Cholesterol Forms and Traditional Lipid Profile for Projection of Atherogenic Dyslipidemia: Lipoprotein Subfractions and Erythrocyte Membrane Cholesterol

Atherogenic dyslipidemia characterized by abnormal changes in plasma lipid profile such as low high-density lipoprotein (HDL) and increased triglyceride (TG) levels is strongly associated with atherosclerotic diseases. We aimed to evaluate the levels of pro- and antiatherogenic lipids and erythrocyte membrane cholesterol (EMC) content in normo- and dyslipidemic subjects to investigate whether EMC content could be a useful marker for clinical presentation of atherogenic dyslipidemia. Low-density lipoprotein (LDL), HDL and their subfraction levels and erythrocyte lipid content were determined in 64 normolipidemic (NLs), 42 hypercholesterolemic (HCs) and 42 mixed-type dyslipidemic subjects (MTDs). Plasma atherogenic lipid indices [small–dense LDL (sdLDL)/less-dense HDL (LHDL), TC/HDL-C, TG/HDL-C and Apo B/AI] were higher in MTDs compared to NLs (p < 0.001). The highest sdLDL level was observed in HCs (p < 0.01). Despite a slight increase in EMC level in dyslipidemic subgroups, the difference was not statistically significant. A significant negative correlation, however, was observed between EMC and sdLDL/LHDL in HCs (p < 0.035, r = ?0.386). Receiver operating characteristic curves to predict sdLDL level showed that TG and EMC levels had higher area under curve values compared to other parameters in HCs. We showed that diameters of larger LDL and HDL particles tend to shift toward smaller values in MTDs. Our results suggest that EMC content and TG levels may be a useful predictor for sdLDL level in hypercholesterolemic patients.     

Addition of dietary fat to cholesterol in the diets of LDL receptor knockout mice: effects on plasma insulin, lipoproteins, and atherosclerosis

The factors underlying cardiovascular risk in patients with diabetes have not been clearly elucidated. Efforts to study this in mice have been hindered because the usual atherogenic diets that contain fat and cholesterol also lead to obesity and insulin resistance. We compared plasma glucose, insulin, and atherosclerotic lesion formation in LDL receptor knockout (Ldlr(-/-)) mice fed diets with varying fat and cholesterol content that induced similar lipoprotein profiles. Ldlr(-/-) mice fed a high-fat diet developed obesity, mild hyperglycemia, hyperinsulinemia, and hypertriglyceridemia. Quantitative and qualitative assessments of atherosclerosis were unchanged in diabetic Ldlr(-/-) mice fed a high-fat diet compared with lean nondiabetic control mice after 20 weeks of diet. Although one group of mice fed diets for 40 weeks had larger lesions at the aortic root, this was associated with a more atherogenic lipoprotein profile. The presence of a human aldose reductase transgene had no effect on atherosclerosis in fat-fed Ldlr(-/-) mice with mild diabetes. Our data suggest that when lipoprotein profiles are similar, addition of fat to a cholesterol-rich diet does not increase atherosclerotic lesion formation in Ldlr(-/-) mice.     

