-extracts,' prepared from a smaller number of cells. Nucleic Acids Res. 1989;17:6419. [Free Full Text]

27. Sen R, Baltimore D. Multiple nuclear factors interact with the immunoglobulin enhancer sequence. Cell. 1986;46:705-715. [Medline] [Order article via Infotrieve]

28. Mülsch A, Hecker M, Mordvintcev PI, Vanin AF, Busse R. Enzymic and nonenzymic release of NO accounts for the vasodilator activity of the metabolites of CAS 936, a novel long-acting sydnonimine derivative. Naunyn Schmiedebergs Arch Pharmacol. 1993;347:92-100. [Medline] [Order article via Infotrieve]

29. Uead A, Okuda K, Ohno S, Shirai A, Igarashi T, Matsunaga K, Fukushima J, Kawamoto S, Ishigatsubo J, Okubo T. NF-B and Sp1 regulate transcription of the human monocyte chemoattractant protein-1 gene. J Immunol. 1994;153:2052-2063. [Abstract]

30. Strieter RM, Wiggins R, Phan SH, Wharram BL, Showell HJ, Remick DG, Chensue SW, Kunkel SL. Monocyte chemotactic protein gene expression by cytokine-treated human fibroblasts and endothelial cells. Biochem Biophys Res Commun. 1989;162:694-700. [Medline] [Order article via Infotrieve]

31. Ylä-Herttuala S, Palinski W, Rosenfeld ME, Parthasarathy C, Carew TE, Butler S, Witzum JL, Steinberg D. Evidence for the presence of oxidatively modified low density lipoprotein in atherosclerotic lesions of rabbit and man. J Clin Invest. 1989;84:1086-1095.

32. Ylä-Herttuala S, Lipton BA, Rosenfeld ME, Särkioja T, Yoshimura T, Leonard EJ, Witztum JL, Steinberg D. Expression of monocyte chemoattractant protein 1 in macrophage-rich areas of human and rabbit atherosclerotic lesions. Proc Natl Acad Sci U S A. 1991;88:5252-5256. [Abstract/Free Full Text]

33. Nelken NA, Coughlin SR, Gordon D, Wilcox JN. Monocyte chemoattractant protein-1 in human atheromatous plaques. J Clin Invest. 1991;88:1121-1127.

34. Yu X, Dluz S, Graves DT, Zhang L, Antoniades HA, Hollander W, Prusty S, Valente AJ, Schwartz CJ, Sonenshein GE. Elevated expression of monocyte chemoattractant protein 1 by vascular smooth muscle cells in hypercholesterolemic primates. Proc Natl Acad Sci U S A. 1992;89:6953-6957. [Abstract/Free Full Text]

35. Busse R, Pohl U. Fluid shear-stress-dependent stimulation of endothelial autacoid release: mechanisms and significance for the control of vascular tone. In: Frangos JA, ed. Physical Forces and the Mammalian Cell. San Diego, Calif: Academic Press Inc; 1993:223-248.

36. Sessa WC, Pritchard K, Seyedi N, Wang J, Hintze TH. Chronic exercise in dogs increases coronary vascular nitric oxide production and endothelial cell nitric oxide synthase gene expression. Circ Res. 1994;74:349-353.[Abstract/Free Full Text]

37. Nishida K, Harrison DG, Navas JP, Fisher AA, Dockery SP, Uematsu M, Nerem RM, Alexander RW, Murphy TJ. Molecular cloning and characterization of the constitutive bovine aortic endothelial nitric oxide synthase. J Clin Invest. 1992;90:2092-2096.

38. Ku DN, Giddens DP, Zarins CK, Glagov S. Pulsatile flow and atherosclerosis in the human carotid bifurcation: positive correlation between plaque location and low oscillating shear stress. Arteriosclerosis. 1985;5:293-302. [Abstract/Free Full Text]

39. Shyy Y-J, Hsieh H-J, Usami S, Chien S. Fluid shear stress induces a biphasic response of human monocyte chemotactic protein 1 gene expression in vascular endothelium. Proc Natl Acad Sci U S A. 1994;91:4678-4682. [Abstract/Free Full Text]

40. Langille BL, O'Donnell T. Reduction in arterial diameter produced by chronic decreases in blood flow are endothelium-dependent. Science. 1986;231:405-407. [Abstract/Free Full Text]

41. Reidy MA, Fingerle J, Lindner V. Factors controlling the development of arterial lesions after injury. Circulation. 1992;86(suppl III):III-43-III-46.

42. Tsao PS, McEvoy LM, Drexler H, Butcher EC, Cooke JP. Enhanced endothelial adhesiveness in hypercholesterolemia is attenuated by L-arginine. Circulation. 1994;89:2176-2182. [Abstract/Free Full Text]

43. Cooke JP, Singer AH, Tsao P, Zear P, Rowan RA, Billingham ME. Antiatherogenic effects of L-arginine in the hypercholesterolemic rabbit. J Clin Invest. 1992;90:1168-1172.

