Eicosapentaenoic acid ameriolates postprandial hypertriglyceridemia

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Abstracts / Atherosclerosis 235 (2014) e84–e191

Methods: Serum levels of non-cholesterol sterols (phytosterols, cholesterol precursors and bile acids precursors) were measured by HPLC-MS/MS before and after lipid-lowering treatment in a 43 years-old woman, and in 10 normolipemic age-matched women. Results: The patient reported jaundice, fatigue and generalized pruritus in the last 6 months. She had no personal or family history of hyperlipidemia or premature cardiovascular disease. Previous total cholesterol was in the normal range. Physical examination revealed jaundice, interdigital xanthomas, xanthelasmas and palmar xanthomas. Sonography of Achilles tendons did not show enlargement or lipid accumulation compatible with tendon xanthomas. Her total cholesterol was 1404 mg/dL; triglycerides 146 mg/dL; HDL cholesterol 58 mg/dL; conjugated bilirubin 2.3 mg/dL; alkaline phosphatase 1362 U/L. APOE genotype: E3/E3. The diagnosis of PBC was confirmed with the presence of antimitochondrial antibodies, and compatible histological liver findings. Two months after treatment with statins, fenofibrate and ezetimibe her total cholesterol level was 516 mg/ dL; triglycerides 159 mg/dL; HDL cholesterol 96 mg/dL; conjugated bilirubin 2.07 mg/dL; alkaline phosphatase 1089 U/L. Non-cholesterol sterol were highly elevated especially for cholestanol and plant sterols that were 10-30 times higher in the patient than in controls, even after ezetimibe treatment. Cholesterol precursors (desmosterol and lanosterol) were in the normal range after adjustment for total cholesterol and decreased, as expected, with statin treatment. Desmosterol/cholestanol and desmosterol/sitosterol ratios, good surrogates of hepatic synthesis/intestinal absorption balance, were highly decreased in the patient. Conclusion: The severe hypercholesterolemia and xanthomatosis of this patient are extremely rare. This case emphasizes the critical role of biliary excretion in the cholesterol, cholestanol and phytosterols metabolism; the profound effect of PBC in their biliary secretion, suggesting a dysfunctional effect on ABCG5/ABCG8 transporters, and arises some uncertainness about biliary cholesterol excretion in humans. 07 - Role of the intestine for lipid homeostasis and atherosclerosis EAS-0612. TTC39B DEFICIENCY PROMOTES HDL PRODUCTION AND IMPAIRS NONHDL ABSORPTION IN SMALL INTESTINE M. Kosekia, J. Hsiehb, E. Yakushijib, C. Welchb, J. Iqbalc, M.M. Hussainc, S. Takiguchid, D.J. Raderd, Y. Sakataa, S. Yamashitaa, A.R. Tallb a Cardiovascular Medicine, Osaka University Graduate School of Medicine, Osaka, Japan; b Molecular Medicine, Columbia University Medical Center, New York, USA; c Cell Biology, State University of New York Health Science Center at Brooklyn, New York, USA; d Translational Medicine, University of Pennsylvania School of Medicine, Philadelphia, USA

Objectives: Recently, human genome wide association studies (GWASs) have led to the discovery of novel genes and loci associated with alterations in plasma triglyceride (TG), low-density lipoprotein (LDL-C) or highdensity lipoprotein cholesterol (HDL-C) levels. TTC39B (T39) was identified in a GWAS as a novel gene influencing HDL cholesterol (HDL-C) levels. Methods: We have evaluated lipid profile and cholesterol absorption in T39-/- mice on chow or high cholesterol/fat/bile salt diet. We also examined the susceptibility of steato-hepatitis in long term study with the high cholesterol/fat/bile salt diet. Furthermore, we investigated atherosclerosis in T39-/-, Ldlr-/- mice fed high cholesterol diet. Results: On a chow diet, HDL-C levels were significantly increased by 22%. There were increases in LXR protein but not mRNA, increased expression of ABCA1 mRNA and protein and increased secretion of HDL in small intestinal enterocytes. When mice were challenged with a high fat/high cholesterol/bile salt diet, there was a significant 42% increase in HDL-C and also decreased incorporation of dietary cholesterol and fat into chylomicrons and marked protection from steato-hepatitis. In addition to intestinal changes, there was increased LXR protein and induction of Abcg5/8 in liver. T39-/-, Ldlr-/- mice on high cholesterol diet showed increased HDL-C, decreased VLDL/LDL-C and decreased atherosclerosis. Conclusion: These studies show that T39 deficiency results in increased LXR primarily in enterocytes, beneficial lipoprotein changes and reduced atherosclerosis.

