AJP - Heart Fuel your research with LabChart
HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
QUICK SEARCH:   [advanced]


     

Am J Physiol Heart Circ Physiol 291: H1694-H1699, 2006; doi:10.1152/ajpheart.00340.2006
0363-6135/06 $8.00
This Article
Right arrow Abstract Freely available
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in ISI Web of Science
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via ISI Web of Science (17)
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Csiszar, A.
Right arrow Articles by Ungvari, Z.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Csiszar, A.
Right arrow Articles by Ungvari, Z.

Resveratrol attenuates TNF-{alpha}-induced activation of coronary arterial endothelial cells: role of NF-{kappa}B inhibition

Anna Csiszar, Kira Smith, Nazar Labinskyy, Zsuzsanna Orosz, Aracelie Rivera, and Zoltan Ungvari

Department of Physiology, New York Medical College, Valhalla, New York

Submitted 30 March 2006 ; accepted in final form 8 May 2006


   ABSTRACT
TOP
 ABSTRACT
MATERIALS AND METHODS
 RESULTS
 DISCUSSION
 GRANTS
 REFERENCES
 
Epidemiological studies suggest that Mediterranean diets rich in resveratrol are associated with reduced risk of coronary artery disease. However, the mechanisms by which resveratrol exerts its cardioprotective effects are not completely understood. Because TNF-{alpha}-induced endothelial activation and vascular inflammation play a critical role in vascular aging and atherogenesis, we evaluated whether resveratrol inhibits TNF-{alpha}-induced signal transduction in human coronary arterial endothelial cells (HCAECs). We found that TNF-{alpha} significantly increased adhesiveness of the monocytic THP-1 cells to HCAECs, an effect that could be inhibited by pretreatment with resveratrol and the NF-{kappa}B inhibitor pyrrolidine dithiocarbamate. Previously, we found that TNF-{alpha} activates NAD(P)H oxidases, and our recent data showed that TNF-{alpha}-induced endothelial activation was prevented by the NAD(P)H oxidase inhibitor apocynin or catalase plus SOD. Resveratrol also inhibited H2O2-induced monocyte adhesiveness. Using a reporter gene assay, we found that, in HCAECs, TNF-{alpha} significantly increased NF-{kappa}B activity, which could be inhibited by resveratrol (>50% inhibition at 10–6 mol/l) and pyrrolidine dithiocarbamate. Resveratrol also inhibited TNF-{alpha}-induced, NF-{kappa}B-driven luciferase expression in rat aortas electroporated with the reporter gene construct. In TNF-{alpha}-treated HCAECs, resveratrol (in the submicromolar range) significantly attenuated expression of NF-{kappa}B-dependent inflammatory markers inducible nitric oxide synthase, IL-6, bone morphogenetic protein-2, ICAM-1, and VCAM. Thus resveratrol at nutritionally relevant concentrations inhibits TNF-{alpha}-induced NF-{kappa}B activation and inflammatory gene expression and attenuates monocyte adhesiveness to HCAECs. We propose that these anti-inflammatory actions of resveratrol are responsible, at least in part, for its cardioprotective effects.

stilbene; atheroprotection; phytoestrogen; vascular aging; inflammation

PROINFLAMMATORY CYTOKINES, including TNF-{alpha}, are thought to play an important role in vascular inflammation, leading to atherosclerosis and development of coronary arterial disease (2, 18–20, 26, 29, 31, 38). There is ample evidence that TNF-{alpha} and other proinflammatory cytokines elicit NF-{kappa}B activation in endothelial cells (24, 40), which mediates, at least in part, their proatherogenic effects. These include induction of chemokines and adhesion molecules [e.g., VCAM, ICAM-1 (40)], which promote monocyte adhesiveness to the endothelium and upregulation of factors involved in plaque development [e.g., IL-6, bone morphogenetic proteins (4, 7)]. There is growing evidence that disruption of the cytokine-induced NF-{kappa}B signaling pathway exerts significant vasculoprotective action by attenuating vascular inflammation (22, 23, 30), which delays or prevents atherogenesis in animal disease models (16). Thus identifying novel pharmacological methods to inhibit vascular inflammation and NF-{kappa}B activation in coronary arteries holds great promise for the prevention of ischemic heart disease in humans (3, 22).

Epidemiological studies suggest that Mediterranean diets are associated with reduced risk of cardiovascular disease (12, 17). It has been proposed that resveratrol is an important dietary constituent involved in vasculoprotection. Resveratrol has been identified in more than 70 species of plants, including grapevines (Vitis vinifera), mulberries, and peanuts, and it is thought to have diverse antiatherogenic activities (34–36, 43, 44), such as the inhibition of LDL oxidation, platelet aggregation, and regulation of vascular smooth muscle proliferation. It is significant that previous studies on myeloid (U-937), lymphoid (Jurkat), and epithelial (HeLa and H4) cell lines raised the possibility that resveratrol may interfere with NF-{kappa}B signaling (21, 27). Yet the effects of resveratrol on cytokine-induced NF-{kappa}B activation, upregulation of inflammatory mediators, and monocyte adhesion in coronary arterial endothelial cells are less understood. Thus in the present study we aimed to elucidate whether, in human coronary arterial endothelial cells (HCAECs), resveratrol can inhibit TNF-{alpha}-induced NF-{kappa}B activation and monocyte adhesiveness.