44. Taubmann MB, Rollins BJ, Poon M, Marmur J, Green RS, Berk BC, Nadal-Ginard B. JE mRNA accumulates rapidly in aortic injury and in platelet-derived growth factor–stimulated vascular smooth muscle cells. Circ Res. 1992;70:314-325. [Abstract/Free Full Text]

45. Hall DJ, Stiles CD. Platelet-derived growth factor-inducible genes respond differentially to at least two distinct intracellular messengers. J Biol Chem. 1987;262:15302-15308. [Abstract/Free Full Text]

46. Shyy Y, Li Y, Kolattukudy PE. Activation of MCP-1 gene expression is mediated through multiple signalling pathways. Biochem Biophys Res Commun. 1993;192:693-697. [Medline] [Order article via Infotrieve]

47. Niu X, Smith CW, Kubes P. Intracellular oxidative stress induced by nitric oxide synthesis inhibition increases endothelial cell adhesion to neutrophils. Circ Res. 1994;74:1133-1140. [Abstract/Free Full Text]

48. Nathan C. Nitric oxide as a secretory product of mammalian cells. FASEB J. 1992;6:3051-3064. [Abstract]

49. Beckman JS, Beckman TW, Chen J, Marshall PA, Freeman BA. Apparent hydroxyl radical production by peroxynitrite: implications for endothelial injury from nitric oxide and superoxide. Proc Natl Acad Sci U S A. 1990;87:1620-1624. [Abstract/Free Full Text]

50. Schreck R, Rieber P, Bäuerle PA. Reactive oxygen intermediates as apparently widely used second messengers in the activation of the NF-B transcription factor and HIV-1. EMBO J. 1991;10:2247-2258. [Medline] [Order article via Infotrieve]

51. Toledamo MB, Leonhard WJ. Modulation of transcription factor NF-B binding by oxidation-reduction in vitro. Proc Natl Acad Sci U S A. 1991;88:4328-4332. [Abstract/Free Full Text]

52. Satriano JA, Shuldiner M, Hora K, Zing Y, Shan Z, Schlondorff D. Oxygen radicals as second messengers for expression of the monocyte chemoattractant protein, JE/MCP-1, and the monocyte colony-stimulating factor, CSF-1, in response to tumor necrosis factor- and immunoglobulin G. J Clin Invest. 1993;92:1546-1571.

53. Marui N, Offermann MK, Swerlick R, Kunsch C, Rosen CA, Ahmad M, Alexander RW, Medford RM. Vascular cell adhesion molecule-1 (VCAM-1) gene transcription and expression are regulated through an antioxidant-sensitive mechanism in human vascular endothelial cells. J Clin Invest. 1993;92:1866-1874.

54. Meyer M, Schreck R, Baeuerle PA. H₂O₂ and antioxidants have opposite effects on activation of NF-B and AP-1 in intact cells: AP-1 as secondary antioxidant-responsive factor. EMBO J. 1993;12:2005-2015.[Medline] [Order article via Infotrieve]

This article has been cited by other articles:

				H.-C. Ou, W.-J. Lee, I-T. Lee, T.-H. Chiu, K.-L. Tsai, C.-Y. Lin, and W. H.-H. Sheu Ginkgo biloba extract attenuates oxLDL-induced oxidative functional damages in endothelial cells J Appl Physiol, May 1, 2009; 106(5): 1674 - 1685. [Abstract] [Full Text] [PDF]

				E. Demicheva, M. Hecker, and T. Korff Stretch-Induced Activation of the Transcription Factor Activator Protein-1 Controls Monocyte Chemoattractant Protein-1 Expression During Arteriogenesis Circ. Res., August 29, 2008; 103(5): 477 - 484. [Abstract] [Full Text] [PDF]

				L. G. Spagnoli, E. Bonanno, G. Sangiorgi, and A. Mauriello Role of Inflammation in Atherosclerosis J. Nucl. Med., November 1, 2007; 48(11): 1800 - 1815. [Abstract] [Full Text] [PDF]

				G. Dever, C. M. Spickett, S. Kennedy, C. Rush, G. Tennant, A. Monopoli, and C. L. Wainwright The Nitric Oxide-Donating Pravastatin Derivative, NCX 6550 [(1S-[1{alpha}(betaS*, {delta}S*), 2{alpha}, 6{alpha}, 8beta-(R*), 8a{alpha}]]-1,2,6,7,8,8a-Hexahydro-beta, {delta}, 6-trihydroxy-2-methyl-8-(2-methyl-1-oxobutoxy)-1-naphtalene-heptanoic Acid 4-(Nitrooxy)butyl Ester)], Reduces Splenocyte Adhesion and Reactive Oxygen Species Generation in Normal and Atherosclerotic Mice J. Pharmacol. Exp. Ther., January 1, 2007; 320(1): 419 - 426. [Abstract] [Full Text] [PDF]

				T. Hayashi, T. Esaki, D. Sumi, T. Mukherjee, A. Iguchi, and G. Chaudhuri Modulating role of estradiol on arginase II expression in hyperlipidemic rabbits as an atheroprotective mechanism PNAS, July 5, 2006; 103(27): 10485 - 10490. [Abstract] [Full Text] [PDF]

				A. G. Huebschmann, J. G. Regensteiner, H. Vlassara, and J. E.B. Reusch Diabetes and Advanced Glycoxidation End Products. Diabetes Care, June 1, 2006; 29(6): 1420 - 1432. [Full Text] [PDF]

				B. Lorkowska, M. Bartus, M. Franczyk, R. B. Kostogrys, J. Jawien, P. M. Pisulewski, and S. Chlopicki Hypercholesterolemia Does Not Alter Endothelial Function in Spontaneously Hypertensive Rats J. Pharmacol. Exp. Ther., June 1, 2006; 317(3): 1019 - 1026. [Abstract] [Full Text] [PDF]