Evaluation of the anti-atherogenic potential of chrysin in Wistar rats

Hypercholesterolemia is one of the major risk factors that precipitate coronary heart disease and atherosclerosis. Oxidative stress is believed to contribute to the pathogenesis of hypercholesterolemic atherosclerosis; hence, various antioxidant compounds are being evaluated for potential anti-atherogenic effects. In the present study, the putative anti-atherogenic and antioxidant efficacy of a flavonoid, chrysin, was evaluated in an experimental model of atherosclerosis. In male, albino Wistar rats fed an atherogenic diet for 45 days and treated with saline, significantly higher mean levels of serum lipid profile parameters (total cholesterol, triglycerides, low-density, and very low-density lipoprotein cholesterol), lower mean levels of high-density lipoprotein cholesterol and higher mean serum levels of hepatic marker enzymes (aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, and lactate dehydrogenase) were observed when compared with the levels in rats fed a control diet. In addition, significantly lower mean hepatic levels of lipoprotein lipase, 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase, antioxidant enzymes (catalase, superoxide dismutase, and glutathione peroxidase) and non-enzymatic antioxidants (reduced glutathione, and vitamins C and E), and a significantly higher mean level of hepatic malondialdehyde (MDA) were noted in comparison to the values in control rats. In atherogenic diet-fed rats that received chrysin orally (200 mg/kg b.wt) for 15 days, starting 30 days after the start of the atherogenic diet, significantly lower mean serum levels of lipid profile parameters (except for HDL-cholesterol which was elevated), hepatic marker enzymes, and significantly higher mean hepatic levels of LPL, HMG-CoA reductase, enzymatic, and non-enzymatic antioxidants and significantly lower mean levels of hepatic MDA were noted, compared to the values in atherogenic diet-fed, saline-treated rats. Histopathological studies appeared to suggest the protective effect of chrysin on the hepatic tissue and aorta of atherosclerotic rats. These results suggest that chrysin has anti-atherogenic potential in an experimental setting.     

Lipoprotein subfraction oxidation in acute exercise and ageing

Exercise and ageing can independently increase free radical production that may enhance the susceptibility of LDL to oxidation and create a more atherogenic LDL particle. This investigation was designed to examine exercise and ageing on the susceptibility of LDL subfractions to oxidation. Eleven aged (55?± 4 years) and twelve young (21?± 2 years) participants completed a progressive exercise test to exhaustion and within one week performed a 1?h bout of moderate intensity (65% VO_2max) exercise. Blood was assayed for metabolites associated with lipid composition (total cholesterol, free cholesterol, triglycerides) and lipoprotein susceptibility to oxidation. Exercise increased small density (sdLDL) oxidation, independently of age (p?<?0.05). However, sdLDL oxidation further increased 24?h post exercise in the aged group (p?<?0.05). With regards to the changes in lipid components within LDL, free and total cholesterol and triglycerides in large buoyant (lbLDL) were all elevated 24?h post exercise in aged compared with young (p?<?0.05 for all comparisons). There was a decrease in triglycerides in medium density (mdLDL) 24?h post exercise in the aged group (p?<?0.05). The lipid composition of sdLDL, VLDL, HDL₂, HDL₃ and serum lipid hydroperoxides remained unchanged as a function of exercise and ageing (p?>?0.05). Although regular exercise training is known to be protective against cardiovascular disease (CVD) onset, our data demonstrates that acute exercise can increase sdLDL oxidative susceptibility, and this is independent of age and regardless of a change in LDL lipid composition. However, age seems to be a determining factor with regards the susceptibility of sdLDL to oxidation 24?h following exercise.     

Manzamine A,a marine-derived alkaloid,inhibits accumulation of cholesterol ester in macrophages and suppresses hyperlipidemia and atherosclerosis in vivo

The formation of foam cells in macrophages plays an essential role in the progression of early atherosclerotic lesions and therefore its prevention is considered to be a promising target for the treatment of atherosclerosis. We found that an extract of the marine sponge Acanthostrongylophora ingens inhibited the foam cell formation induced by acetylated low-density lipoprotein (AcLDL) in human monocyte-derived macrophages, as measured based on the accumulation of cholesterol ester (CE). Bioassay-guided purification of inhibitors from the extract afforded manzamines. Manzamine A was the most potent inhibitor of foam cell formation, and also suppressed CE formation in Chinese hamster ovary cells overexpressing acyl-CoA:cholesterol acyl-transferase (ACAT)-1 or ACAT-2. In addition, manzamine A inhibited ACAT activity. Next, we orally administered manzamine A to apolipoprotein E (apoE)-deficient mice for 80 days, and found that total cholesterol, free cholesterol, LDL-cholesterol, and triglyceride levels in serum were significantly reduced and the area of atherosclerotic lesions in the aortic sinus was also substantially diminished. These findings clearly suggest that manzamine A suppresses hyperlipidemia and atherosclerosis in apoE-deficient mice by inhibiting ACAT and is therefore a promising lead compound in the prevention or treatment of atherosclerosis. Although manzamine A has been reported to show several biological activities, this is the first report of a suppressive effect of manzamine A on atherosclerosis in vivo.     