   MATERIALS AND METHODS
TOP
 ABSTRACT
 MATERIALS AND METHODS
RESULTS
 DISCUSSION
 GRANTS
 REFERENCES
 
Cell cultures. Primary HCAECs (Cell Applications) were cultured as described (4, 7).

Monocyte adhesion assay. We measured adhesion of fluorescently labeled human monocytic (THP-1) cells to confluent monolayers of HCAECs using a microplate-based assay. In brief, HCAECs were grown to confluence in 96-well plates and were treated with TNF-{alpha} or IL-6 (0.1–10 ng/ml; incubation time: 2 h, at 37°C) in the absence or presence (60-min preincubation) of resveratrol (0.1–100 µmol/l) or pyrrolidine dithiocarbamate (10 µmol/l). THP-1 cells were labeled with the fluorescent dye calcein (5 µmol/l final concentration; Molecular Probes, Eugene, OR; in serum-free RPMI medium for 30 min at 37°C). Then cells were washed twice with prewarmed (37°C) RPMI. Phorbol myristate acetate (10–6 mol/l)-pretreated fluorescently labeled THP-1 cells (5 x 105/well) were added to the microplate wells containing confluent HCAECs (medium removed; incubation time: 120 min, at 37°C). Nonadherent THP-1 cells were removed by careful washing (three times with prewarmed RPMI). Then 200 µl of PBS were added to each well, and fluorescence was measured using an Flx-800 (Bio-Tek Instruments) fluorescent plate reader (excitation: 485 nm; emission: 528 nm). Controls included measurement of total fluorescence of labeled cells before adhesion, controls for measuring autofluorescence of unlabeled cells, and measurement of monocyte adhesion to endothelial cell-free microplate wells.

Transient transfection and luciferase assays. Effect of TNF-{alpha} on NF-{kappa}B activity in HCAECs was tested by a reporter gene assay as described (4, 5, 7). We used a NF-{kappa}B reporter composed of an NF-{kappa}B response element upstream of firefly luciferase (NF-{kappa}B-Luc, Stratagene) and a renilla luciferase plasmid under the control of the cytomegalovirus (CMV) promoter (as an internal control). Transfections in HCAECs were performed using the Amaxa Nucleofector technology (Amaxa, Gaithersburg, MD), as our laboratory has previously reported (4, 5, 7). Firefly and renilla luciferase activities were assessed after 24 h using the Dual Luciferase Reporter Assay Kit (Promega) and a luminometer.

Vessel culture and conditions of electroporation. All animal protocols were approved by the Institutional Animal Care and Use Committee of the New York Medical College, Valhalla, NY. Aortic segments were isolated from male Wistar rats (weighing 150 g) and cleaned from the surrounding tissues under sterile conditions. Square-wave electric pulses were delivered to the vessels with a cylindrical external electrode and an intraluminal electrode (1 cm long, 1-mm fixed distance between the electrodes; see GoGoFig. 3C) by using an electric pulse generator (model CUY 201 BTX; Protech International, San Antonio, TX), and then the vessel segments were maintained in organoid culture for 24 h as described (4, 7). The electric pulse was regulated as follows: voltage, 20 V; pulse-on time, 10 ms; interval time, 990 ms; and number of pulses, 10. These optimized parameters were determined in preliminary studies by measuring luciferase activity 1 day after electroporation of a CMV-driven renilla luciferase construct at various electrode voltages, pulse numbers, and pulse durations, according to the modified protocols of Matsumoto et al. (25) and Yamaoka et al. (41). In separate experiments, electroporation with a pDsRed-Monomer vector (Clontech, Mountain View, CA) that expresses the red fluorescent DsRed-monomer fluorescent protein was used to assess the efficiency of the endothelial transfection by this method. Immunolabeling for {alpha}-smooth muscle actin (for the methods, see Ref. 32) was used to confirm the integrity of the vessel wall. Viability of endothelial and smooth muscle cells was assessed by measuring the magnitude of phenylephrine-induced contractions and acetylcholine-induced relaxations of vascular ring preparations, as described (4). The concentration of the plasmid DNA solution used for transfection was adjusted to 1.5 µg/µl. After overnight culture, some vessels were pretreated with resveratrol (10 µmol/l, for 2 h). Then the aortas were incubated with TNF-{alpha} (10 ng/ml, for 4 h) and homogenized in 500 µl of dual luciferase assay lysis reagent (Promega). The firefly and renilla luciferase activities in homogenates of transfected aortas were measured by a procedure described previously (7).


Figure 1
View larger version (36K):
[in this window]
[in a new window]
  		Fig. 1. Results of monocyte adhesion assay (see MATERIALS AND METHODS). Treatment (2 h) of primary human coronary arterial endothelial cells (HCAECs) with increasing concentrations of TNF-{alpha} (A) and IL-6 (B) significantly increased the adhesion of fluorescently labeled phorbol myristate acetate-stimulated THP-1 monocytic cells. The effects of 10 ng/ml TNF-{alpha} and IL-6 were also assessed after pretreatment with increasing concentrations of resveratrol or an inhibitor of NF-{kappa}B [10 µmol/l pyrrolidine dithiocarbamate (PDTC)]. Values are means ± SE (n = 8 for each group). *P < 0.05 vs. untreated control. #P < 0.05 vs. cytokine treatment alone.