07 - Role of the intestine for lipid homeostasis and atherosclerosis EAS-0636. DIOSGENIN INDUCES TRANS-INTESTINAL CHOLESTEROL EXCRETION IN LDL RECEPTOR DEFICIENT FEMALES J.M. Bergera, F. Moreaua, X. Prieura, P. Costeta, F. Lasserreb, H. Guilloub, B. Carioua, C. Le Maya a b

Institut du Thorax, INSERM UMR 1087/ CNRS UMR 6291, NANTES, France; TOXALIM INRA, UMR 1331 TOXALIM INRA, Toulouse, France

Objectives: Aim. Transintestinal cholesterol excretion (TICE) is an alternative pathway to biliary cholesterol excretion and accounts for 33% of fecal cholesterol excretion in mice. TICE is inducible by several pharmacological drugs or nutrients. The plant sterol diosgenin exerts hypocholesterolemic properties at least by promoting biliary cholesterol excretion and reducing intestinal cholesterol absorption. We aimed at characterizing the effect of diosgenin on TICE. Methods: C57Bl/6J (WT) and low-density lipoprotein receptor knock-out (LDLR-KO) female mice received a standard chow diet enriched or not with 1% diosgenin (wt/wt) for 10 days. TICE was measured in vivo in mice injected i.v. with radiolabelled LDL, by cannulation of the proximal intestine and concomitant surgical bile diversion, counting radioactivity in intestinal perfusates over 120 minutes. Results: Plasma total cholesterol was significantly reduced in diosgeninfed WT and LDLR KO mice respectively from 0.83
0.05 to 0.67
0.05 and from 1.48
0.06 to 1.17
0.05 g/l. Interestingly, FPLC analysis revealed that diosgenin specifically decreased circulating LDL cholesterol in LDLRKO mice. In both genotypes, fecal cholesterol excretion was strongly induced by diosgenin. We confirmed that intestinal cholesterol absorption was lowered by diosgenin in WT (-55,2%, P<0,05) and LDLR-KO (-37,7%, P<0,05) mice. In WT mice, neither biliary cholesterol excretion nor TICE were significantly induced in response to diosgenin-enriched diet despite a trend for increase (respectively + 79,8% and + 34,3%). In contrast, both of these pathways were significantly stimulated by diosgenin in LDLR-KO mice (respectively +165% and +55,1%). Gene expression analyses indicated that diosgenin increased the hepatic and intestinal expression of several Pregnane X Receptor (PXR) target genes. However, the hypocholesterolemic effect of diosgenin was maintained in PXR-KO mice. Conclusion: We show that diosgenin is a new TICE modulator. TICE activation contributes to the hypocholesterolemic effect of diosgenin in LDLRKO mice. 07 - Role of the intestine for lipid homeostasis and atherosclerosis EAS-0174. EICOSAPENTAENOIC ACID HYPERTRIGLYCERIDEMIA

AMERIOLATES

POSTPRANDIAL

D. Masudaa, T. Kobayashia, T. Okadaa, H. Nakaokaa, R. Kawasea, K. Nakatania, M. Kosekia, T. Ohamab, M. Nishidab, Y. Sakataa, S. Yamashitac a Department of Cardiovascular Medicine, Osaka University Graduate School of Medicine, Suita Osaka, Japan; b Health Care Center, Osaka University, Toyonaka Osaka, Japan; c Department of Community Medicine, Osaka University Graduate School of Medicine, Suita Osaka, Japan

Objectives: Postprandial hypertriglyceridemia (PHTG) is supposed to be caused by the postprandial accumulation of chylomicrons (CM) produced from intestine and their remnants. Since CM remnants are highly atherogenic, interventional therapy has long been investigated for improving PHTG using anti-hyperlipidemic drugs. Recently, one of polyunsaturated fatty acids, eicosapentaenoic acids (EPA), have been used for treatment of hypertriglyceridemia, however the effect of EPA on PHTG and its mechanism have not been clear. In the current study, we investigated the effect of EPA on PHTG focusing on intestinal CM production.