Association of Small Dense LDL Serum Levels and Circulating Monocyte Subsets in Stable Coronary Artery Disease

Objective

Atherosclerosis is considered to be an inflammatory disease in which monocytes and monocyte-derived macrophages play a key role. Circulating monocytes can be divided into three distinct subtypes, namely in classical monocytes (CM; CD14++CD16-), intermediate monocytes (IM; CD14++CD16+) and non-classical monocytes (NCM; CD14+CD16++). Low density lipoprotein particles are heterogeneous in size and density, with small, dense LDL (sdLDL) crucially implicated in atherogenesis. The aim of this study was to examine whether monocyte subsets are associated with sdLDL serum levels.

Methods

We included 90 patients with angiographically documented stable coronary artery disease and determined monocyte subtypes by flow cytometry. sdLDL was measured by an electrophoresis method on polyacrylamide gel.

Results

Patients with sdLDL levels in the highest tertile (sdLDL≥4mg/dL;T3) showed the highest levels of pro-inflammatory NCM (15.2±7% vs. 11.4±6% and 10.9±4%, respectively; p<0.01) when compared with patients in the middle (sdLDL=2-3mg/dL;T2) and lowest tertile (sdLDL=0-1mg/dL;T1). Furthermore, patients in the highest sdLDL tertile showed lower CM levels than patients in the middle and lowest tertile (79.2±8% vs. 83.9±7% and 82.7±5%; p<0.01 for T3 vs. T2+T1). Levels of IM were not related to sdLDL levels (5.6±4% vs. 4.6±3% vs. 6.4±3% for T3, T2 and T1, respectively). In contrast to monocyte subset distribution, levels of circulating pro- and anti-inflammatory markers were not associated with sdLDL levels.

Conclusion

The atherogenic lipoprotein fraction sdLDL is associated with an increase of NCM and a decrease of CM. This could be a new link between lipid metabolism dysregulation, innate immunity and atherosclerosis.     

Atherogenic lipid profile is a feature characteristic of patients with early rheumatoid arthritis: effect of early treatment – a prospective, controlled study

We investigated lipid profiles and lipoprotein modification after immuno-intervention in patients with early rheumatoid arthritis (ERA). Fifty-eight patients with ERA who met the American College of Rheumatology (ACR) criteria were included in the study. These patients had disease durations of less than one year and had not had prior treatment for it. Smokers or patients suffering from diabetes mellitus, hypothyroidism, liver or kidney disease, Cushing's syndrome, obesity, familiar dyslipidemia and those receiving medications affecting lipid metabolism were excluded from the study. Sixty-three healthy volunteers (controls) were also included. Patients were treated with methotrexate and prednisone. Lipid profiles, disease activity for the 28 joint indices score (DAS-28) as well as ACR 50% response criteria were determined for all patients. The mean DAS-28 at disease onset was 5.8 ± 0.9. After a year of therapy, 53 (91.3%) patients achieved the ACR 20% response criteria, while 45 (77.6%) attained the ACR 50% criteria. In addition, a significant decrease in the DAS-28, C-reactive protein (CRP) and erythrocyte sedimentation rate (ESR) were observed. ERA patients exhibited higher serum levels of total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C) and triglycerides, whereas their serum high-density lipoprotein cholesterol (HDL-C) levels were significantly lower compared to controls. As a consequence, the atherogenic ratio of TC/HDL-C as well as that of LDL-C/HDL-C was significantly higher in ERA patients compared to controls. After treatment, a significant reduction of the atherogenic ratio of TC/HDL-C as well as that of LDL-C/HDL-C was observed, a phenomenon primarily due to the increase of serum HDL-C levels. These changes were inversely correlated with laboratory changes, especially CRP and ESR. In conclusion, ERA patients are characterized by an atherogenic lipid profile, which improves after therapy. Thus, early immuno-intervention to control disease activity may reduce the risk of the atherosclerotic process and cardiovascular events in ERA patients.     