 

Figure 2
View larger version (28K):
[in this window]
[in a new window]
  		Fig. 2. Results of monocyte adhesion assay (see MATERIALS AND METHODS). A: pretreatment of primary HCAECs with apocynin or the free-radical scavenger SOD plus catalase (CAT) significantly inhibited adhesion of fluorescently labeled phorbol myristate acetate-stimulated THP-1 monocytic cells induced by 10 ng/ml TNF-{alpha}. B: treatment of HCAECs with increasing concentrations of H2O2 significantly increased monocyte adhesion, which was inhibited by resveratrol. Values are means ± SE (n = 6–8 for each group). *P < 0.05 vs. untreated control. #P < 0.05 vs. cytokine treatment alone.

 

Figure 3
View larger version (47K):
[in this window]
[in a new window]
  		Fig. 3. A: reporter gene assay showing the effects of TNF-{alpha} (10 ng/ml) on NF-{kappa}B reporter activity in cultured primary HCAECs in the absence and presence of resveratrol. Cells were transiently cotransfected with NF-{kappa}B-driven firefly luciferase and cytomegalovirus-driven renilla luciferase constructs followed by TNF-{alpha} stimulation. Cells were then lysed and subjected to luciferase activity assay. After normalization, relative luciferase activity was obtained from six independent transfections. In control experiments, PDTC was used to inhibit NF-{kappa}B activity. Values are means ± SE. *P < 0.05 vs. control; #P < 0.05 vs. without resveratrol. B: resveratrol inhibits the effect of TNF-{alpha} (10 ng/ml) on NF-{kappa}B reporter activity in rat aortas electroporated with the reporter gene construct. Values are means ± SE. *P < 0.05 vs. control; #P < 0.05 vs. without resveratrol. C: schematic representation of the electroporation method for the transfer of genes into the vascular endothelium. Luminal space of the vessel segments was filled with DNA solution, and electric pulses were delivered to the vessel wall using an intraluminal electrode. D, left: representative fluorescent image showing the endothelial expression of DsRed (red fluorescence; arrows) in a rat aorta segment electroporated with the pDsRed-Monomer Vector (see MATERIALS AND METHODS). Middle: overlay of red fluorescent image and bright field image. Right: immunolabeling for {alpha}-smooth muscle actin (green fluorescence) shows that there is no tissue necrosis in the electroporated vessel. 4',6-Diamidino-2-phenylindole dihydrochloride (DAPI; blue) was used for nuclear staining. E: acetylcholine-induced relaxation in cultured (24 h) segments of the same rat aorta electroporated with different voltage settings. F: reporter gene assay showing the effects of resveratrol on IL-1beta and IL-6 (10 ng/ml)-induced NF-{kappa}B reporter activity in HCAECs. PE, phenylephrine. Values are means ± SE. *P < 0.05 vs. control; #P < 0.05 vs. without resveratrol.

 
Quantitative real-time PCR. Total RNA from HCAECs was isolated with Mini RNA Isolation Kit (Zymo Research, Orange, CA) and was reverse transcribed using Superscript III RT (Invitrogen), as described previously (8, 9). Real-time RT-PCR technique was used to analyze mRNA expression using the Strategen MX3000, as reported (4, 7). Efficiency of the PCR reaction was determined using dilution series of a standard vascular sample. Quantification was performed using the {Delta}{Delta}CT method. The housekeeping gene GAPDH was used for internal normalization. Oligonucleotides used for real-time quantitative RT-PCR are listed in Table 1. Fidelity of the PCR reaction was determined by melting temperature analysis and visualization of product on a 2% agarose gel.


View this table:
[in this window]
[in a new window]
  		Table 1. Oligonucleotides for real-time RT-PCR

 
Data analysis. Data were normalized to the respective control mean values and are expressed as means ± SE. Statistical analyses of data were performed by Student's t-test or by two-way ANOVA followed by the Tukey post hoc test, as appropriate. P < 0.05 was considered statistically significant.


   RESULTS
TOP
 ABSTRACT
 MATERIALS AND METHODS
 RESULTS
DISCUSSION
 GRANTS
 REFERENCES
 
Resveratrol inhibits TNF-{alpha}, IL-6, and H2O2-induced increases in monocyte adhesiveness to HCAECs. TNF-{alpha} and IL-6 significantly increased monocyte adherence to cultured HCAECs in a concentration-dependent manner (Fig. 1, A and B, respectively). Pretreatment of the vessels with increasing concentrations of resveratrol reduced or prevented monocyte adhesion induced by both cytokines (Fig. 1, A and B). TNF-{alpha}-induced increases in monocyte adhesiveness were significantly reduced by pretreatment of HCAECs with apocynin or SOD plus catalase (Fig. 2A). Administration of H2O2 also substantially increased monocyte adherence to HCAECs in all concentrations studied (Fig. 2B), and this effect was also attenuated by resveratrol (Fig. 2B).