				K. Takenaka, Y. Nishimura, T. Nishiuma, A. Sakashita, T. Yamashita, K. Kobayashi, M. Satouchi, T. Ishida, S. Kawashima, and M. Yokoyama Ventilator-induced lung injury is reduced in transgenic mice that overexpress endothelial nitric oxide synthase Am J Physiol Lung Cell Mol Physiol, June 1, 2006; 290(6): L1078 - L1086. [Abstract] [Full Text] [PDF]

				A. G. Herman and S. Moncada Therapeutic potential of nitric oxide donors in the prevention and treatment of atherosclerosis Eur. Heart J., October 1, 2005; 26(19): 1945 - 1955. [Abstract] [Full Text] [PDF]

				J. P. Cullen, S. Sayeed, Y. Jin, N. G. Theodorakis, J. V. Sitzmann, P. A. Cahill, and E. M. Redmond Ethanol inhibits monocyte chemotactic protein-1 expression in interleukin-1{beta}-activated human endothelial cells Am J Physiol Heart Circ Physiol, October 1, 2005; 289(4): H1669 - H1675. [Abstract] [Full Text] [PDF]

				T. Sakamoto, T. Ishibashi, N. Sakamoto, K. Sugimoto, K. Egashira, H. Ohkawara, K. Nagata, K. Yokoyama, M. Kamioka, T. Ichiki, et al. Endogenous NO Blockade Enhances Tissue Factor Expression via Increased Ca2+ Influx Through MCP-1 in Endothelial Cells by Monocyte Adhesion Arterioscler Thromb Vasc Biol, September 1, 2005; 25(9): 2005 - 2011. [Abstract] [Full Text] [PDF]

				K.-i. Kodama, Y. Nishio, O. Sekine, Y. Sato, K. Egawa, H. Maegawa, and A. Kashiwagi Bidirectional regulation of monocyte chemoattractant protein-1 gene at distinct sites of its promoter by nitric oxide in vascular smooth muscle cells Am J Physiol Cell Physiol, September 1, 2005; 289(3): C582 - C590. [Abstract] [Full Text] [PDF]

				V. Mollace, C. Muscoli, E. Masini, S. Cuzzocrea, and D. Salvemini Modulation of Prostaglandin Biosynthesis by Nitric Oxide and Nitric Oxide Donors Pharmacol. Rev., June 1, 2005; 57(2): 217 - 252. [Abstract] [Full Text] [PDF]

				H. Homma, E. Hoy, D.-Z. Xu, Q. Lu, R. Feinman, and E. A. Deitch The female intestine is more resistant than the male intestine to gut injury and inflammation when subjected to conditions associated with shock states Am J Physiol Gastrointest Liver Physiol, March 1, 2005; 288(3): G466 - G472. [Abstract] [Full Text] [PDF]

				D. H. Endemann and E. L. Schiffrin Endothelial Dysfunction J. Am. Soc. Nephrol., August 1, 2004; 15(8): 1983 - 1992. [Abstract] [Full Text] [PDF]

				S. M. Boekholdt, R. J. G. Peters, C. E. Hack, N. E. Day, R. Luben, S. A. Bingham, N. J. Wareham, P. H. Reitsma, and K.-T. Khaw IL-8 Plasma Concentrations and the Risk of Future Coronary Artery Disease in Apparently Healthy Men and Women: The EPIC-Norfolk Prospective Population Study Arterioscler Thromb Vasc Biol, August 1, 2004; 24(8): 1503 - 1508. [Abstract] [Full Text] [PDF]

				K. K. Koh, J. W. Son, J. Y. Ahn, D. S. Kim, D. K. Jin, H. S. Kim, S. H. Han, Y.-H. Seo, W.-J. Chung, W. C. Kang, et al. Simvastatin Combined With Ramipril Treatment in Hypercholesterolemic Patients Hypertension, August 1, 2004; 44(2): 180 - 185. [Abstract] [Full Text] [PDF]

				U. Landmesser, B. Hornig, and H. Drexler Endothelial Function: A Critical Determinant in Atherosclerosis? Circulation, June 1, 2004; 109(21_suppl_1): II-27 - II-33. [Abstract] [Full Text] [PDF]

				N. C. Weber, S. B. Blumenthal, T. Hartung, A. M. Vollmar, and A. K. Kiemer ANP inhibits TNF-{alpha}-induced endothelial MCP-1 expression--involvement of p38 MAPK and MKP-1 J. Leukoc. Biol., November 1, 2003; 74(5): 932 - 941. [Abstract] [Full Text] [PDF]

				M. A. Creager, T. F. Luscher, F. Cosentino, and J. A. Beckman Diabetes and Vascular Disease: Pathophysiology, Clinical Consequences, and Medical Therapy: Part I Circulation, September 23, 2003; 108(12): 1527 - 1532. [Full Text] [PDF]

				K. K. Koh, J. Y. Ahn, S. H. Han, D. S. Kim, D. K. Jin, H. S. Kim, M.-S. Shin, T. H. Ahn, I. S. Choi, and E. K. Shin Pleiotropic effects of angiotensin II receptor blocker in hypertensive patients J. Am. Coll. Cardiol., September 3, 2003; 42(5): 905 - 910. [Abstract] [Full Text] [PDF]

				W. Schaper and D. Scholz Factors Regulating Arteriogenesis Arterioscler Thromb Vasc Biol, July 1, 2003; 23(7): 1143 - 1151. [Abstract] [Full Text] [PDF]