Abstracts / Atherosclerosis 235 (2014) e84–e191

Methods: Male C57BL/6J mice (8 weeks old) were fed a western diet (WD) for 2 weeks, and 3 groups of mice (n¼10/group) were fed a WD along with forced oral dosage of saline, olive oil or EPA (0.2 g/100 g body weight/day) for another 2 weeks. Oral fat loading (OFL) test was performed using olive oil by gavage (17 mL/g body weight) after an overnight fast. The changes in plasma levels of total cholesterol (TC), TG, free fatty acids (FFA) and apoB48 mass were compared during the fasting state and at 2, 4, and 6 hours after OFL. Postprandial lipoprotein profiles of plasma were analyzed by high performance liquid chromatography (HPLC). Results: There was no significant difference in both dietary intake and weight gain among the 3 groups. Concentrations of TG, FFA and TC and apoB-48 mass after OLF were significantly lower in EPA group than in saline or olive oil groups (the incremental area under the curve of TG, saline vs olive oil vs EPA, 1312.9
446.4 vs 1434.9
308.1 vs 867.7
173.4 mg*6h/dl; p<0.01). HPLC analysis showed that peaks of lipoproteins in the size range of CM and VLDL were decreased in EPA group compared with the saline and olive oil groups. Conclusion: EPA may attenuate PHTG by reducing intestinal TG and chylomicron production. 07 - Role of the intestine for lipid homeostasis and atherosclerosis EAS-0783. IMPAIRED INCRETIN SECRETION COMBINED HYPERLIPIDEMIA

IN

PATIENTS

WITH

FAMILIAL

A. Molinea, J. Limaa, X. Sanzb, J. Vidalc, V. Fonollosad, M. Vilardelle a Cardiovascular Risk Unit Internal Medicine Department, Vall d'Hebron University Hospital, Barcelona, Spain; b Internal Medicine Department, Hospital de Barcelona, Barcelona, Spain; c Endocrinology Department, Hospital Clínic, Barcelona, Spain; d Cardiovascular Risk Unit Internal Medicine Department, Vall d’Hebron University Hospital, Barcelona, Spain; e Cardiovascular Risk Unit Internal Medicine Department, Vall d’Hebron University Hospital, Barcelona, Spain

Objectives: Familial combined hyperlipidemia (FCH) is the most frequent lipid genetic disorder in developed countries and is associated with a high incidence of premature cardiovascular disease. FCH is also associated with insulin resistance, abdominal obesity, fatty liver disease, endothelial dysfunction, low-grade inflammation and with the classic lipid phenotype of T2DM, presenting high triglyceride plasma level, decreased high density lipoprotein (HDL) cholesterol and increased amounts of small, dense LDL particles. Furthermore, patients with FCH are at greater risk of developing T2DM. Additionaly, impaired incretin secretion has been observed in diabetic individuals, and in other populations at high risk of developing T2DM, like patients with metabolic syndrome and impaired glucose tolerance. Apart from its insulinotropic effects, GLP-1 and GIP have been related direct and indirectly with lipid metabolism. Incretin secretion has not been previously studied in patients with FCH. The aim of this study was to investigate if patients with FCH have an impaired incretin secretion pattern. Methods: We conducted a case-control study, enrolling 30 patients with known FCH and 14 healthy controls. After the extraction of a basal blood sample, we administered an oral mixed (glucose and lipids) load to both groups, and measured GIP, GLP-1 and GLP-2 concentrations at 30 minutes intervals up to 2 hours after the oral load. Results: There were no statistical differences in age and anthropometric characteristics between both groups. Absolute secretion of GIP and GLP-1 in response to the oral mixed load, calculated as area under the 120 minutes secretion curve, was significantly higher in patients with FCH than in control subjects. There were no significant differences between groups in GLP-2 secretion. Conclusion: Patients with FCH have an impaired incretin secretion pattern, with increased GIP and GLP-1 postprandial plasmatic levels . 08 - Adipose tissue biology EAS-0586. FREE FATTY ACIDS AND ADIPONECTIN IN TUNISIAN PATIENTS WITH METABOLIC SYNDROME