Resveratrol inhibits cytokine-induced NF-{kappa}B activation in HCAECs. To determine the effect of resveratrol on TNF-{alpha}-induced NF-{kappa}B activation, we transiently transfected HCAECs with a NF-{kappa}B-driven reporter gene construct and then pretreated the cells with resveratrol followed by stimulation with TNF-{alpha} (10 ng/ml, for 2 h). A significant increase in luciferase activity over the vector control was noted upon stimulation with TNF-{alpha} in the absence of resveratrol (Fig. 3A). Pretreatment of HCAECs (for 1 h) with resveratrol prevented TNF-{alpha}-induced NF-{kappa}B activation in a concentration-dependent manner (Fig. 3A). IL-1beta and IL-6 also activated NF-{kappa}B in HCAECs, and these effects were also significantly attenuated by resveratrol (Fig. 3E).

Resveratrol inhibits NF-{kappa}B activation in cultured aortas. First, we determined the optimal conditions for electroporation of reporter gene constructs. We found that luciferase activity increased in proportion to the voltage up to 20 V. However, at higher voltages, vascular luciferase activity was decreased. Acetylcholine-induced relaxations were unaltered up to 30 V (Fig. 3E); however, higher voltages resulted in a significant impairment of endothelium-dependent relaxations (not shown). The luciferase activity increased in proportion to pulse-on time and reached its maximum at 10 ms. Using a red fluorescent protein construct, we demonstrated that, using these voltage settings, the vascular endothelium can be effectively transfected (Fig. 3D). In cultured aortic segments transfected with the NF-{kappa}B reporter construct, TNF-{alpha} elicited significant increases in luciferase activity (Fig. 3B). TNF-{alpha}-induced increases in NF-{kappa}B activity were abolished by pretreatment with resveratrol (Fig. 3B).

Resveratrol inhibits cytokine-induced upregulation of inflammatory mediators in HCAECs. In HCAECs, TNF-{alpha} significantly increased mRNA expression of inducible nitric oxide synthase, IL-6, bone morphogenetic protein-2, ICAM-1, and VCAM in a concentration-dependent manner (Fig. 4, A–C). Pretreatment with increasing concentrations of resveratrol attenuated or prevented TNF-{alpha}-induced upregulation of each inflammatory marker (Fig. 4).


Figure 4
View larger version (19K):
[in this window]
[in a new window]
  		Fig. 4. Effect of TNF-{alpha} (10 ng/ml) on the mRNA expression of IL-6 (A), inducible nitric oxide synthase (iNOS; B), bone morphogenetic protein (BMP-2; C), ICAM-1 (D), and VCAM (E) in cultured primary HCAECs. The effects of TNF-{alpha} were also assessed after pretreatment with increasing concentrations of resveratrol. Analysis of mRNA expression was performed by real-time quantitative RT-PCR. GAPDH was used for normalization. Values are means ± SE (n = 3–5 for each group). *P < 0.05 vs. untreated control, #P < 0.05 vs. without resveratrol.

 

   DISCUSSION
TOP
 ABSTRACT
 MATERIALS AND METHODS
 RESULTS
 DISCUSSION
GRANTS
 REFERENCES
 
There are three major findings in this study. First, we have shown that TNF-{alpha}-induced increased monocyte adhesiveness to HCAECs is NF-{kappa}B dependent, and it can be inhibited by resveratrol (Fig. 1A). IL-6 also elicited endothelial activation, and this effect also could be attenuated by resveratrol (Fig. 1B). It is significant that resveratrol also attenuated H2O2-induced monocyte adhesion to HCAECs in a similar concentration range (Fig. 2B). The findings that TNF-{alpha}-induced monocyte adhesiveness was also attenuated both by inhibition of NAD(P)H oxidase and by reactive oxygen species scavengers (Fig. 2A) suggest that NAD(P)H oxidase-derived H2O2 plays a central role in endothelial activation. Indeed, previous studies by us and others demonstrated that TNF-{alpha} activates NAD(P)H oxidase(s) (11, 32) in endothelial cells.

The second important finding is that TNF-{alpha}-induced NF-{kappa}B activation in HCAECs is inhibited by pretreatment with resveratrol (Fig. 3A). We confirmed that resveratrol was effective against TNF-{alpha}-induced NF-{kappa}B activation in intact blood vessels as well (Fig. 3B). Besides TNF-{alpha}, NF-{kappa}B can also be activated by other proinflammatory cytokines in many cell types. Accordingly, we found that IL-6 and IL-1beta also activated NF-{kappa}B in HCAECs and that resveratrol effectively inhibited the activation of NF-{kappa}B induced by both cytokines (Fig. 3E). These results suggest that resveratrol may act at a step in which all of these agents converge in the signal transduction pathway leading to NF-{kappa}B activation. Importantly, we have evidence that administration of H2O2 activates NF-{kappa}B in coronary arterial endothelial cells (5, 7). In line with this finding, previously we demonstrated that TNF-{alpha}-induced endothelial NF-{kappa}B activation can also be prevented by catalase and NAD(P)H inhibitors (5, 7). Taken together, these results suggest that NAD(P)H oxidase-derived H2O2 mediates cytokine-induced activation of NF-{kappa}B and that resveratrol interferes with this process.

