RT-PCR. Next, a lentiviral shRNA was designed targetingmurine CCR2. This shCCR2 proved to be able to knock-downCCR2 expression at the mRNA and protein levels in vitro.Moreover, migration of SMCs one day after stimulating SMCswith MCP-1 (100 ng/ml), was reduced by 37% in the cellsinfected with lentiviral shCCR2. To demonstrate the efficacy ofthe lentiviral shCCR2 in blocking vein graft disease in vivo, avenous interposition was placed into the carotid artery ofrecipient hypercholesterolemic ApoE3Leiden mice to induceintimal hyperplasia in vivo. In this interposition, intimalhyperplasia with signs of accelerated atherosclerosis developswithin 4 weeks. Directly after graft placement, pluronic gelcontaining 10E8pfu lentiviral shCCR2 or a control lentiviruswas applied around the vein graft. Mice were sacrificed 28 daysafter surgery. Morphometric analysis on sections of graftsshows a significant 38% reduction in total amount of intimalhyperplasia in the shCCR2 treated animals (controls: 0.42±0.05 mm2, shCCR2 :0.26±0.03 mm2, p=0.007). In conclu-sion, these data demonstrate that lentiviral shCC2 reducesmigration of SMCs in vitro. Moreover, local application ofshRNA against CCR2 to the vessel wall prevents intimalhyperplasia and vein graft disease in a hypercholesterolemicmurine in vivo model for vein graft disease. Therefore, this localapplication of RNA interference is very promising astherapeutic tool to prevent vein graft failure.

doi:10.1016/j.vph.2006.08.067

A1.14

Poly (ADP-ribose) polymerase inhibition improvesvascular and contractile function during cardiac rejection

Gabor Szabo, Tamás Radovits, Siegfried Hagl

Dept. of Cardiac Surgery, University of Heidelberg,Heidelberg, Germany

Free radical and in particular peroxynitrite generation andsubsequent activation of poly (ADP-ribose) polymerase (PARP)has been recently shown during cardiac rejection. To determinethe effects of PARP-inhibition during cardiac rejection, isogenicLewis to Lewis (group 1) and allogenic DA to Lewis rat cardiactransplants treated with vehicle (group 2) and with the PARP-inhibitor INO-1001 (group 3) were studied 3 and 5 days afterheterotopic intraabdominal transplantation (n=6/group). Myo-cardial blood flow (MBF) was assessed by the hydrogen-clearance method. Left ventricular systolic pressure (LVSP),maximum pressure development (+dP / dt), end-diastolic pres-sure (LVEDP) were measured by an intraventricular balloon at80 μl volume. Contractile response to dobutamine (5 μg/kg/min) and vasodilatory response to acethylcholine (ACH) andsodium nitroprusside (SNP) were also assessed. Histologicexamination confirmed mild to moderate rejection after 3 daysand severe acute rejection after 5 days in the allogenic groups.

After 3 days, LVSP, +dP / dt and LVEDP showed no significantdifferences between the groups. However, MBF (4.3±0.5 vs.2.2±0.2 vs. 4.1±0.3 ml/min/g, p<0.05) and contractileresponse to dobutamine (+dP / dt: 98±11 vs. 57±7 vs. 88±8%, p<0.05) decreased significantly in group 2 which wasabolished by PARP-inhibition (group 3). While the response toSNP did not differ, the response to ACH was reduced in group 2(78±6 vs. 36±9 vs. 72±7%, p<0.05). After 5 days, LVSP(134±15 vs. 96±11 vs. 115±5 mmHg, p<0.05), +dP / dt(7195±830 vs. 2971±463 vs. 5331±449 mmHg/s, p<0.05)decreased and LVEDP (5.1±0.7 vs. 18.2±3.6 vs. 8.3±1.0 mmHg, p<0.05) increased in group 2 which was partlyreduced by PARP inhibition (group 3). While MBF, response todobutamine and ACH did not change in group 1, they decreasedin groups 2 and 3 (p<0.05). Thus, contractile and vasomotordysfunction occur in a typical time dependent manner duringcardiac rejection which can be reduced by PARP-inhibition.PARP-inhibition may represent a novel therapeutic approach toreduce functional disturbances during cardiac rejection.

doi:10.1016/j.vph.2006.08.068

A1.15

Prognostic value of plasma N-terminal pro-brainnatriuretic peptide, interleukin 6, tumor necrosis factoralpha and soluble CD40 ligand in patients acute heartfailure

Radek Pudil1, Milos Tichy2, Ctirad Andrys3, MarcelaDrahosova3, Vaclav Blaha4, Jan Vojacek1

1Charles University Prague, Medical Faculty HradecKralove, Hradec Kralove, Czech Republic2Institute of Clinical Biochemistry and Diagnostics, CharlesUniv, Hradec Kralove, Czech Republic3Institute of Clinical Immunology and Allergology, Med Fac,Hradec Kralove, Czech Republic4Dept of Medicine, Faculty of Health Sciences, HradecKralove, Czech Republic

Plasma natriuretic peptide levels and cytokine (interleukin6, IL6, tumor necrosis factor alpha, TNFa) levels are increasedin heart failure. However, little is known about NT-proBNP,cytokine and sCD40L levels and their correlation with themortality. We assessed the relationship between plasma IL 6,TNFa, sCD40L and N-terminal pro-brain natriuretic peptide(NT-proBNP) levels and short and long term mortality from allcauses in patients with acute heart failure.NT-proBNP, IL 6,TNFa and soluble CD40 ligand were measured in baselinesamples from 92 consecutive patients referred for acute heartfailure to intensive care department. During one-year follow-up, 32 patients died. Mean plasma levels of NT-proBNP, IL 6and TNFa were significantly lower among the patients whosurvived (NT-proBNP: 7 855.4±9919.9, vs.15470.6±11273.1,

e714th IVBM Abstracts