Expression of fat mobilizing genes in human epicardial adipose tissue.

BACKGROUND: Epicardial adipose tissue (EAT) mass correlates with metabolic syndrome and coronary artery disease (CAD). However, little is known about the expression of genes involved in triglyceride (TG) storage and mobilization in EAT. We therefore analyzed the expression of genes involved in fat mobilization in EAT in comparison to subcutaneous abdominal adipose tissue (AAT) in CAD patients and in controls. METHODS: EAT and AAT were obtained during coronary artery bypass graft (CABG) surgery from 16 CAD patients and from 14 non-CAD patients presenting for valve surgery. The state of atherosclerosis was assessed by angiography. RNA from tissues were extracted, reversibly transcribed and quantified by real time polymerase chain reaction (RT-PCR). The following genes were analyzed: perilipin-1 and -5 (PLIN1, PLIN5), lipoprotein lipase (LPL), hormone sensitive lipase (HSL), adipose triglyceride lipase (ATGL), comparative gene identification-58 (CIG-58), angiopoietin like protein 4 (ANGPTL4), in addition to interleukine-6 (IL-6), leptin (LEP) and adiponectin (ADPN). RESULTS: A significant expression of all listed genes could be observed in EAT. The relative expression pattern of the 10 genes in EAT was comparable to the expression in AAT, yet there was a significantly higher overall expression in AAT. The expression of the listed genes was not different between CAD patients and controls. CONCLUSION: It is suggested that the postulated difference in EAT volume between CAD patients and non-CAD patients is not caused by a differential mRNA expression of fat mobilizing genes. Further work on protein levels and enzyme activities will be necessary to get a complete picture.

Plasma delipidation process induces rapid regression of atherosclerosis and mobilisation of adipose tissue.

Cholesterol is a major component of atherosclerotic plaques. Cholesterol accumulation within the arterial intima and atherosclerotic plaques is determined by the difference of cellular cholesterol synthesis and/or influx from apo B-containing lipoproteins and cholesterol efflux. In humans, apo A-1 Milano infusion has led to rapid regression of atherosclerosis in coronary arteries. We hypothesised that a multifunctional plasma delipidation process (PDP) would lead to rapid regression of experimental atherosclerosis and probably impact on adipose tissue lipids. In hyperlipidemic animals, the plasma concentrations of cholesterol, triglyceride and phospholipid were, respectively, 6-, 157-, and 18-fold higher than control animals, which consequently resulted in atherosclerosis. PDP consisted of delipidation of plasma with a mixture of butanol-diisopropyl ether (DIPE). PDP removed considerably more lipid from the hyperlipidemic animals than in normolipidemic animals. PDP treatment of hyperlipidemic animals markedly reduced intensity of lipid staining materials in the arterial wall and led to dramatic reduction of lipid in the adipose tissue. Five PDP treatments increased apolipoprotein A1 concentrations in all animals. Biochemical and hematological parameters were unaffected during PDP treatment. These results show that five PDP treatments led to marked reduction in avian atherosclerosis and removal of lipid from adipose tissue. PDP is a highly effective method for rapid regression of atherosclerosis.

Effects of simvastatin on blood lipids, vitamin E, coenzyme Q10 levels and left ventricular function in humans.