Chylomicron remnants are increased in the postprandial state in CD36 deficiency

The clustering of risk factors including dyslipidemia, hyperglycemia, and hypertension is highly atherogenic along with the excess of remnants from triglyceride (TG)-rich lipoproteins. CD36 is involved in the uptake of long-chain fatty acids (LCFAs) in muscles and small intestines. Patients with CD36 deficiency (CD36-D) have postprandial hypertriglyceridemia, insulin resistance, and hypertension. To investigate the underlying mechanism of postprandial hypertriglyceridemia in CD36-D, we analyzed lipoprotein profiles of CD36-D patients and CD36-knockout (CD36-KO) mice after oral fat loading (OFL). In CD36-D patients, plasma triglycerides, apolipoprotein B-48 (apoB-48), free fatty acids (FFAs), and free glycerol levels were much higher after OFL than those of controls, along with increases in chylomicron (CM) remnants and small dense low-density lipoprotein (sdLDL) particles. In CD36-KO mice, lipoproteins smaller than CM in size in plasma and intestinal lymph were markedly increased after OFL and mRNA levels of genes involved in FFA biosynthesis, such as fatty acid binding protein (FABP)-1 and FAS, were significantly increased. These results suggest that CD36-D might increase atherosclerotic risk by enhancing plasma level of CM remnants due to the increased synthesis of lipoproteins smaller than CM in size in the intestine.     

Mast cells promote atherosclerosis by inducing both an atherogenic lipid profile and vascular inflammation

Accumulating in vitro and in vivo studies have proposed a role for mast cells in the pathogenesis of atherosclerosis. Here, we studied the role of mast cells in lipoprotein metabolism, a key element in the atherosclerotic disease. Male mice deficient in low‐density lipoprotein receptors and mast cells on a Western diet for 26 weeks had significantly less atherosclerotic changes both in aortic sinus (55%, P = 0.0009) and in aorta (31%, P = 0.049), as compared to mast cell‐competent littermates. Mast cell‐deficient female mice had significantly less atherosclerotic changes in aortic sinus (43%, P = 0.011). Furthermore, we found a significant positive correlation between the extent of atherosclerosis and the number of adventitial/perivascular mast cells in aortic sinus of mast cell‐competent mice (r = 0.615, P = 0.015). Serum cholesterol and triglyceride levels were significantly lower in both male (63%, P = 0.0005 and 57%, P = 0.004) and female (73%, P = 0.00009 and 54%, P = 0.007) mast cell‐deficient mice, with a concomitant decrease in atherogenic apoB‐containing particles and serum preβ‐high‐density lipoprotein and phospholipid transfer protein activity in both male (69% and 24%) and female (74% and 54%) mast cell‐deficient mice. Serum soluble intercellular adhesion molecule was decreased in both male (32%, P = 0.004) and female (28%, P = 0.003) mast cell‐deficient mice, whereas serum amyloid A was similar between mast cell‐deficient and competent mice. In conclusion, mast cells participate in the pathogenesis of atherosclerosis in ldlr^?/? mice by inducing both an atherogenic lipid profile and vascular inflammation. J. Cell. Biochem. 109: 615–623, 2010. © 2009 Wiley‐Liss, Inc.     

