ceptor B1 (SR-B1), caveolins ATP-binding membrane cassette trans-porter A-1 (ABCA-1), and sterol 27-hydroxylase (CYO27A1) werealso measured by real-time PCR and western blot. Results: CRPsignificantly reduced cholesterol efflux to apoA1 from humanmacrophage-derived foam cells in a concentration-dependent man-ner. A clinical relevant concentration of CRP (10 ng/ml) scientificallyreduced cholesterol efflux by 27% as compared to controls (P�0.05).An increased accumulation of intracellular cholesterol in the CRP-treated foam cells was observed by Oil red staining. Furthermore,CRP (10 ng/ml) significantly reduced mRNA levels of ABCA-1 by 35%in macrophage-derived foam cells as compared to controls (p�0.05).In parallel, protein levels of ABCA-1 were also significantly reducedin the CRP-treated cells (P�0.05). However, there were no signifi-cant changes for SR-B-1, caveolins, and CYO27A1 between thetreated and control groups. Conclusions: These data demonstrate,for the first time, that CRP significantly inhibits cholesterol effluxfrom human macrophage-derived foam cells in vitro. Down-regulation of intracellular cholesterol transport molecule ABCA-1may be a possible mechanism of CRP-induced inhibition of choles-terol efflux, which may contribute to inflammation- associated ath-erosclerosis.

116. EFFICACY OF PERIVASCULAR NO-ELUTING THERA-PIES FOR THE PREVENTION OF NEOINTIMAL HY-PERPLASIA. Najjar SF, Pearce C, Eng J, Aalami O,Kapadia M, Lyle B, Murar J, Jiang Q, Kibbe MR; NorthwesternUniversity

INTRODUCTION: Nitric oxide (NO) has been shown to effec-tively inhibit neointimal hyperplasia in animal models of arterialinjury and vein bypass grafting. However, none of these previoustherapies have been deployed in the clinical arena, mostly due tosafety concerns, side effects, complexity or lack of significant anddurable effect. Recent advances in NO donor chemistry have pro-vided donors with more stable and longer eluting profiles. The aim ofthis study is to examine the efficacy of two perivascular diazenium-diolate NO-eluting therapies (PROLIN/NO and PAN/NO) that haveclinical potential at inhibiting balloon injury-induced neointimal hy-perplasia in rat carotid arteries. PROLI/NO releases a large amountof NO over a short duration, while PAN/NO releases a lower steadystate of NO over a prolonged duration. METHODS: The standardrat carotid artery injury model was utilized. Following arterial injurywith the 2F embolectomy catheter, PROLI/NO, PAN/NO or PROLI/NO�PAN/NO, were applied to the external surface of the artery,either in powder form (20mg) or dissolved in a polylactic-polyglycolicacid polymeric matrix (Polaxamer) at 16.7% (n�4-5 per group). Thispolymer is liquid at 4 degrees and gel at 37 degrees. Animals weresacrificed at 2 weeks and the arteries were perfusion fixed in situ.Morphometric analysis was performed on routine hematoxylin andeosin stained sections. Intimal (I) and medial (M) area were calcu-lated on five equally spaced sections throughout the injured segmentand the I/M area ratio was calculated. The degree of inflammationwas assessed using immunohistochemical staining for neutrophils,monocytes, and lymphocytes with antibodies against PMN, ED1, andCD45, respectively. The effect of the treatment groups on cellularproliferation was assessed immunohistochemically with an antibodyagainst Ki67, a marker for actively proliferating cells. For bothinflammation and proliferation, the slides were scored on a scalefrom 0-3 by five independent blinded observers and the average scorewas reported. All data is expressed as the mean �/- the standarderror of the mean. Statistical significance was assumed for P�0.05.RESULTS: The Injury alone and Poloxamer alone groups were usedas the controls for the powder and gel treatment groups, respectively.Treatment with Poloxamer alone resulted in a slight increase in theI/M area ratio (1.08�0.06) compared to injury alone (I/M 0.964�0.04,P�NS). Treatment with PAN/NO gel and powder reduced the I/Marea ratio by 46% and 67% compared to their controls (I/M0.589�0.07 and 0.323�0.05, respectively, P�0.05). Treatment with