				J. M. Proudfoot, K. D. Croft, I. B. Puddey, and L. J. Beilin Angiotensin II Type 1 Receptor Antagonists Inhibit Basal As Well As Low-Density Lipoprotein and Platelet-Activating Factor-Stimulated Human Monocyte Chemoattractant Protein-1 J. Pharmacol. Exp. Ther., June 1, 2003; 305(3): 846 - 853. [Abstract] [Full Text] [PDF]

				T. Wallerath, D. Poleo, H. Li, and U. Forstermann Red wine increases the expression of human endothelial nitric oxide synthase: a mechanism that may contribute to its beneficial cardiovascular effects J. Am. Coll. Cardiol., February 5, 2003; 41(3): 471 - 478. [Abstract] [Full Text] [PDF]

				J. A. Beckman, M. A. Creager, and P. Libby Diabetes and Atherosclerosis: Epidemiology, Pathophysiology, and Management JAMA, May 15, 2002; 287(19): 2570 - 2581. [Abstract] [Full Text] [PDF]

				S. Stork, K. Baumann, C. von Schacky, and P. Angerer The effect of 17{beta}-estradiol on MCP-1 serum levels in postmenopausal women Cardiovasc Res, February 15, 2002; 53(3): 642 - 649. [Abstract] [Full Text] [PDF]

				J. Pfeilschifter, R. Koditz, M. Pfohl, and H. Schatz Changes in Proinflammatory Cytokine Activity after Menopause Endocr. Rev., February 1, 2002; 23(1): 90 - 119. [Abstract] [Full Text] [PDF]

				A. Godecke, M. Ziegler, Z. Ding, and J. Schrader Endothelial dysfunction of coronary resistance vessels in apoE-/- mice involves NO but not prostacyclin-dependent mechanisms Cardiovasc Res, January 1, 2002; 53(1): 253 - 262. [Abstract] [Full Text] [PDF]

				O. Herkert, H. Kuhl, J. Sandow, R. Busse, and V. B. Schini-Kerth Sex Steroids Used in Hormonal Treatment Increase Vascular Procoagulant Activity by Inducing Thrombin Receptor (PAR-1) Expression: Role of the Glucocorticoid Receptor Circulation, December 4, 2001; 104(23): 2826 - 2831. [Abstract] [Full Text] [PDF]

				B.S. Wung, J.J. Cheng, S.-K. Shyue, and D.L. Wang NO Modulates Monocyte Chemotactic Protein-1 Expression in Endothelial Cells Under Cyclic Strain Arterioscler Thromb Vasc Biol, December 1, 2001; 21(12): 1941 - 1947. [Abstract] [Full Text] [PDF]

				K. K. Koh, M. H. Kang, D. K. Jin, S.-K. Lee, J. Y. Ahn, H. Y. Hwang, S. H. Yang, D. S. Kim, T. H. Ahn, and E. K. Shin Vascular effects of estrogen in type II diabetic postmenopausal women J. Am. Coll. Cardiol., November 1, 2001; 38(5): 1409 - 1415. [Abstract] [Full Text] [PDF]

				T. K. Hsiai, S. K. Cho, S. Reddy, S. Hama, M. Navab, L. L. Demer, H. M. Honda, and C. M. Ho Pulsatile Flow Regulates Monocyte Adhesion to Oxidized Lipid-Induced Endothelial Cells Arterioscler Thromb Vasc Biol, November 1, 2001; 21(11): 1770 - 1776. [Abstract] [Full Text] [PDF]

				A. H. WAGNER, M. R. SCHROETER, and M. HECKER 17{beta}-Estradiol inhibition of NADPH oxidase expression in human endothelial cells FASEB J, October 1, 2001; 15(12): 2121 - 2130. [Abstract] [Full Text] [PDF]

				K. Wang, Z. Zhou, X. Zhou, K. Tarakji, E. J. Topol, and A. M. Lincoff Prevention of intimal hyperplasia with recombinant soluble P-selectin glycoprotein ligand-immunoglobulin in the porcine coronary artery balloon injury model J. Am. Coll. Cardiol., August 1, 2001; 38(2): 577 - 582. [Abstract] [Full Text] [PDF]

				A. Tedgui and Z. Mallat Anti-Inflammatory Mechanisms in the Vascular Wall Circ. Res., May 11, 2001; 88(9): 877 - 887. [Abstract] [Full Text] [PDF]

				V. W. M. van Hinsbergh NO or H2O2 for Endothelium-Dependent Vasorelaxation : Tetrahydrobiopterin Makes the Difference Arterioscler Thromb Vasc Biol, May 1, 2001; 21(5): 719 - 721. [Full Text] [PDF]

				K. K. Koh, D. K. Jin, S. H. Yang, S.-K. Lee, H. Y. Hwang, M. H. Kang, W. Kim, D. S. Kim, I. S. Choi, and E. K. Shin Vascular Effects of Synthetic or Natural Progestagen Combined With Conjugated Equine Estrogen in Healthy Postmenopausal Women Circulation, April 17, 2001; 103(15): 1961 - 1966. [Abstract] [Full Text] [PDF]

				G. Lembo, N. De Luca, C. Battagli, G. Iovino, A. Aretini, M. Musicco, G. Frati, F. Pompeo, C. Vecchione., and B. Trimarco A Common Variant of Endothelial Nitric Oxide Synthase (Glu298Asp) Is an Independent Risk Factor for Carotid Atherosclerosis Stroke, March 1, 2001; 32(3): 735 - 740. [Abstract] [Full Text] [PDF]

