Dietary enrichment of apolipoprotein E-deficient mice with extra virgin olive oil in combination with seal oil inhibits atherogenesis.

BACKGROUND: In this study we investigated the antiatherogenic effect of dietary enrichment of a combination of extra virgin olive oil (EVO) and seal oil on apolipoprotein E-deficient (apoE-/-). METHODS: Six-week-old female and male apoE-/- mice were for 12 weeks fed a lipid rich diet containing 19.5% fat and 1.25% cholesterol without any supplement, with 1% (wt/wt) mixture of extra virgin olive and seal oil (EVO/n-3), or 1% corn oil, respectively. RESULTS: Supplementation with the combination of EVO/n-3 significantly reduced atherosclerotic lesion formation in the aortic arch, thoracoabdominal, and total aorta of female apoE-/-mice. The effect was less pronounced in male mice and significant reduction was only observed in the thoracoabdominal region of the aorta. There were no differences or changes in dietary intake or body weight gain. However, compared to the other groups, plasma levels of triglycerides were reduced in both female and male mice fed the EVO/n-3 mixture. Male mice on both treatments showed reduced plasma cholesterol compared to the control mice after 12 weeks on diet. CONCLUSION: Dietary supplementation of a marine/olive oil combination inhibits atherosclerotic lesion formation in the female apoE-/- mice by antithrombotic, antihypertriglyceridemic, and antioxidant effects.

Seafood diets: hypolipidemic and antiatherogenic effects of taurine and n-3 fatty acids.

BACKGROUND: Health aspects of seafood have primarily been linked to n-3 polyunsaturated fatty acids (PUFA), eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA). Although animal studies have suggested beneficial contributions from taurine, highly abundant in seafood, its effect in humans is obscure. This study evaluates the combined effects of n-3 PUFA and taurine. METHODS: Healthy volunteers (n=80) were recruited to a 7-week double-blind and parallel intervention trial. One group (n=39) received fish pâté (36g/day) enriched in n-3 (1.1gEPA+DHA/day) and the second (n=41) an identical pâté enriched both in n-3 and taurine (425mg/day). RESULTS: Total cholesterol (TC) (-5%, P<0.001), low-density lipoprotein (LDL)-cholesterol (-8%, P<0.001) and Apo B (-4%, P<0.001) decreased more in the n-3+taurine compared to the n-3 group. A significant within-group enhancement of high-density lipoprotein (HDL)-cholesterol was demonstrated in the n-3+taurine group (6%, P<0.0001). Reductions in triacylglycerol (TG) (-16%, P<0.05 in n-3; -14%, P<0.05 in n-3+taurine), thromboxane B(2) (TxB(2)) (-21%, P<0.001 in n-3; -15%, P<0.05 in n-3+taurine), tumor necrosis factor (TNFalpha) (-24%, P<0.001 in n-3; -12%, P<0.05 in n-3+taurine) and monocyte chemotactic protein (MCP-1) (-12%, P<0.05 in n-3; -6%, P<0.0001 in n-3+taurine) were evident in both groups. Reductions in interleukin (IL)-6 (-16%, P<0.05) and LTB(4) (-18%, P<0.05) were only significant in the n-3 group. CONCLUSIONS: The effects, particularly on blood lipids, of combining n-3 PUFA's and taurine proved superior to those of n-3 alone.

Granulocytes do not express but acquire monocyte-derived tissue factor in whole blood: evidence for a direct transfer.

Unlike unanimous opinion on tissue factor (TF) expression in monocytes, the quest for TF presence in granulocytes has been going on for decades. To study the cell origin and track the blood-borne TF, we assessed TF activity and protein levels, knocked-down endogenous TF expression with small interfering RNA (siRNA), and overexpressed TF-yellow fluorescent protein (TF-YFP) fusion in immunologically isolated human monocytes and granulocytes. Monocytes and, to a much lesser extent, granulocytes isolated from lipopolysaccharide (LPS)/phorbol 12-myristate-13-acetate (PMA)-stimulated whole blood contained active TF antigen. However, only monocytes possessed significant TF activity and protein levels when stimulated with LPS/PMA in suspension. Reintroduction of TF-silenced monocytes to whole blood led to a profound reduction of LPS/PMA-stimulated TF activity in both mononuclear cell (MNC) and granulocyte fractions. No reduction in TF activity in MNC and granulocyte fractions was observed when TF-silenced granulocytes were reintroduced to whole blood. As shown by immunoblotting, flow cytometry, and confocal microscopy, granulocytes became positive for TF-YFP when isolated from whole blood reconstituted with TF-YFP-expressing monocytes. Together, we pinpoint monocytes as a major source of TF and provide solid experimental evidence for a direct transfer of TF protein from the monocytes to granulocytes in the blood.

Induction of monocytic tissue factor expression after rewarming from hypothermia in vivo is counteracted by heat shock in c-Jun-dependent manner.

OBJECTIVE: Triggering of tissue factor (TF)-mediated blood coagulation leads to the development of disseminated intravascular coagulation during rewarming from hypothermia. We studied post-rewarming TF levels, activity, and surface redistribution, along with the regulation of TF gene transcription in mononuclear cells (MNCs) obtained from an in vivo rat model. METHODS AND RESULTS: Rewarming after a 5-hour episode of 15 degrees C hypothermia caused an increase in TF activity, protein levels, and externalization of TF antigen in rat MNCs. This was accompanied by a dramatic elevation of c-Jun and JNK phosphorylation, and the absence of EGR-1 and NF-kappaB activation. To search for a stimulus to counteract c-Jun-mediated induction of TF activity in MNCs from rewarmed rats, we applied heat shock pretreatment one day before the hypothermia/rewarming experiment. This restored post-rewarming TF activity, protein levels, and surface-to-total TF ratio in rat MNCs to normothermic levels. Furthermore, in heat shock-pretreated animals, rewarming failed to increase phosphorylated c-Jun and JNK levels. We attribute this to the profound overexpression of heat shock protein 70 and inhibition of JNK. CONCLUSIONS: MNCs respond to rewarming from hypothermia by an induction of active TF antigen. This effect is dependent on c-Jun activation and is abolished by heat shock pretreatment.

Intracellular and surface distribution of monocyte tissue factor: application to intersubject variability.

OBJECTIVE: The high and low responder phenomenon describes individual differences in lipopolysaccharide (LPS)-induced monocyte tissue factor (TF) activity. We characterized patterns of intracellular accumulation, externalization, and shedding of TF in response to LPS in mononuclear cells (MNCs) from high responders (HRs) and low responders (LRs). METHODS AND RESULTS: After 2 hours of LPS stimulation of whole blood, flow cytometry analyses revealed a larger population of TF-positive monocytes in HRs (32.0+/-3.5%) versus LRs (11.2+/-1.2%; P< or =0.05), along with a stronger mean fluorescence intensity of TF signal in HRs (7.1+/-0.5 arbitrary units [AU]) compared with LRs (5.4+/-0.4 AU; P< or =0.05). The LPS-treated blood of the HR group contained 2-fold more TF-positive microparticles than LRs. In-cell Western assay demonstrated higher intracellular accumulation of TF in mononuclear cells (MNCs) from LRs because LPS induced a 3.7-fold increase of total TF levels in LRs versus a 1.5-fold increase in HRs. In contrast, in response to LPS stimulation, MNCs from HRs exhibited a 4-fold induction of surface TF, whereas MNCs from LRs only had a minor increase in surface TF levels. CONCLUSIONS: The higher availability of surface TF antigen on MNCs from HRs and TF-containing microparticles might make these individuals more susceptible to hypercoagulation.