BACKGROUND: As statin therapy has been reported to reduce antioxidants such as vitamin E and coenzyme Q10 and there are indications that this reduction may cause impairment of left ventricular function (LVF), we studied the influence of simvastatin on LVF and serum vitamin E and coenzyme Q10 levels in humans. MATERIAL AND METHODS: We assessed the effect of simvastatin on left ventricular function and coenzyme Q10 levels in 21 (11 male, 10 female) hypercholesterolaemic subjects (mean age = 56 years) with normal LVF, over a period of 6 months. Subjects were re-tested after a 1-month wash-out period (7 months). Echocardiography was performed on all subjects before commencement of simvastatin (20 mg day(-1)), and at 1, 3, 6 and 7 months after initiation of treatment. Fasting blood samples were also collected at these intervals to assess lipids, apoproteins, vitamin E and coenzyme Q10. RESULTS: Serum lipids showed the expected reductions. Plasma vitamin E and coenzyme Q10 levels were reduced by 17 +/- 4% (P < 0.01) and 12 +/- 4% (P < 0.03) at 6 months. However, the coenzyme Q10/LDL-cholesterol ratio and vitamin E/LDL-cholesterol ratio increased significantly. Left ventricular ejection fraction (EF) decreased transiently after 1 month, while no significant change was observed at 3 and 6 months. Other markers of left ventricular function did not change significantly at any time point. CONCLUSION: Despite reduced plasma vitamin E and coenzyme Q10, 20 mg of simvastatin therapy is associated with a significantly increased coenzyme Q10/LDL-cholesterol ratio and vitamin E/LDL-cholesterol ratio. Simvastatin treatment is not associated with impairment in left ventricular systolic or diastolic function in hypercholesterolaemic subjects after 6 months of treatment.

Therapy of hyper-Lp(a).

Lipoprotein (a) [Lp(a)] appears to be one of the most atherogenic lipoproteins. It consists of a low-density lipoprotein (LDL) core in addition to a covalently bound glycoprotein, apolipoprotein (a) [apo(a)]. Apo(a) exists in numerous polymorphic forms. The size polymorphism is mediated by the variable number of kringle-4 Type-II repeats found in apo(a). Plasma Lp(a) levels are determined to more than 90% by genetic factors. Plasma Lp(a) levels in healthy individuals correlate significantly high with apo(a) biosynthesis and not with its catabolism. There are several hormones known to have a strong impact on Lp(a) metabolism. In certain diseases, such as kidney disease, Lp(a) catabolism is impaired leading to up to fivefold elevations. Lp(a) levels rise with age but are otherwise influenced only little by diet and lifestyle. There is no safe and efficient way of treating individuals with elevated plasma Lp(a) concentrations. Most of the lipid-lowering drugs have either no significant influence on Lp(a) or exhibit a variable effect in patients with different forms of primary and secondary hyperlipoproteinemia. There is without doubt a strong need to concentrate on the development of specific medications to selectively target Lp(a) biosynthesis, Lp(a) assembly and Lp(a) catabolism. So far only anabolic steroids were found to drastically reduce Lp(a) plasma levels. This class of substance cannot, of course, be used for treatment of patients with hyper-Lp(a). We recommend that the mechanism of action of these drugs be studied in more detail and that the possibility of synthesizing derivatives which may have a more specific effect on Lp(a) without having any side effects be pursued. Other strategies that may be of use in the development of drugs for treatment of patients with hyper-Lp(a) are discussed in this review.

[Aggressive therapy and combination therapy in severe hyperlipidemia]. / Aggressive Therapie und Kombinationstherapie bei schweren Hyperlipidämien.

In cases of severe hyperlipidemia, we have very effective therapeutic tools nowadays. LDL-apheresis is still the most effective therapy for familial hypercholesterolemia, especially in its homocygous form. In combination with statins, LDL-cholesterol can be lowered by 80%. Most other genetic and secondary hyperlipidemias can effectively be treated by lipid lowering drugs. This aggressive treatment gains more and more on importance in view of the tough recommendations of specific scientific societies. Atorvastatin and simvastatin can lower LDL-cholesterol by 60%. In this report different therapies and combination therapies for hyperlipidemia are discussed.

Urinary apolipoprotein (a) excretion in patients with proteinuria.