A proteomic study of the apolipoproteins in LDL subclasses in patients with the metabolic syndrome and type 2 diabetes

The exchangeable apolipoproteins present in small, dense LDL (sdLDL) and large, buoyant LDL subclasses were evaluated with a quantitative proteomic approach in patients with the metabolic syndrome and with type 2 diabetes, both with subclinical atherosclerosis and the B LDL phenotype. The analyses included surface-enhanced laser adsorption/ionization, time-of-flight mass spectrometry, and subsequent identification by mass spectrometry or immunoblotting and were carried out in LDL subclasses isolated by ultracentrifugation in deuterium oxide gradients with near physiological salt concentrations. The sdLDLs of both types of patients were enriched in apolipoprotein C-III (apoC-III) and were depleted of apoC-I, apoA-I, and apoE compared with matched healthy controls with the A phenotype. The LDL complexes formed in serum from patients with diabetes with the arterial proteoglycan (PG) versican were also enriched in apoC-III. In addition, there was a significant correlation between the apoC-III content in sdLDL in patients and the apparent affinity of their LDLs for arterial versican. The unique distribution of exchangeable apolipoproteins in the sdLDLs of the patients studied, especially high apoC-III, coupled with the augmented affinity with arterial PGs, may contribute to the strong association of the dyslipidemia of insulin resistance with increased risk for cardiovascular disease.     

血清UA、sdLDL、sST2对急性心肌梗死患者PCI术后无复流的预测价值分析

摘要 目的：探讨血清尿酸（UA）、小而密低密度脂蛋白（sdLDL）、可溶性致癌抑制因子2（sST2）对急性心肌梗死（AMI）患者经皮冠状动脉介入治疗（PCI）术后无复流（NRF）的预测价值。方法：选取2021年1月～2023年1月睢宁县人民医院收治的196例AMI患者为AMI组，根据PCI术后是否发生NRF分为NRF组和血流正常组，另选取同期120名体检健康志愿者为对照组。比较AMI组与对照组血清UA、sdLDL、sST2水平。采用多因素Logistic回归分析AMI患者PCI术后NRF的影响因素，采用受试者工作特征（ROC）曲线分析血清UA、sdLDL、sST2水平对AMI患者PCI术后NRF的预测价值。结果：与对照组比较，AMI组血清UA、sdLDL、sST2水平升高（P＜0.05）。196例AMI患者PCI术后NRF发生率为34.69%，NRF组年龄大于血流正常组，糖尿病比例、肌酸激酶同工酶（CK-MB）、肌钙蛋白I（cTnI）、低密度脂蛋白胆固醇（LDL-C）、UA、sdLDL、sST2水平高于血流正常组（P＜0.05）。多因素Logistic回归分析显示，年龄增加和UA、sdLDL、sST2升高为AMI患者PCI术后NRF的独立危险因素（P＜0.05）。ROC曲线分析显示，血清UA、sdLDL、sST2水平单独和联合预测AMI患者PCI术后NRF的曲线下面积AUC(0.95CI)分别为0.707(0.481～0.934)、0.742(0.513～0.955)、0.737(0.480～0.970)、0.863(0.737～0.960)，联合预测大于单独预测指标。结论：血清UA、sdLDL、sST2水平升高为AMI患者PCI术后NRF的独立危险因素，血清UA、sdLDL、sST2水平联合预测AMI患者PCI术后NRF的价值较高。     

Τhe role of lipoprotein-associated phospholipase A2 in atherosclerosis may depend on its lipoprotein carrier in plasma