The mechanism of action of resveratrol is not completely understood. It seems that resveratrol-induced inhibition of NF-{kappa}B activation is not cell type specific (21, 27). Resveratrol was reported to block the phosphorylation of p65 subunit of NF-{kappa}B inhibiting the nuclear translocation of NF-{kappa}B (21). It is to be seen whether antioxidant action of resveratrol (e.g., scavenging of H2O2) contributes to its inhibitory effects on NF-{kappa}B activation (18). Interestingly, resveratrol does not inhibit TNF-{alpha}-dependent phosphorylation and degradation of I{kappa}B{alpha} (21). Resveratrol is also a putative activator of SIRT1 (15, 37), a nicotinamide adenosine dinucleotide-dependent histone deacetylase, which may regulate the transcriptional activity of NF-{kappa}B (42). Indeed, SIRT1 was reported to physically interact with the RelA/p65 subunit of NF-{kappa}B and to inhibit transcription by deacetylating RelA/p65 (42). Whether SIRT1 plays a role in the endothelial effects of resveratrol is yet to be determined.

The third important finding of this study is that resveratrol pretreatment prevented TNF-{alpha}-induced upregulation of the inflammatory mediators inducible nitric oxide synthase, IL-6, bone morphogenetic protein-2 (4, 7), ICAM-1 (39), and VCAM (Fig. 4). The transcriptional regulation of these molecules is known to involve NF-{kappa}B; thus it is logical to assume that the observed effects are due to the inhibition of NF-{kappa}B activation by resveratrol. Because all of these factors have been implicated in the development of atherosclerotic plaques, anti-NF-{kappa}B activity of resveratrol is likely antiatherogenic.

Taken together, our results and findings by other laboratories (21) indicate that submicromolar levels of resveratrol are sufficient to suppress cytokine-induced, NF-{kappa}B-dependent cellular responses. With the consideration that each gram of fresh grape skin contains 50–100 µg resveratrol and red wines have 1.5–3 mg/l, the resveratrol concentrations used in our studies are achievable in vivo by consumption of grapes, berries, and/or red wine. It is also important to note that many studies showed that other phenols that are likely to exert resveratrol-like biological actions are also present in the Mediterranean diet and red wines in high concentrations (1, 28, 39).

In a series of elegant studies, Dr. Sinclair's group has showed that resveratrol extends longevity in lower organisms (15, 37), and there is good reason to believe that it also exerts antiaging activity in mammals (recently reviewed in Ref. 18). Cardiovascular aging is characterized by upregulation of TNF-{alpha} (6, 8–10, 13, 33) and NF-{kappa}B activation (14) (A. Csiszar and Z. Ungvari, unpublished observations) associated with a proinflammatory shift in endothelial phenotype in coronary arteries (6, 8–10, 33). Thus future studies should elucidate whether anti-NF-{kappa}B action of resveratrol contributes to its antiaging vasculoprotective effects in the elderly (18).


   GRANTS
TOP
 ABSTRACT
 MATERIALS AND METHODS
 RESULTS
 DISCUSSION
 GRANTS
REFERENCES
 
This work was supported by grants from the American Heart Association (0430108N, 0435140N), the National Heart, Lung, and Blood Institute (HL-077256), and by Philip Morris USA and Philip Morris International.


   FOOTNOTES
 

Address for reprint requests and other correspondence: A. Csiszar, Dept. of Physiology, New York Medical College, Valhalla, NY 10595 (e-mail: anna_csiszar{at}nymc.edu)

The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.