PROLI/NO gel reduced the I/M area ratio by 69% (0.33�0.06,P�0.05). Most dramatically, treatment with PROLI/NO powder re-duced the I/M area ratio by 86% (0.138�0.05, P�0.05). Of note, thisrepresents a 91% reduction in the intimal area alone (12,657PROLI/NO vs 142,496 injury, P�0.05). The combination therapy ofPROLI/NO � PAN/NO powder resulted in an I/M area ratio reduc-tion of 80% compared to injury alone (0.195�0.07, P�0.05). Alltreatment groups that received PAN/NO stained strongly positive formarkers of inflammation (neutrophils, monocytes, lymphocytes).Conversely, minimal to no staining for markers of inflammation wasobserved in the PROLI/NO treatment groups. PAN/NO andPROLI/NO both inhibited VSMC proliferation in the neointima andmedia equally. CONCLUSIONS: Perivascular delivery of diazeni-umdiolate NO-donors in a powder or polymer-based gel is an effec-tive method of preventing the development of neointimal hyperplasiafollowing arterial injury. Of the therapies examined, PROLI/NO wasthe most effective at inhibiting neointimal hyperplasia and inflam-mation. Overall, this NO-eluting therapy is simple to use and rep-resents a potentially clinically applicable therapy to prevent thedevelopment of neointimal hyperplasia following arterial injury.

117. C-REACTIVE PROTEIN INCREASES PLASMINOGENACTIVATOR INHIBITOR-1 EXPRESSION OF HUMANENDOTHELIAL CELLS. Chen CJ, Nan B, LinPH,Lumsden AB, Yao QC; Baylor College of Medicine

Background: C- reactive protein (CRP) has been shown to be aninflammatory marker which predicts cardiovascular disease. How-ever, little is known whether CRP has direct effects on endothelialfunctions and gene expression. The purpose of this study was todetermine the effects and molecular mechanisms of CRP on theexpression of plasminogen activator inhibitor-1 (PAI-1), an impor-tant mediator of vascular disease, in human coronary artery endo-thelial cells (HCAECs). Methods: HCAECs were treated with CRPin a time- and dose-dependent manner. The mRNA levels of PAI-1and CD32, a CRP receptor, were determined by real-time PCR. PAI-1protein and activities were investigated by ELISA, western blottingand specific chemical reaction. Activation of mitogen-activated pro-tein kinases (MAPKs) was assessed with Bio-Plex immunoassay.Anti-CD32 antibody and dietary antioxidant curcumin were used toblock the action of CRP. The effect of CRP on PAI-1 expression wasalso tested in other types of human endothelial cells. Results:HCAECs treated with CRP at a range of clinically relevant concen-trations (5, 10 and 25 �g/ml) for 12 hours significantly increasedPAI-1 mRNA levels by 24, 49 and 83%, respectively, as comparedwith controls (P � 0.05). The maximal effect of CRP (10 �g/ml) wasobserved at 12 hours. The protein level and enzyme activity of PAI-1in the supernatant of CRP (10 �g/ml)-treated HCAEC cultures weresignificantly increased by 144% and 64%, respectively, at 12 hours ascompared with controls (P�0.05). Cell-associated PAI-1 protein lev-els were also significantly increased. In addition, CRP (10 �g/ml)significantly increased CD32 mRNA levels by 277% as comparedwith controls (P � 0.05). CRP substantially increased phosphoryla-tion of MAPK p38, but not JNK and ERK1/2. Furthermore, anti-CD32 antibody blocked CRP-induced PAI-1 mRNA expression from148% to 110% (P�0.05). Curcumin (5 �M and 10 �M) dramaticallyreduced PAI-1 mRNA levels by 50 and 42%, respectively, as com-pared with controls (P � 0.001). The effect of CRP on PAI-1 expres-sion was also confirmed in other types of human endothelial cellsisolated from umbilical vein, skin, and lung microvessels. Conclu-sions: CRP significantly increased the expression of PAI-1 in a time-and dose-dependent manner in HCAECs and other human endothe-lial cells. CRP also increased its receptor CD32 expression whichmay further enhance the action of CRP. CRP-induced PAI-1 expres-sion may be mediated by oxidative stress and p38 signal pathway asantioxidant curcumin effectively blocks the effect of CRP onHCAECs.

205ASSOCIATION FOR ACADEMIC SURGERY AND SOCIETY OF UNIVERSITY SURGEONS—ABSTRACTS