Platelet-activating factor (PAF) acetylhydrolase exhibits a Ca²⁺-independent phospholipase A₂ activity and degrades PAF as well as oxidized phospholipids (oxPL). Such phospholipids are accumulated in the artery wall and may play key roles in vascular inflammation and atherosclerosis. PAF-acetylhydrolase in plasma is complexed to lipoproteins; thus it is also referred to as lipoprotein-associated phospholipase A₂ (Lp-PLA₂). Lp-PLA₂ is primarily associated with low-density lipoprotein (LDL), whereas a small proportion of circulating enzyme activity is also associated with high-density lipoprotein (HDL). Τhe majority of the LDL-associated Lp-PLA₂ (LDL-Lp-PLA₂) activity is bound to atherogenic small-dense LDL particles and it is a potential marker of these particles in plasma. The distribution of Lp-PLA₂ between LDL and HDL is altered in various types of dyslipidemias. It can be also influenced by the presence of lipoprotein (a) [Lp(a)] when plasma levels of this lipoprotein exceed 30 mg/dl. Several lines of evidence suggest that the role of plasma Lp-PLA₂ in atherosclerosis may depend on the type of lipoprotein particle with which this enzyme is associated. In this regard, data from large Caucasian population studies have shown an independent association between the plasma Lp-PLA₂ levels (which are mainly influenced by the levels of LDL-Lp-PLA₂) and the risk of future cardiovascular events. On the contrary, several lines of evidence suggest that HDL-associated Lp-PLA₂ may substantially contribute to the HDL antiatherogenic activities. Recent studies have provided evidence that oxPL are preferentially sequestered on Lp(a) thus subjected to degradation by the Lp(a)-associated Lp-PLA₂. These data suggest that Lp(a) may be a potential scavenger of oxPL and provide new insights into the functional role of Lp(a) and the Lp(a)-associated Lp-PLA₂ in normal physiology as well as in inflammation and atherosclerosis. The present review is focused on recent advances concerning the Lp-PLA₂ structural characteristics, the molecular basis of the enzyme association with distinct lipoprotein subspecies, as well as the potential role of Lp-PLA₂ associated with different lipoprotein classes in atherosclerosis and cardiovascular disease.     

Lipoprotein receptors and atherosclerosis

The whole lipoprotein spectrum of human plasma may be divided into atherosclerotic and anti-atherosclerotic lipoproteins. To the first class belong apolipoprotein (apo) B and some apoE-containing lipoproteins of the very-low-density (VLDL), intermediate-density (IDL) and low-density (LDL) lipoprotein fractions. Anti-atherosclerotic lipoproteins are apoA-containing high-density lipoproteins (HDL). Circulating plasma lipoproteins are catabolized mainly by specific cell surface receptors (R) which react with apoB and apoE (B/E-R), for apoE (E-R) or for apoA (HDL-R). Whereas the B/E-R and E-R are responsible for the cellular uptake of lipoproteins and their lipid load by various organs, HDL-R are thought to promote lipid (cholesterol) efflux. There is an additional class of lipoprotein receptors, the so called scavenger-R which are responsible for the removal of altered or degraded lipoproteins for the circulation. Under normal physiological conditions, the concerted action of these receptors warrants efficient lipoprotein turnover and direction into target organs. Derangements of this system, however, may lead to the deposition and accumulation of atherogenic lipids, notably free cholesterol (FC) and cholesteryl esters (CE) in arterial tissue causing atherosclerosis and cardiac death.     

Management of dyslipidemia after coronary artery bypass grafting

The results of serial angiographic studies and intervention trials in patients after coronary artery bypass artery grafting have provided ample evidence that abnormalities of the plasma lipoprotein system are one of the most significant risk factors for a rapid atherosclerotic attrition of saphenous vein grafts. In addition to confirming the well recognized role and contribution of cholesterol-rich LDL or lipoprotein B particles to the progression of atherosclerotic lesions, intervention trials have also provided strong evidence for the atherogenic capacity of some intact and partly delipidized triglyceride-rich very low density lipoprotein and intermediate density lipoprotein (lipoprotein B complex) particles, and the protective effect of some (high density lipoprotein 3) but not all high density lipoprotein particles. Most importantly, those studies have emphasized the need for an early, aggressive treatment of dyslipoproteinemias with pharmacological agents as the most efficient therapeutic approach to delaying, if not preventing, the detrimental effect of atherosclerosis on saphenous vein grafts.     