   REFERENCES
TOP
 ABSTRACT
 MATERIALS AND METHODS
 RESULTS
 DISCUSSION
 GRANTS
 REFERENCES
 

  1. Ashikawa K, Majumdar S, Banerjee S, Bharti AC, Shishodia S, and Aggarwal BB. Piceatannol inhibits TNF-induced NF-kappaB activation and NF-kappaB-mediated gene expression through suppression of IkappaBalpha kinase and p65 phosphorylation. J Immunol 169: 6490–6497, 2002.[Abstract/Free Full Text]
  2. Branen L, Hovgaard L, Nitulescu M, Bengtsson E, Nilsson J, and Jovinge S. Inhibition of tumor necrosis factor-alpha reduces atherosclerosis in apolipoprotein E knockout mice. Arterioscler Thromb Vasc Biol 24: 2137–2142, 2004.[Abstract/Free Full Text]
  3. Chen YX, Ma X, Whitman S, and O'Brien ER. Novel antiinflammatory vascular benefits of systemic and stent-based delivery of ethylisopropylamiloride. Circulation 110: 3721–3726, 2004.[Abstract/Free Full Text]
  4. Csiszar A, Ahmad M, Smith KE, Labinskyy N, Gao Q, Kaley G, Edwards JG, Wolin MS, and Ungvari Z. Bone morphogenetic protein-2 induces proinflammatory endothelial phenotype. Am J Pathol 168: 629–638, 2006.[Abstract/Free Full Text]
  5. Csiszar A, Ahmad M, Smith KE, Labinskyy N, Gao Q, Kaley G, Edwards JG, Wolin MS, and Ungvari Z. Bone morphogenetic protein-2 induces proinflammatory endothelial phenotype. Am J Pathol 168: 629–638, 2006.[Abstract/Free Full Text]
  6. Csiszar A, Pacher P, Kaley G, and Ungvari Z. Role of oxidative and nitrosative stress, longevity genes and poly(ADP-ribose) polymerase in cardiovascular dysfunction associated with aging. Curr Vasc Pharmacol 3: 285–291, 2005.[CrossRef][Medline]
  7. Csiszar A, Smith KE, Koller A, Kaley G, Edwards JG, and Ungvari Z. Regulation of bone morphogenetic protein-2 expression in endothelial cells: role of nuclear factor-kappaB activation by tumor necrosis factor-alpha, H2O2, and high intravascular pressure. Circulation 111: 2364–2372, 2005.[Abstract/Free Full Text]
  8. Csiszar A, Ungvari Z, Edwards JG, Kaminski PM, Wolin MS, Koller A, and Kaley G. Aging-induced phenotypic changes and oxidative stress impair coronary arteriolar function. Circ Res 90: 1159–1166, 2002.[Abstract/Free Full Text]
  9. Csiszar A, Ungvari Z, Koller A, Edwards JG, and Kaley G. Aging-induced proinflammatory shift in cytokine expression profile in rat coronary arteries. FASEB J 17: 1183–1185, 2003.[Abstract/Free Full Text]
  10. Csiszar A, Ungvari Z, Koller A, Edwards JG, and Kaley G. Proinflammatory phenotype of coronary arteries promotes endothelial apoptosis in aging. Physiol Genomics 17: 21–30, 2004.[Abstract/Free Full Text]
  11. De Keulenaer GW, Alexander RW, Ushio-Fukai M, Ishizaka N, and Griendling KK. Tumour necrosis factor alpha activates a p22phox-based NADH oxidase in vascular smooth muscle. Biochem J 329: 653–657, 1998.[ISI][Medline]
  12. De Lorgeril M, Salen P, Martin JL, Monjaud I, Delaye J, and Mamelle N. Mediterranean diet, traditional risk factors, and the rate of cardiovascular complications after myocardial infarction: final report of the Lyon Diet Heart Study. Circulation 99: 779–785, 1999.[Abstract/Free Full Text]
  13. Edwards MG, Sarkar D, Klopp R, Morrow JD, Weindruch R, and Prolla TA. Age-related impairment of the transcriptional responses to oxidative stress in the mouse heart. Physiol Genomics 13: 119–127, 2003.[Abstract/Free Full Text]
  14. Helenius M, Hanninen M, Lehtinen SK, and Salminen A. Aging-induced up-regulation of nuclear binding activities of oxidative stress responsive NF-{kappa}B transcription factor in mouse cardiac muscle. J Mol Cell Cardiol 28: 487–498, 1996.[CrossRef][ISI][Medline]
  15. Howitz KT, Bitterman KJ, Cohen HY, Lamming DW, Lavu S, Wood JG, Zipkin RE, Chung P, Kisielewski A, Zhang LL, Scherer B, and Sinclair DA. Small molecule activators of sirtuins extend Saccharomyces cerevisiae lifespan. Nature 425: 191–196, 2003.[CrossRef][Medline]
  16. Jawien J, Gajda M, Mateuszuk L, Olszanecki R, Jakubowski A, Szlachcic A, Korabiowska M, and Korbut R. Inhibition of nuclear factor-kappaB attenuates artherosclerosis in apoE/LDLR-double knockout mice. J Physiol Pharmacol 56: 483–489, 2005.[ISI][Medline]
  17. Keys A, Menotti A, Karvonen MJ, Aravanis C, Blackburn H, Buzina R, Djordjevic BS, Dontas AS, Fidanza F, Keys MH, Kromhout D, Nedeljkovic SO, Punsar S, Seccareccia F, and Toshima H. The diet and 15-year death rate in the seven countries study. Am J Epidemiol 124: 903–915, 1986.[Abstract/Free Full Text]
  18. Labinskyy N, Csiszar A, Veress G, Stef G, Pacher P, Oroszi G, Wu J, and Ungvari Z. Vascular dysfunction in aging: potential effects of resveratrol, an anti-inflammatory phytoestrogen. Curr Med Chem 13: 989–996, 2006.[CrossRef][ISI][Medline]
  19. Li Y, Schwabe RF, DeVries-Seimon T, Yao PM, Gerbod-Giannone MC, Tall AR, Davis RJ, Flavell R, Brenner DA, and Tabas I. Free cholesterol-loaded macrophages are an abundant source of tumor necrosis factor-alpha and interleukin-6: model of NF-kappaB- and MAP kinase-dependent inflammation in advanced atherosclerosis. J Biol Chem 280: 21763–21772, 2005.[Abstract/Free Full Text]
  20. Maier W, Altwegg LA, Corti R, Gay S, Hersberger M, Maly FE, Sutsch G, Roffi M, Neidhart M, Eberli FR, Tanner FC, Gobbi S, von Eckardstein A, and Luscher TF. Inflammatory markers at the site of ruptured plaque in acute myocardial infarction: locally increased interleukin-6 and serum amyloid A but decreased C-reactive protein. Circulation 111: 1355–1361, 2005.[Abstract/Free Full Text]