Increased plasma lipoprotein (a) (Lp(a)) levels are strongly associated with premature cardiovascular disease and stroke. Recently we, as well as other groups, found that apolipoprotein (a) (apo(a)) fragments appear in the urine of healthy individuals, and that renal transplant patients with impaired renal function excrete fewer apo(a) fragments into their urine compared with controls. As the excretion mode of apo(a) is presently unknown, we determined plasma Lp(a) levels and urinary apo(a) excretion in relation to kidney function in 58 proteinuric patients and 58 healthy controls. For the first time, urinary apo(a) excretion was related to apo(a) isoforms. Plasma Lp(a) values were higher in the proteinuric patients compared with the controls, independent of their renal function. The patients with low-molecular-weight apo(a) isoforms had higher Lp(a) plasma levels, whereas the patients with high-molecular-weight apo(a) isoforms had lower Lp(a) plasma levels. Urinary apo(a) showed a very similar pattern to that of plasma Lp(a), being significantly higher in patients with low-molecular-weight isoforms as compared with patients with high-molecular-weight isoforms. Urinary apo(a) excretion was significantly decreased in the patient group when compared with healthy controls. There was a close correlation (P < 0.001) between the plasma Lp(a) and urinary apo(a) excretion in both the patient group and the control group. Urinary apo(a) excretion did not correlate with protein excretion, creatinine clearance or plasma creatinine levels. We conclude that urinary apo(a) excretion correlates with plasma Lp(a) and Lp(a) isoforms, and that proteinuric patients excrete significantly less apo(a) into their urine than healthy controls, a factor that might contribute to increased plasma Lp(a) levels in these patients.

[Lowering cholesterol 1998. Cholesterol synthesis inhibitors compared]. / Cholesterinsenkung 1998. Cholesterinsynthese-Hemmer im Vergleich.

Large scale primary and secondary prevention trials in recent years have revealed that the effective lipid reducing therapy with statins can reduce mortality of coronary heart disease by up to 30%. For the first time it has become possible to reduce LDL-cholesterol pharmacologically by more than 50%, a reduction that was only achieved by LDL-apheresis so far. Cost-effectiveness is becoming an important issue since this varies widely between patients according to the coronary risk. Treating the patients with the highest coronary risk is most cost effective. Currently, there are six statins on the market. Reduction of LDL-cholesterol is mainly mediated by the induction of LDL-receptor activity in the liver. In addition, some statins at high doses also reduce LDL-cholesterol synthesis. Due to variations in the molecular structure of the active compounds these 6 statins have important pharmacological differences, such as their capacity to reduce plasma triglycerides, their interaction with other drugs. The daily recommended doses of the statins range from 0.1 mg (cerivastatin) to 80 mg (atorvastatin). In this review the differences in the pharmacological and clinical actions of the statins are analyzed.

Decreased urinary apolipoprotein (a) excretion in patients with impaired renal function.

BACKGROUND: Plasma lipoprotein (a) [Lp(a)] levels are elevated in patients with kidney disease and are strongly associated with premature cardiovascular disease and stroke. METHODS: As the kidney is suggested to play an important role in apolipoprotein (a) [apo(a)] catabolism and as apo(a) fragments appear in urine, we determined plasma Lp(a) levels and urinary apo(a) excretion in relation to kidney function in a large cohort of renal patients. A total of 368 renal patients with normal or different degrees of impaired renal function and 163 healthy control subjects matched for age and sex were investigated. Plasma Lp(a) and urinary apo(a) were analysed immunochemically. RESULTS: Renal patients were found to have significantly elevated total cholesterol and low-density lipoprotein (LDL)-C values but lower high-density lipoprotein (HDL)-C values than control subjects. Plasma Lp(a) values were significantly higher only in patients with creatinine clearance < 70 mL min-1. There was a significant correlation between urinary apo(a) and plasma Lp(a) in patients and control subjects. Urinary apo(a) excretion was significantly lower in patients than in control subjects and showed no correlation with urinary protein excretion. CONCLUSION: Although it is unlikely that impaired renal excretion of apo(a) fragments largely contributes to increased plasma Lp(a) levels in patients suffering from impaired kidney function, these data suggest that urinary apo(a) excretion is significantly decreased in renal patients and that this might contribute to increased plasma Lp(a) levels in this patient group.

Lecithin-cholesterol acyltransferase activity in normocholesterolaemic and hypercholesterolaemic roosters: modulation by lipid apheresis.