Anti-Atherogenic Effect of Hydrogen Sulfide by Over-Expression of Cystathionine Gamma-Lyase (CSE) Gene

Hydrogen sulfide (H₂S) is an important gaseous signaling molecule that functions in physiological and pathological conditions, such as atherosclerosis. H₂S dilates vessels and therefore has been suggested as an anti-atherogenic molecule. Since cystathionine gamma-lyase (CSE) enzyme is responsible for producing H₂S in the cardiovascular system, we hypothesized that up-regulation of CSE expression in vivo with preservation of H₂S bioactivity can slow down plaque formation and, can serve as a therapeutic strategy against atherosclerosis. In this study, C57BL/6 wild type mice (WT), ApoE knockout mice (KO) and transgenic ApoE knockout mice overexpressing CSE (Tg/KO) at four weeks of age were weaned. They were then fed with either normal or atherogenic diet for 12 weeks. At week 16, serial plasma lipid levels, body weight, and blood pressure were measured prior to euthanization of the mice and the size of atherosclerotic plaques at their aortic roots was measured. Tg/KO mice showed an increase in endogenous H₂S production in aortic tissue, reduced atherosclerotic plaque sizes and attenuation in plasma lipid profiles. We also showed an up-regulation in plasma glutathionine peroxidase that could indicate reduced oxidative stress. Furthermore, there was an increase in expression of p-p53 and down regulation of inflammatory nuclear factor-kappa B (NF-κB) in aorta. To conclude, alteration of endogenous H₂S by CSE gene activation was associated with reduced atherosclerosis in ApoE-deficient mice. Up-regulation of CSE/H₂S pathway attenuates atherosclerosis and this would be a potential target for therapeutic intervention against its formation.     

Similarity in the distribution of F(2)-isoprostanes in the lipid subfractions of atherosclerotic plaque and in vitro oxidised low density lipoprotein

Oxidation of low density lipoprotein (LDL) in vivo is thought to play a critical role in the initiation of atherosclerosis. F(2)-isoprostanes are compounds resulting from non-enzymatic oxidation of arachidonic acid and elevated levels are present in human atherosclerotic plaque. However, little is known about the formation of F(2)-isoprostanes in plaque lesions or their distribution in lipid subclasses. Given that LDL and tissue lipid subfractions (such as phospholipids, cholesterol esters and triglycerides) all contain significant levels of arachidonic acid, the aim of this study was to examine the relative distribution of F(2)-isoprostanes in the different lipid fractions of LDL oxidised in vitro, and compare this to the distribution in atherosclerotic plaque. The results reveal that while the majority of F(2)-isoprostanes are present in the phospholipid or surface lipid fractions, the core lipids (cholesterol esters/triglycerides) contribute at least 10% of the total F(2)-isoprostanes in both LDL oxidised in vitro and human atherosclerotic plaque. The remarkably similar profiles between the oxidised LDL and advanced atherosclerotic plaque suggests oxidation in vivo, is predominantly via non-enzymatic processes directed towards the surface lipids.     

Endotoxin-lipoprotein complex formation as a factor in atherogenesis: associations with hyperlipidemia and with lecithin:cholesterol acyltransferase activity

A potential role of endotoxin–lipoprotein (bacterial lipopolysaccharide–lipoprotein, LPS–LP) complex formation as a pathogenic factor for atherosclerosis has not been studied yet. The aim of this study was to test the hypothesis that in endotoxinemia in humans hyperlipidemia associated with atherosclerosis development can favor an excessive LPS–LP complex formation, and endotoxin presented in blood can inhibit lecithin:cholesterol acyltransferase (LCAT), one of the key enzymes of reverse cholesterol transport. Endotoxin-binding capacity of lipoproteins (LP) in patients with normolipidemia and hyperlipidemia types IIa and IV was estimated from label incorporation into different LP fractions isolated by means of sequential ultracentrifugation following serum preincubation with Salmonella minnesota R595 ¹²⁵I-labeled LPS. The effect of varied concentrations of S. minnesota R595 LPS on LCAT activity was evaluated from the overall esterifying activity of serum using [1,2-³H₂]cholesterol-labeled substrate. The elevation of low density LP (LDL) and very low density LP (VLDL) contents in blood serum in hyperlipidemia types IIa and IV, respectively, resulted in significant elevation of LPS binding to these fractions. LPS added to the blood serum leads to the dose-dependent decrease in LCAT activity. The revealed phenomena of elevated LPS binding to atherogenic LP fractions in hypercholesterolemia and endotoxin-induced LCAT inhibition suggest the pathogenic role of LPS–LP complexes in atherogenesis.     