  21. Manna SK, Mukhopadhyay A, and Aggarwal BB. Resveratrol suppresses TNF-induced activation of nuclear transcription factors NF-kappa B, activator protein-1, and apoptosis: potential role of reactive oxygen intermediates and lipid peroxidation. J Immunol 164: 6509–6519, 2000.[Abstract/Free Full Text]
  22. Martin-Ventura JL, Blanco-Colio LM, Gomez-Hernandez A, Munoz-Garcia B, Vega M, Serrano J, Ortega L, Hernandez G, Tunon J, and Egido J. Intensive treatment with atorvastatin reduces inflammation in mononuclear cells and human atherosclerotic lesions in one month. Stroke 36: 1796–1800, 2005.[Abstract/Free Full Text]
  23. Marui N, Offermann MK, Swerlick R, Kunsch C, Rosen CA, Ahmad M, Alexander RW, and 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 92: 1866–1874, 1993.[ISI][Medline]
  24. Marui N, Offermann MK, Swerlick R, Kunsch C, Rosen CA, Ahmad M, Alexander RW, and 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 92: 1866–1874, 1993.[ISI][Medline]
  25. Matsumoto T, Komori K, Shoji T, Kuma S, Kume M, Yamaoka T, Mori E, Furuyama T, Yonemitsu Y, and Sugimachi K. Successful and optimized in vivo gene transfer to rabbit carotid artery mediated by electronic pulse. Gene Ther 8: 1174–1179, 2001.[CrossRef][ISI][Medline]
  26. Ohta H, Wada H, Niwa T, Kirii H, Iwamoto N, Fujii H, Saito K, Sekikawa K, and Seishima M. Disruption of tumor necrosis factor-alpha gene diminishes the development of atherosclerosis in ApoE-deficient mice. Atherosclerosis 180: 11–17, 2005.[CrossRef][ISI][Medline]
  27. Pellegatta F, Bertelli AA, Staels B, Duhem C, Fulgenzi A, and Ferrero ME. Different short- and long-term effects of resveratrol on nuclear factor-kappaB phosphorylation and nuclear appearance in human endothelial cells. Am J Clin Nutr 77: 1220–1228, 2003.[Abstract/Free Full Text]
  28. Reiterer G, Toborek M, and Hennig B. Quercetin protects against linoleic acid-induced porcine endothelial cell dysfunction. J Nutr 134: 771–775, 2004.[Abstract/Free Full Text]
  29. Schieffer B, Selle T, Hilfiker A, Hilfiker-Kleiner D, Grote K, Tietge UJ, Trautwein C, Luchtefeld M, Schmittkamp C, Heeneman S, Daemen MJ, and Drexler H. Impact of interleukin-6 on plaque development and morphology in experimental atherosclerosis. Circulation 110: 3493–3500, 2004.[Abstract/Free Full Text]
  30. Spiecker M, Darius H, and Liao JK. A functional role of I kappa B-epsilon in endothelial cell activation. J Immunol 164: 3316–3322, 2000.[Abstract/Free Full Text]
  31. Tzoulaki I, Murray GD, Lee AJ, Rumley A, Lowe GD, and Fowkes FG. C-reactive protein, interleukin-6, and soluble adhesion molecules as predictors of progressive peripheral atherosclerosis in the general population: Edinburgh Artery Study. Circulation 112: 976–983, 2005.[Abstract/Free Full Text]
  32. Ungvari Z, Csiszar A, Edwards JG, Kaminski PM, Wolin MS, Kaley G, and Koller A. Increased superoxide production in coronary arteries in hyperhomocysteinemia: role of tumor necrosis factor-alpha, NAD(P)H oxidase, and inducible nitric oxide synthase. Arterioscler Thromb Vasc Biol 23: 418–424, 2003.[Abstract/Free Full Text]
  33. Ungvari Z, Csiszar A, and Kaley G. Vascular inflammation in aging. Herz 29: 733–740, 2004.[CrossRef][ISI][Medline]
  34. Wang Z, Huang Y, Zou J, Cao K, Xu Y, and Wu JM. Effects of red wine and wine polyphenol resveratrol on platelet aggregation in vivo and in vitro. Int J Mol Med 9: 77–79, 2002.[ISI][Medline]
  35. Wang Z, Zou J, Cao K, Hsieh TC, Huang Y, and Wu JM. Dealcoholized red wine containing known amounts of resveratrol suppresses atherosclerosis in hypercholesterolemic rabbits without affecting plasma lipid levels. Int J Mol Med 16: 533–540, 2005.[ISI][Medline]
  36. Wang Z, Zou J, Huang Y, Cao K, Xu Y, and Wu JM. Effect of resveratrol on platelet aggregation in vivo and in vitro. Chin Med J (Engl) 115: 378–380, 2002.[Medline]
  37. Wood JG, Rogina B, Lavu S, Howitz K, Helfand SL, Tatar M, and Sinclair D. Sirtuin activators mimic caloric restriction and delay ageing in metazoans. Nature 430: 686–689, 2004.[CrossRef][Medline]
  38. Woods A, Brull DJ, Humphries SE, and Montgomery HE. Genetics of inflammation and risk of coronary artery disease: the central role of interleukin-6. Eur Heart J 21: 1574–1583, 2000.[Free Full Text]
  39. Wung BS, Hsu MC, Wu CC, and Hsieh CW. Resveratrol suppresses IL-6-induced ICAM-1 gene expression in endothelial cells: effects on the inhibition of STAT3 phosphorylation. Life Sci 78: 389–397, 2005.[CrossRef][ISI][Medline]
  40. Wung BS, Ni CW, and Wang DL. ICAM-1 induction by TNFalpha and IL-6 is mediated by distinct pathways via Rac in endothelial cells. J Biomed Sci 12: 91–101, 2005.[CrossRef][ISI][Medline]
  41. Yamaoka T, Yonemitsu Y, Komori K, Baba H, Matsumoto T, Onohara T, and Maehara Y. Ex vivo electroporation as a potent new strategy for nonviral gene transfer into autologous vein grafts. Am J Physiol Heart Circ Physiol 289: H1865–H1872, 2005.[Abstract/Free Full Text]
  42. Yeung F, Hoberg JE, Ramsey CS, Keller MD, Jones DR, Frye RA, and Mayo MW. Modulation of NF-kappaB-dependent transcription and cell survival by the SIRT1 deacetylase. EMBO J 23: 2369–2380, 2004.[CrossRef][ISI][Medline]
  43. Zou J, Huang Y, Cao K, Yang G, Yin H, Len J, Hsieh TC, and Wu JM. Effect of resveratrol on intimal hyperplasia after endothelial denudation in an experimental rabbit model. Life Sci 68: 153–163, 2000.[CrossRef][ISI][Medline]
  44. Zou J, Huang Y, Chen Q, Wei E, Cao K, and Wu JM. Effects of resveratrol on oxidative modification of human low density lipoprotein. Chin Med J (Engl) 113: 99–102, 2000.[Medline]