				I. Fleming, U. R. Michaelis, D. Bredenkotter, B. Fisslthaler, F. Dehghani, R. P. Brandes, and R. Busse Endothelium-Derived Hyperpolarizing Factor Synthase (Cytochrome P450 2C9) Is a Functionally Significant Source of Reactive Oxygen Species in Coronary Arteries Circ. Res., January 19, 2001; 88(1): 44 - 51. [Abstract] [Full Text] [PDF]

				M. Kroon, P Koolwijk, B van der Vecht, and V. van Hinsbergh Hypoxia in combination with FGF-2 induces tube formation by human microvascular endothelial cells in a fibrin matrix: involvement of at least two signal transduction pathways J. Cell Sci., January 2, 2001; 114(4): 825 - 833. [Abstract] [PDF]

				R. H. Boger, S. M. Bode-Boger, P. S. Tsao, P. S. Lin, J. R. Chan, and J. P. Cooke An endogenous inhibitor of nitric oxide synthase regulates endothelial adhesiveness for monocytes J. Am. Coll. Cardiol., December 1, 2000; 36(7): 2287 - 2295. [Abstract] [Full Text] [PDF]

				M. Koyanagi, K. Egashira, S. Kitamoto, W. Ni, H. Shimokawa, M. Takeya, T. Yoshimura, and A. Takeshita Role of Monocyte Chemoattractant Protein-1 in Cardiovascular Remodeling Induced by Chronic Blockade of Nitric Oxide Synthesis Circulation, October 31, 2000; 102(18): 2243 - 2248. [Abstract] [Full Text] [PDF]

				K. EGASHIRA, M. KOYANAGI, S. KITAMOTO, W. NI, C. KATAOKA, R. MORISHITA, Y. KANEDA, C. AKIYAMA, K.-I. NISHIDA, K. SUEISHI, et al. Anti-monocyte chemoattractant protein-1 gene therapy inhibits vascular remodeling in rats: blockade of MCP-1 activity after intramuscular transfer of a mutant gene inhibits vascular remodeling induced by chronic blockade of NO synthesis FASEB J, October 1, 2000; 14(13): 1974 - 1978. [Abstract] [Full Text]

				M. M. ARRIERO, J. A. RODRÍGUEZ-FEO, A. CELDRÁN, L. S. D. MIGUEL, F. GONZÁLEZ-FERNÁNDEZ, J. FORTES, A. REYERO, O. FRIEYRO, J. C. D. L. PINTA, A. FRANCO, et al. Expression of Endothelial Nitric Oxide Synthase in Human Peritoneal Tissue: Regulation by Escherichia Coli Lipopolysaccharide J. Am. Soc. Nephrol., October 1, 2000; 11(10): 1848 - 1856. [Abstract] [Full Text]

				R. P. Patel, A.-L. Levonen, J. H. Crawford, and V. M. Darley-Usmar Mechanisms of the pro- and anti-oxidant actions of nitric oxide in atherosclerosis Cardiovasc Res, August 18, 2000; 47(3): 465 - 474. [Abstract] [Full Text] [PDF]

				S. Kitamoto, K. Egashira, C. Kataoka, M. Koyanagi, M. Katoh, H. Shimokawa, R. Morishita, Y. Kaneda, K. Sueishi, and A. Takeshita Increased Activity of Nuclear Factor-{kappa}B Participates in Cardiovascular Remodeling Induced by Chronic Inhibition of Nitric Oxide Synthesis in Rats Circulation, August 15, 2000; 102(7): 806 - 812. [Abstract] [Full Text] [PDF]

				K. M. Channon, H. Qian, and S. E. George Nitric Oxide Synthase in Atherosclerosis and Vascular Injury : Insights From Experimental Gene Therapy Arterioscler Thromb Vasc Biol, August 1, 2000; 20(8): 1873 - 1881. [Abstract] [Full Text] [PDF]

				M. J. Somers, K. Mavromatis, Z. S. Galis, and D. G. Harrison Vascular Superoxide Production and Vasomotor Function in Hypertension Induced by Deoxycorticosterone Acetate-Salt Circulation, April 11, 2000; 101(14): 1722 - 1728. [Abstract] [Full Text] [PDF]

				H. Li and U. Förstermann Structure-Activity Relationship of Staurosporine Analogs in Regulating Expression of Endothelial Nitric-Oxide Synthase Gene Mol. Pharmacol., March 1, 2000; 57(3): 427 - 435. [Abstract] [Full Text]

				A. Blum, L. Hathaway, R. Mincemoyer, W. H. Schenke, M. Kirby, G. Csako, M. A. Waclawiw, J. A. Panza, and R. O. Cannon III Effects of oral L-arginine on endothelium-dependent vasodilation and markers of inflammation in healthy postmenopausal women J. Am. Coll. Cardiol., February 1, 2000; 35(2): 271 - 276. [Abstract] [Full Text] [PDF]

				C. Urbich, M. Fritzenwanger, A. M. Zeiher, and S. Dimmeler Laminar Shear Stress Upregulates the Complement-Inhibitory Protein Clusterin : A Novel Potent Defense Mechanism Against Complement-Induced Endothelial Cell Activation Circulation, February 1, 2000; 101(4): 352 - 355. [Abstract] [Full Text] [PDF]

				M. Usui, K. Egashira, H. Tomita, M. Koyanagi, M. Katoh, H. Shimokawa, M. Takeya, T. Yoshimura, K. Matsushima, and A. Takeshita Important Role of Local Angiotensin II Activity Mediated via Type 1 Receptor in the Pathogenesis of Cardiovascular Inflammatory Changes Induced by Chronic Blockade of Nitric Oxide Synthesis in Rats Circulation, January 25, 2000; 101(3): 305 - 310. [Abstract] [Full Text] [PDF]