Intermedin ameliorates atherosclerosis in apoE null mice by modifying lipid profiles

Intermedin (IMD) is a recently discovered vasodilator peptide. We studied the role of IMD in the pathogenesis of atherosclerosis by investigating the ability of exogenous IMD to alter lipid profiles and ameliorate the development of atherogenic-diet induced atherosclerosis in ApoE−/− mice. Ten of eight-week-old male C57BL/6J mice were as control. Thirty of eight-week-old male ApoE−/− mice were fed with an atherogenic diet for 18 weeks. After feeding atherogenic diet for 12 weeks, the mice were equally and randomly divided into three groups. Normal saline was given in group A and C57BL/6J mice. Intermedin was given by mini osmotic pumps at the dosage of 100 ng/kg/h and 500 ng/kg/h in group B and group C respectively. After the treatment of IMD for 6 weeks, aortic ultrasonography of group C showed that IMD prevented the progression of atherosclerotic lesions and the increase of wall thickness in the aorta. Oil-red-O staining of the entire aorta and the atherosclerotic aortic root section showed 2 folds decrease atherogenic plaque (p < 0.05). Serum lipid profiles were measured, compared with the group A, in group C TC and LDL-C levels were decreased by 86.32% and 89.68%, respectively (both p < 0.05), meanwhile, HDL-C level was significantly increased by 74.82% (p < 0.05). These data indicate that exogenous administration of IMD could prevent the progression of atherosclerotic plaque. The possible underlying mechanisms may relate to the improvement of lipid profiles.     

A simple and sensitive method for lipoprotein and lipids profiles analysis of individual micro-liter scale serum samples

A simple and sensitive method to determine lipoprotein and lipids profiles in micro-liter scale individual serum sample is not presently available. Traditional lipoprotein separation techniques either by ultra-centrifugation or by liquid chromatography methods have their disadvantages in both lipoprotein separation and lipids component quantification. In this study we used small volume needing size-exclusion fast protein liquid chromatography to separate different lipoprotein subclasses in 50μL serum. And lipids contents, such as cholesterol, cholesterol ester and triacylglycerol, were measured by using two different fluorescence-based lipid detection methods. With this method, very low density lipoprotein, low density lipoprotein and high density lipoprotein could be easily separated, and follow-up lipid detection was completed by simple kinds of reactions. Serum lipoprotein and lipids profiling from C57BL/6 mice (n=5) and human (n=5) were analyzed. The elution profiles of five individuals were highly reproducible, and there were lipoprotein and lipids distribution variations between C57BL/6 mice and human beings. In conclusion, this method which combined small volume needing size-exclusion fast protein liquid chromatography and fluorescence-based lipids measurement, provided a simple, efficient, integrity and reproducible procedure for determining serum lipoprotein and lipids profiles in micro-liter scale levels. It becomes possible that determination of lipoprotein profiles and gaining information of lipids in different lipoproteins can be accomplished simultaneously.     

Daily melatonin supplementation in mice increases atherosclerosis in proximal aorta

Considerable evidence supports the hypothesis that LDL oxidation plays an important role in atherosclerosis. Even though high melatonin doses inhibit LDL oxidation in vitro, the effect of melatonin on atherosclerosis has never been studied. We have demonstrated that the feeding of hypercholesterolemic mice with an atherogenic diet supplemented with melatonin highly increases the surface of atherosclerotic lesions in the proximal aorta. These observations occur without detectable lipidic or glucidic phenotype alteration. Melatonin treatment increased highly the sensitivity of atherogenic lipoprotein to Cu(2+) and gamma-radiolysis generated oxyradical ex vivo oxidation during the fasting period. Moreover, these altered lipoproteins were less recognized by the LDL receptor metabolic pathway of murine fibroblasts while they transferred many more cholesteryl esters to murine macrophages. This study suggests that caution should be taken as regards high melatonin dosage in hypercholesterolemic patients.