This article has been cited by other articles:


Home page
 Am. J. Physiol. Heart Circ. Physiol.Home page
A. Csiszar, N. Labinskyy, H. Jo, P. Ballabh, and Z. Ungvari
Differential proinflammatory and prooxidant effects of bone morphogenetic protein-4 in coronary and pulmonary arterial endothelial cells
Am J Physiol Heart Circ Physiol, August 1, 2008; 295(2): H569 - H577.
[Abstract] [Full Text] [PDF]

Home page
 Am. J. Physiol. Heart Circ. Physiol.Home page
A. Csiszar, N. Labinskyy, A. Podlutsky, P. M. Kaminski, M. S. Wolin, C. Zhang, P. Mukhopadhyay, P. Pacher, F. Hu, R. de Cabo, et al.
Vasoprotective effects of resveratrol and SIRT1: attenuation of cigarette smoke-induced oxidative stress and proinflammatory phenotypic alterations
Am J Physiol Heart Circ Physiol, June 1, 2008; 294(6): H2721 - H2735.
[Abstract] [Full Text] [PDF]

Home page
 Therapeutic Advances in Respiratory DiseaseHome page
A. Sharafkhaneh, S. Velamuri, V. Badmaev, C. Lan, and N. Hanania
Review: The potential role of natural agents in treatment of airway inflammation
Therapeutic Advances in Respiratory Disease, December 1, 2007; 1(2): 105 - 120.
[Abstract] [PDF]

Home page
 Am. J. Physiol. Heart Circ. Physiol.Home page
M. A. Carluccio, M. A. Ancora, M. Massaro, M. Carluccio, E. Scoditti, A. Distante, C. Storelli, and R. De Caterina
Homocysteine induces VCAM-1 gene expression through NF-{kappa}B and NAD(P)H oxidase activation: protective role of Mediterranean diet polyphenolic antioxidants
Am J Physiol Heart Circ Physiol, October 1, 2007; 293(4): H2344 - H2354.
[Abstract] [Full Text] [PDF]

Home page
 Eur Heart JHome page
L. H. Opie and S. Lecour
The red wine hypothesis: from concepts to protective signalling molecules
Eur. Heart J., July 2, 2007; 28(14): 1683 - 1693.
[Abstract] [Full Text] [PDF]

Home page
 Am. J. Physiol. Heart Circ. Physiol.Home page
Z. Ungvari, Z. Orosz, N. Labinskyy, A. Rivera, Z. Xiangmin, K. Smith, and A. Csiszar
Increased mitochondrial H2O2 production promotes endothelial NF-{kappa}B activation in aged rat arteries
Am J Physiol Heart Circ Physiol, July 1, 2007; 293(1): H37 - H47.
[Abstract] [Full Text] [PDF]

Home page
 Am. J. Physiol. Heart Circ. Physiol.Home page
M. Rajesh, P. Mukhopadhyay, S. Batkai, G. Hasko, L. Liaudet, V. R. Drel, I. G. Obrosova, and P. Pacher
Cannabidiol attenuates high glucose-induced endothelial cell inflammatory response and barrier disruption
Am J Physiol Heart Circ Physiol, July 1, 2007; 293(1): H610 - H619.
[Abstract] [Full Text] [PDF]

Home page
 Am. J. Physiol. Heart Circ. Physiol.Home page
Z. Ungvari, Z. Orosz, A. Rivera, N. Labinskyy, Z. Xiangmin, S. Olson, A. Podlutsky, and A. Csiszar
Resveratrol increases vascular oxidative stress resistance
Am J Physiol Heart Circ Physiol, May 1, 2007; 292(5): H2417 - H2424.
[Abstract] [Full Text] [PDF]

This Article
Right arrow Abstract Freely available
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in ISI Web of Science
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via ISI Web of Science (17)
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Csiszar, A.
Right arrow Articles by Ungvari, Z.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Csiszar, A.
Right arrow Articles by Ungvari, Z.

HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
Visit Other APS Journals Online
Copyright © 2006 by the American Physiological Society.