				J. Niebauer, S. P. Schwarzacher, M. Hayase, B. Wang, R. S. Kernoff, J. P. Cooke, and A. C. Yeung Local L-Arginine Delivery After Balloon Angioplasty Reduces Monocyte Binding and Induces Apoptosis Circulation, October 26, 1999; 100(17): 1830 - 1835. [Abstract] [Full Text] [PDF]

				C. Kunsch and R. M. Medford Oxidative Stress as a Regulator of Gene Expression in the Vasculature Circ. Res., October 15, 1999; 85(8): 753 - 766. [Abstract] [Full Text] [PDF]

				J. Niebauer, J.o. Dulak, J. R. Chan, P. S. Tsao, and J. P. Cooke Gene transfer of nitric oxide synthase: Effects on endothelial biology J. Am. Coll. Cardiol., October 1, 1999; 34(4): 1201 - 1207. [Abstract] [Full Text] [PDF]

				T. de Frutos, L. S. de Miguel, M. Garcia-Duran, F. Gonzalez-Fernandez, J. A. Rodriguez-Feo, M. Monton, J. Guerra, J. Farre, S. Casado, and A. Lopez-Farre NO from smooth muscle cells decreases NOS expression in endothelial cells: role of TNF-alpha Am J Physiol Heart Circ Physiol, October 1, 1999; 277(4): H1317 - H1325. [Abstract] [Full Text] [PDF]

				K. R. WALLEY, T. E. MCDONALD, Y. HIGASHIMOTO, and S. HAYASHI Modulation of Proinflammatory Cytokines by Nitric Oxide in Murine Acute Lung Injury Am. J. Respir. Crit. Care Med., August 1, 1999; 160(2): 698 - 704. [Abstract] [Full Text]

				A. Bellocq, E. Azoulay, S. Marullo, A. Flahault, B. Fouqueray, C. Philippe, J. Cadranel, and L. Baud Reactive Oxygen and Nitrogen Intermediates Increase Transforming Growth Factor-beta 1 Release from Human Epithelial Alveolar Cells through Two Different Mechanisms Am. J. Respir. Cell Mol. Biol., July 1, 1999; 21(1): 128 - 136. [Abstract] [Full Text]

				H. Qian, V. Neplioueva, G. A. Shetty, K. M. Channon, and S. E. George Nitric Oxide Synthase Gene Therapy Rapidly Reduces Adhesion Molecule Expression and Inflammatory Cell Infiltration in Carotid Arteries of Cholesterol-Fed Rabbits Circulation, June 15, 1999; 99(23): 2979 - 2982. [Abstract] [Full Text] [PDF]

				T.-c. Hsieh, G. Juan, Z. Darzynkiewicz, and J. M. Wu Resveratrol Increases Nitric Oxide Synthase, Induces Accumulation of p53 and p21WAF1/CIP1, and Suppresses Cultured Bovine Pulmonary Artery EndothelialCell Proliferation by Perturbing Progression through S and G2 Cancer Res., June 1, 1999; 59(11): 2596 - 2601. [Abstract] [Full Text] [PDF]

				Y.-T. Xuan, X.-L. Tang, S. Banerjee, H. Takano, R. C. X. Li, H. Han, Y. Qiu, J.-J. Li, and R. Bolli Nuclear Factor-{kappa}B Plays an Essential Role in the Late Phase of Ischemic Preconditioning in Conscious Rabbits Circ. Res., May 14, 1999; 84(9): 1095 - 1109. [Abstract] [Full Text] [PDF]

				X. Bao, C. Lu, and J. A. Frangos Temporal Gradient in Shear But Not Steady Shear Stress Induces PDGF-A and MCP-1 Expression in Endothelial Cells : Role of NO, NF{kappa}B, and egr-1 Arterioscler Thromb Vasc Biol, April 1, 1999; 19(4): 996 - 1003. [Abstract] [Full Text] [PDF]

				M.A Barbacanne, J Rami, J.B Michel, J.P Souchard, M Philippe, J.P Besombes, F Bayard, and J.F Arnal Estradiol increases rat aorta endothelium-derived relaxing factor (EDRF) activity without changes in endothelial NO synthase gene expression: possible role of decreased endothelium-derived superoxide anion production Cardiovasc Res, March 1, 1999; 41(3): 672 - 681. [Abstract] [Full Text] [PDF]

				K. Beck, W Eberhardt, S Frank, A Huwiler, U. Messmer, H Muhl, and J Pfeilschifter Inducible NO synthase: role in cellular signalling J. Exp. Biol., January 3, 1999; 202(6): 645 - 653. [Abstract] [PDF]

				M F NEURATH, C BECKER, and K BARBULESCU Role of NF-kappa B in immune and inflammatory responses in the gut Gut, December 1, 1998; 43(6): 856 - 860. [Abstract] [Full Text] [PDF]

				M. A. Hernandez-Presa, C. Bustos, M. Ortego, J. Tunon, L. Ortega, and J. Egido ACE Inhibitor Quinapril Reduces the Arterial Expression of NF-{kappa}B-Dependent Proinflammatory Factors but not of Collagen I in a Rabbit Model of Atherosclerosis Am. J. Pathol., December 1, 1998; 153(6): 1825 - 1837. [Abstract] [Full Text] [PDF]