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H. Chahed, N. Gammoudi, S. Ferchichi, A. Miled Biochemistry, Farhat Hached University Hospital, Sousse, Tunisia Objectives: The aim of our study was to evaluate adiponectin and free fatty acid (FFA) in patients with metabolic syndrome (MS). The metabolic syndrome (MS) is a cluster of interrelated common clinical disorders, including hypertension, insulin resistance, glucose intolerance, dyslipidaemia, and obesity. Adiponectin is an adipokine secreted specifically from the adipose tissue. Serum adiponectimia was associated with the risk of MS. Methods: Our study involved 53 patients with MS (mean age 49
14 years) and 55 healthy subjects constituted the control group (mean age 41
10 years). Serum adiponectin were measured with ELISA method (Abcys). FFA was determined by enzymatic colorimetric method at 550 nm (Randox, Antrim, UK). The study was approved by the local Ethics committee Results: Serum adiponectin were significantly lower in patients with MS than control groups (adiponectin: 7. 35
2.20 mg/ml vs 11.12
3.46 mg/ml, p <10-3). While, serum FFA was significantly higher in patients with MS than control groups (FFA: 0.85
0. 51 mmol/l vs 0.42
0. 17 mmol/l, p <10-3). We found that adiponectin was correlated negatively with FFA (r:-0.322; P<10-3) Conclusion: Our data conclude that hypoadiponectemia associated to higher level of FFA were involved in the development of metabolic syndrome in patients. 09 - Hypolidaemic drugs present and future EAS-0738. EFFECT OF TAMOXIFEN, RALOXIFENE AND TOREMIFENE ON LDL RECEPTOR ACTIVITY IN LYMPHOCYTES FROM NORMOLIPIDEMIC AND FAMILIAL HYPERCHOLESTEROLEMIC SUBJECTS D. Gomez-Coronadoa, F. Cerratob, M.E. Fernández-Suáreza, R. Alonsoc, M. Alonsod, C. Vázqueze, P. Mataf, M.A. Lasuncióng a Servicio de Bioquímica-Investigación, Hospital Universitario Ramon y Cajal IRyCIS and CIBEROBN, Madrid, Spain; b Servicio de Bioquímica-Investigación, Hospital Universitario Ramon y Cajal IRyCIS, Madrid, Spain; c Lipid Clinic Servicio de Medicina Interna, IIS-Fundación Jiménez Díaz, Madrid, Spain; d Servicio de Pediatría, Hospital Universitario Ramón y Cajal IRyCIS, Madrid, Spain; e Nutrición Clínica y Obesidad Servicio de Endocrinología y Nutrición, Hospital Universitario Ramón y Cajal IRyCIS and CIBEROBN, Madrid, Spain; f Lipid Clinic Servicio de Madicina Interna, IIS-Fundación Jiménez Díaz, Madrid, Spain; g Servicio de Bioquímica-Investigación, Hospital Universitario Ramón y Cajal IRyCIS and CIBEROBN, Madrid, Spain

Objectives: Selective estrogen receptor modulators (SERMs) are widely prescribed for the treatment and prevention of different disorders and symptoms. These drugs consistently reduce LDL-cholesterol levels, but the mechanism for this effect has not been established. Tamoxifen up-regulates LDL receptor (LDLR) in MOLT-4 cells and its combination with lovastatin causes a synergistic effect. In this study we assessed the effect of tamoxifen, raloxifene and toremifene, and their combinations with lovastatin on LDLR activity in primary lymphocytes from normolipidemic and familial hypercholesterolemic (FH) subjects. Methods: Lymphocytes were isolated from peripheral blood and 1,1'-dioctadecyl-3,3,3,3'-tetramethylindocarbocyanineperchlorate (DiI)-labeled LDL uptake was analyzed by flow cytometry. Results: Tamoxifen, toremifene and raloxifene, in this order, stimulated DiILDL uptake by opposing to the LDL-induced down-regulation of LDLR expression. Lymphocytes were less responsive than MOLT-4 cells to the combination of SERMs and lovastatin, with only tamoxifen consistently showing a synergistic effect with the statin. The increase of lymphocyte LDLR activity by SERMs was not altered by the antiestrogen ICI 182,780, nor is reproduced by 17b-estradiol. However, the active tamoxifen metabolite endoxifen was equally effective as its precursor. In