Tu-P7:210 Hypoxia induces vasucular inflammatory by upregulate OxLDL receptor

Tues&ty, June 20, 2006: Poster Session P7 Basic science ( lst part) Methods: Primary culture of human macrophages was used for evaluation of anti-atherogenic effects of inflaminat. The ability of serum that was obtained before single dose oral administration of drugs and 2, 4 and 8 hours after it to induce cholesterol accumulation (serum atherogenidty) was measured. Didofenac was used for comparison. Results: At the first stage of this study we tested anti-cytokine effect of inflaminat. Administration of inflaminat led to the substantial decrease of serum ability to induce over-expression of inflammatory markers IL-1 and TNF-a in cultured cells, quite comparable with effect of diclofenac. Then antiatherogenic properties of inflaminat were investigated. Human serum induced 1.3-1.8-fold increase of cholesterol level in cultured cells. After administration of inflaminat, serum atherogenicity was lowered by 61.84-7.3% after 2 hours, by 71.94-7.0% at 4 hours, by 58.04-9.1% at 8 hours. Diclofenac administration did not resulted in significant changes in serum atherogenicity. Conclusions: Natural anti-inflammatory drug inflaminat provides anticytokine effect. At the same time, it suppresses intracellular cholesterol deposition. Double mechanism of inflaminat action allows to develop novel clinical implication for atherosclerosis prevention and treatment. I

I Tu-P7:210 Ii H Y P O X I A I N D U C E S V A S U C U L A R INFLAMATION BY U P R E G U L A T E OXLDL R E C E P T O R S. Ogura;, T. Shimosawa 2, H. Matsui;, T. Sawamura;, T. Fujita;.

1Department of Nephrology arm EtMocrinology, Universi~ of Tokyo, Tokyo, Japan: 2Faculi~ of Medicine, Univesi~ of Tokyo, Tokyo, Japan Objective: Chronic hypoxia is one of the major causes of vascular remodeling associated with stimulating reactive oxygen species (ROS) production. An excess of ROS has been suggested to trigger for atherosclerosis, and ROS upregulates expression of several factors to modify vascular remodeling. Lectin-like oxidated low density lipoprotein receptor -I(LOX-1) is a indentiffed endotherial receptor for oxidized low-density lipoprotein (oxLDL) and assumed to play a role in the initiation and progression of atherosclerosis. In previous study, LOX-1 gene was regulated by oxidative stress. In this study, we studied LOX-1 receptor expression could be change in hypoxia in mouse lung and microvascular endothelial cells. Metbod and Results: Immnoblot assay and RT-PCR demonstrated that LOX-1 receptor was increased in hypoxia. OxLDL uptake were also inceased in hypoxic cultures. Hypoxia increased production of ROS production in murine lung tissue, measured by lucigenin chemiluminescense and immunostaining of 3-nitrotyrosine. Hypoxia enhanced LOX-1 receptor mRNA expression, which was reduced by the administration of a superoxide dismutase mimetic (hydroxy-TEMPO) and NADPH oxidase inhibitor (apocynin). Conclusion: Hypoxia increased LOX-1 receptor expression, which is sensitive for reactive oxygen spieces, suggesting a potential role in atherosclerosis.

I Tu-P7:211 I M O R E I N F L A M M A T I O N IN FAMILIAL H Y P E R C H O L E S T E R O L E M I A T H A N IN FAMILIAL C O M B I N E D HYPERLIPIDEMIA P. Pauciullo, M. Gentile, G. Marotta, S. Ubaldi, F. Jossa, G. Iannuzzo, F. Faccenda, A. Baiano, C. Mormile, E Rubba. Dept. of Cl#dcal and

Eaperimental Medicbw, Universi~ FedeHco II, Naples, Italy Objective: To test the role of low-density lipoprotein cholesterol (LDL-C) as an independent contributor to low-grade inflammation as expressed by an high-sensitive C-reactive protein (HS-CRP) level > 3 mg/1 and < 15 mg/1 in familial dyslipidemias. Patients and Methods: The relationship between LDL-C and HS-CRP was measured by logistic analysis in a sample of 120 patients with familial hypercholesterolemia (FH) (44% men), 137 patients with familial combined hyperlipidemia (FCHL) (44% men), and 146 unrelated control subjects (42% men) recruited in the Lipid Clinic of the University Federico II in Naples. Results: LDL-C > 160 mg/dl was associated with an increased likelihood of a HS-CRP > 3 mg/1 (P = 0.010). This relationship holds after adjustment for potential confounders such as sex, age tertiles, high-density lipoprotein cholesterol, body mass index, and homeostasis model assessment. The relationship between LDL-C and HS-CRP explains why in young (< 40 years of age) familial dyslipidemic patients higher values of HS-PCR were found in FH patients (mean LDL-C 289 mg/dl) as compared with FCHL patients (mean LDL-C 208 mg/dl) (P=0.023). Conclusion: Present data indicate that LDL-C plays an inflammatory role at least as relevant as that played by the markers of insulin resistance.