				C. M. Hogaboam, C. S. Gallinat, C. Bone-Larson, S. W. Chensue, N. W. Lukacs, R. M. Strieter, and S. L. Kunkel Collagen Deposition in a Non-Fibrotic Lung Granuloma Model after Nitric Oxide Inhibition Am. J. Pathol., December 1, 1998; 153(6): 1861 - 1872. [Abstract] [Full Text] [PDF]

				K. M. Channon, H. Qian, V. Neplioueva, M. A. Blazing, E. Olmez, G. A. Shetty, S. A. Youngblood, J. Pawloski, T. McMahon, J. S. Stamler, et al. In Vivo Gene Transfer of Nitric Oxide Synthase Enhances Vasomotor Function in Carotid Arteries From Normal and Cholesterol-Fed Rabbits Circulation, November 3, 1998; 98(18): 1905 - 1911. [Abstract] [Full Text] [PDF]

				X.-L. Chen, P. E. Tummala, M. T. Olbrych, R. W. Alexander, and R. M. Medford Angiotensin II Induces Monocyte Chemoattractant Protein-1 Gene Expression in Rat Vascular Smooth Muscle Cells Circ. Res., November 2, 1998; 83(9): 952 - 959. [Abstract] [Full Text] [PDF]

				M. E. Ritchie Nuclear Factor-{kappa}B Is Selectively and Markedly Activated in Humans With Unstable Angina Pectoris Circulation, October 27, 1998; 98(17): 1707 - 1713. [Abstract] [Full Text] [PDF]

				D. Corseaux, T. Le Tourneau, I. Six, M. D. Ezekowitz, E. P. Mc Fadden, T. Meurice, P. Asseman, C. Bauters, and B. Jude Enhanced Monocyte Tissue Factor Response After Experimental Balloon Angioplasty in Hypercholesterolemic Rabbit: Inhibition With Dietary L-Arginine Circulation, October 27, 1998; 98(17): 1776 - 1782. [Abstract] [Full Text] [PDF]

				M. Katoh, K. Egashira, M. Usui, T. Ichiki, H. Tomita, H. Shimokawa, H. Rakugi, and A. Takeshita Cardiac Angiotensin II Receptors Are Upregulated by Long-Term Inhibition of Nitric Oxide Synthesis in Rats Circ. Res., October 5, 1998; 83(7): 743 - 751. [Abstract] [Full Text] [PDF]

				S. Pervin, R. Singh, M. E. Rosenfeld, M. Navab, G. Chaudhuri, and L. Nathan Estradiol Suppresses MCP-1 Expression In Vivo : Implications for Atherosclerosis Arterioscler Thromb Vasc Biol, October 1, 1998; 18(10): 1575 - 1582. [Abstract] [Full Text] [PDF]

				H. Tomita, K. Egashira, M. Kubo-Inoue, M. Usui, M. Koyanagi, H. Shimokawa, M. Takeya, T. Yoshimura, and A. Takeshita Inhibition of NO Synthesis Induces Inflammatory Changes and Monocyte Chemoattractant Protein-1 Expression in Rat Hearts and Vessels Arterioscler Thromb Vasc Biol, September 1, 1998; 18(9): 1456 - 1464. [Abstract] [Full Text] [PDF]

				H. Tomita, K. Egashira, Y. Ohara, M. Takemoto, M. Koyanagi, M. Katoh, H. Yamamoto, K. Tamaki, H. Shimokawa, and A. Takeshita Early Induction of Transforming Growth Factor-ß via Angiotensin II Type 1 Receptors Contributes to Cardiac Fibrosis Induced by Long-term Blockade of Nitric Oxide Synthesis in Rats Hypertension, August 1, 1998; 32(2): 273 - 279. [Abstract] [Full Text] [PDF]

				S. Ravalli, A. Albala, M. Ming, M. Szabolcs, A. Barbone, R. E. Michler, and P. J. Cannon Inducible Nitric Oxide Synthase Expression in Smooth Muscle Cells and Macrophages of Human Transplant Coronary Artery Disease Circulation, June 16, 1998; 97(23): 2338 - 2345. [Abstract] [Full Text] [PDF]

				P. S. Tsao, J. Niebauer, R. Buitrago, P. S. Lin, B.-y. Wang, J. P. Cooke, Y-d. Ida Chen, and G. M. Reaven Interaction of Diabetes and Hypertension on Determinants of Endothelial Adhesiveness Arterioscler Thromb Vasc Biol, June 1, 1998; 18(6): 947 - 953. [Abstract] [Full Text] [PDF]

				D. Sekkai, F. Aillet, N. Israel, and M. Lepoivre Inhibition of NF-kappa B and HIV-1 Long Terminal Repeat Transcriptional Activation by Inducible Nitric Oxide Synthase 2 Activity J. Biol. Chem., February 13, 1998; 273(7): 3895 - 3900. [Abstract] [Full Text] [PDF]

				C. M. Hogaboam, M. L. Steinhauser, H. Schock, N. Lukacs, R. M. Strieter, T. Standiford, and S. L. Kunkel Therapeutic Effects of Nitric Oxide Inhibition during Experimental Fecal Peritonitis: Role of Interleukin-10 and Monocyte Chemoattractant Protein 1 Infect. Immun., February 1, 1998; 66(2): 650 - 655. [Abstract] [Full Text] [PDF]

				J. S. Luoma, P. Stralin, S. L. Marklund, T. P. Hiltunen, T. Sarkioja, and S. Yla-Herttuala Expression of Extracellular SOD and iNOS in Macrophages and Smooth Muscle Cells in Human and Rabbit Atherosclerotic Lesions : Colocalization With Epitopes Characteristic of Oxidized LDL and Peroxynitrite-Modified Proteins Arterioscler Thromb Vasc Biol, February 1, 1998; 18(2): 157 - 167. [Abstract] [Full Text] [PDF]

				C. Hermann, A. M. Zeiher, and S. Dimmeler Shear Stress Inhibits H2O2-Induced Apoptosis of Human Endothelial Cells by Modulation of the Glutathione Redox Cycle and Nitric Oxide Synthase Arterioscler Thromb Vasc Biol, December 1, 1997; 17(12): 3588 - 3592. [Abstract] [Full Text]

				V. Guetta, A. A. Quyyumi, A. Prasad, J. A. Panza, M. Waclawiw, and R. O. Cannon III The Role of Nitric Oxide in Coronary Vascular Effects of Estrogen in Postmenopausal Women Circulation, November 4, 1997; 96(9): 2795 - 2801. [Abstract] [Full Text]

				P. Skarsgard, C. Van Breemen, and I. Laher Estrogen regulates myogenic tone in pressurized cerebral arteries by enhanced basal release of nitric oxide Am J Physiol Heart Circ Physiol, November 1, 1997; 273(5): H2248 - H2256. [Abstract] [Full Text] [PDF]

				T. Marumo, V. B. Schini-Kerth, B. Fisslthaler, and R. Busse Platelet-Derived Growth Factor–Stimulated Superoxide Anion Production Modulates Activation of Transcription Factor NF-{kappa}B and Expression of Monocyte Chemoattractant Protein 1 in Human Aortic Smooth Muscle Cells Circulation, October 7, 1997; 96(7): 2361 - 2367. [Abstract] [Full Text]

				J.-L. Balligand and P. J. Cannon Nitric Oxide Synthases and Cardiac Muscle : Autocrine and Paracrine Influences Arterioscler Thromb Vasc Biol, October 1, 1997; 17(10): 1846 - 1858. [Abstract] [Full Text]

				M. J. Thomassen, L. T. Buhrow, M. J. Connors, F. Kaneko, S. C. Erzurum, and M. S. Kavuru Nitric Oxide Inhibits Inflammatory Cytokine Production by Human Alveolar Macrophages Am. J. Respir. Cell Mol. Biol., September 1, 1997; 17(3): 279 - 283. [Abstract] [Full Text]

				S. P. Schwarzacher, T. T. Lim, B. Wang, R. S. Kernoff, J. Niebauer, J. P. Cooke, and A. C. Yeung Local Intramural Delivery of L-Arginine Enhances Nitric Oxide Generation and Inhibits Lesion Formation After Balloon Angioplasty Circulation, April 1, 1997; 95(7): 1863 - 1869. [Abstract] [Full Text]

				M. Hernandez-Presa, C. Bustos, M. Ortego, J. Tunon, G. Renedo, M. Ruiz-Ortega, and J. Egido Angiotensin-Converting Enzyme Inhibition Prevents Arterial Nuclear Factor-{kappa}B Activation, Monocyte Chemoattractant Protein-1 Expression, and Macrophage Infiltration in a Rabbit Model of Early Accelerated Atherosclerosis Circulation, March 18, 1997; 95(6): 1532 - 1541. [Abstract] [Full Text]

				M. R. Adams, W. Jessup, D. Hailstones, and D. S. Celermajer L-Arginine Reduces Human Monocyte Adhesion to Vascular Endothelium and Endothelial Expression of Cell Adhesion Molecules Circulation, February 4, 1997; 95(3): 662 - 668. [Abstract] [Full Text]

				E. Allaire and A. W. Clowes Endothelial Cell Injury in Cardiovascular Surgery: The Intimal Hyperplastic Response Ann. Thorac. Surg., February 1, 1997; 63(2): 582 - 591. [Abstract] [Full Text]

				J. P. Cooke and P. S. Tsao Arginine: A New Therapy for Atherosclerosis? Circulation, January 21, 1997; 95(2): 311 - 312. [Full Text]

				W. Aji, S. Ravalli, M. Szabolcs, X.-c. Jiang, R. R. Sciacca, R. E. Michler, and P. J. Cannon L-Arginine Prevents Xanthoma Development and Inhibits Atherosclerosis in LDL Receptor Knockout Mice Circulation, January 21, 1997; 95(2): 430 - 437. [Abstract] [Full Text]

				W. Schaper and W. D. Ito Molecular Mechanisms of Coronary Collateral Vessel Growth Circ. Res., November 1, 1996; 79(5): 911 - 919. [Full Text]

				P. S. Tsao, R. Buitrago, J. R. Chan, and J. P. Cooke Fluid Flow Inhibits Endothelial Adhesiveness: Nitric Oxide and Transcriptional Regulation of VCAM-1 Circulation, October 1, 1996; 94(7): 1682 - 1689. [Abstract] [Full Text]

				R. C. Candipan, B.-y. Wang, R. Buitrago, P. S. Tsao, and J. P. Cooke Regression or Progression : Dependency on Vascular Nitric Oxide Arterioscler Thromb Vasc Biol, January 1, 1996; 16(1): 44 - 50. [Abstract] [Full Text]

This Article Abstract Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Zeiher, A. M. Articles by Busse, R. Search for Related Content PubMed PubMed Citation Articles by Zeiher, A. M. Articles by Busse, R.