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Tu-P7:212 ] ASPIRIN A N D S A L I C Y L A T E SUPPRESS P O L Y M O R P H O N U C L E A R APOPTOSIS DELAY M E D I A T E D BY P R O I N F L A M M A T O R Y S T I M U L I S. Ne,orotto;, E. Malaver;, N. Pacienza;, L.P. D'Atri;, P. Urdinez;, R.G. Pozner;, R.M. Gomez 2, M. Schattner;. 1National Academy of

Medicine, Buenos Aires, Argentina: Argentitta

2 Universi~

of la Plata, la Plata,

Objectives: Aspirin (ASA) and sodium salicylate (NaS) are widely drugs used to treat inflammation. During inflammation, several factors prolong polymorphonuclear leukocytes (PMN) survival. We examined whether salicylates interfere with PMN apoptosis. Methods: Apoptosis was assessed by cell staining with acridine orange and ethidium bromide (fluorescence microscopy) or with propidium iodide or annexin-V (flow cytometry). IkB alpha degradation was measured by western blot. Phagocytosis was quantified by May Grunwald-Giemsa. Results: PMN preincubation with ASA or NaS suppressed LPS (0.5ug/ml) (EC50: 1.35mM and 1.53mM) or IL-1 alpha (1.39mM and 1.28mM, n=6) prolonged PMN survival through inhibition of IkB alpha degradation. Although ASA and NaS did not trigger PMN death (1-3raM, n=5), both drugs significantly increased Zymosan-mediated apoptosis (15, 22, 28% of control, 1 and 3raM ASA or NaS, n=4). Indomethadn and ibuprofen did not inhibit LPS antiapoptotic activity (n=4). The cytokines-prolonged PMN survival was no longer observed in PMN from donors who had ingested ASA (2g, n=7). Using a thioglycolate-induced peritonitis model, we observed that in ASA or NaS-treated mice there was not only a decrease in the number of total cells recruited (1.34-0.3, 1.74-0.4 x10(6) cells/ml) but also an increase in both, percentage of apoptotic PMN (284-5, 234-2%) and enhanced macrophages phagocytosis (454-4, 484-6%) compared with controls (3.34-0.3, 94-2, 174-5 respectively, n=5). Conclusion: Another mechanism by which salicylates exert its antiinflammatory action is by shortening PMN life span. Funding: National Agency for Sdentific Promotion.

Tu-P7:213 ] R E L A T I O N S H I P S B E T W E E N A U T O A N T I B O D I E S AGAINST MODIFIED LDL A N D ADVANCED GLYCATION END-PRODUCTS IN CLINICALLY HEALTHY M E N E Slogren , G.N. Fredrlkson-, G. Basta 3, A. Hamsten I , J. Nllsson-, M.L. Hellenius 4 , R. De Caterina 3 , R.M. Fisher 1. 1King Gustaf VResearch

bzstitute, Karolinska bzstitutet, Stockhohn, Sweden: 2Department of Clinical Sciences, Malta5 Universi~ Hospital, MalmS, Sweden: 3CNR btstitute of Clinical Physiology,~ Pisa, Italy: 4Center for Family attd Communi~ Medicbw, Karolbtskxt Instituter, Stockhohn, Sweden Objective: Advanced glycation end products (AGEs) have been proposed to contribute to increased risk of atherosclerotic disease in insulin resistant individuals, partly by prompting inflammatory responses. Autoantibodies against modified LDL might play important roles in the genesis of atherosclerosis, but clinical relationships between plasma AGEs, inflammatory markers and autoantibodies against modified LDL are presently unclear. Methods: 291 healthy men (63 y) with a range of insulin sensitivities were investigated. Circulating IgG and IgM antibodies against aldehyde modified peptide sequences, denoted 37, 45, 210 and 245, within a p t B-100 were quantified. Circulating oxidised LDL, inflammatory markers and carboxy-methyl lysine (CML) (the main AGE in human plasma) were quantified by ELISA. Fasting insulin, lipid and lipoprotein concentrations were also determined. Results: Plasma CML was positively related to IgG and IgM autoantibodies agadnst two specific epitopes of a p t B-100; IgG210, r=0.16, p<0.01; IGG245, r=0.20, p<0.01; IgM210, r=0.13, p<0.05 and IGM245, r=0.13, p<0.05. Plasma CML was related to tumor necrosis factor alpha (r=0.14, p<0.05), but not to C-reactive protein or interleukin-6. Furthermore, plasma CML was related to HDL cholesterol (r--0.20, p<0.01) and the ratio oxidised LDL/LDL cholesterol (r=0.14, p<0.05), but not to fasting plasma insulin or glucose. Condusions: These findings suggest that increased levels of plasma AGEs might promote the modification of LDL and thereby trigger the production of specific autoantibodies agadnst certain epitopes of a p t B-100. Funding: No commercial interests

XIV bztetTtational Symposium on Atherosclerosis, Rome, Italy, June 18-22, 2006