Lipid apheresis, a recently described procedure for the elimination of lipid but not apolipoproteins from plasma, was applied to normocholesterolaemic and hypercholesterolaemic roosters. Lipid apheresis resulted in an immediate reduction in plasma unesterified cholesterol concentration, which was sustained for 150 min. The reduction in unesterified cholesterol concentration was higher in the normocholesterolaemic animals than in the hypercholesterolaemic animals. Lipid apheresis induced changes in the ratio of plasma unesterified to total cholesterol in normocholesterolamic animals but not in hypercholesterolaemic animals. In hypercholesterolaemic animals, lecithin-cholesterol acyltransferase (LCAT) activity was not affected by lipid apheresis, whereas in normocholesterolaemic animals LCAT activity was acutely reduced for 150 min after lipid apheresis. Saturated LCAT kinetics occurred in the hypercholesterolaemic animals but not in the normocholesterolaemic animals. LCAT obeyed Michaelis-Menten kinetics. After lipid apheresis, there was a pool of unesterified cholesterol that was available as substrate for LCAT to a greater extent in hypercholesterolaemic animals than in normocholesterolaemic animals. These observations may have important implications for lipid apheresis as a treatment for atherosclerosis.

Urinary apo(a) discriminates coronary artery disease patients from controls.

Increased plasma lipoprotein (a) (Lp(a)) levels are associated with premature cardiovascular diseases and stroke. Since Lp(a) immune reactivity is found in urine we compared urinary apolipoprotein (a) (apo(a)) with plasma Lp(a) levels in 116 patients suffering from angiographically proven coronary artery diseases with that of 109 controls. Urinary apo(a) investigated by immuno blotting, revealed a distinct apo(a) fragmentation pattern with molecular weights between 50 and 160 kDa. Apolipoprotein B however was not secreted into urine. Lp(a) and apo(a) were measured by a fluorescence immuno assay. Within single individuals, urinary apo(a) levels correlated significantly with creatinine (Rho, 0.98; P < 0.0005). Medians and 25/75 percentiles of urinary apo(a) in coronary artery disease (CAD) patients were 5.70, 3.25 and 10.35 microg/dl and in controls 2.64, 1.43 and 3.50 microg/dl respectively. At cut-off levels of 30 mg/dl for plasma Lp(a) and 10 microg/dl of urinary apo(a) respectively, both paramenters showed comparable sensitivities (33.8% vs. 26.7%), yet the specificity (76.1% vs. 91.7%) and the positive predictive value (60.0% vs.76.4%) of urinary apo(a) were much higher. In receiver-operating characteristic plots, urinary apo(a) was much more sensitive at high specificities i.e. greater than 60% as compared to Lp(a). Urinary secretion of apo(a) fragments normalized to creatinine is stable in a given individual and significantly associated with coronary artery disease.

LDL-apheresis significantly reduces urinary apo(a) excretion.

Increased plasma Lp(a) is an established risk factor for atherosclerosis. We recently described the presence of apo(a) fragments in urine and the significant correlation between urinary apo(a) concentrations and plasma Lp(a). Here we investigated urinary apo(a) in patients suffering from familial hypercholesterolaemia (FH), treated with LDL apheresis. Before treatment, plasma Lp(a) levels and urinary apo(a) normalized to creatinine were > 2-fold increased in FH patients (P < 0.0001) as compared to controls. LDL-apheresis led to a reduction of plasma Lp(a) by 75% and a concomitant immediate reduction of urinary apo(a) by 45%. We conclude that a steady state condition for urinary apo(a) is rapidly achieved via LDL-apheresis.

Urinary excretion of apo(a) in patients after kidney transplantation.

BACKGROUND: Increased plasma Lipoprotein (a) (Lp(a)) levels are strongly associated with premature cardiovascular disease and stroke. The kidney is purported to play an important role in apo(a) catabolism. Therefore we investigated plasma Lp(a) levels in relation to kidney function and urinary apo(a) excretion. METHODS: One hundred and sixteen kidney transplant patients with normal or impaired renal function and 109 age- and sex-matched healthy controls were investigated. Plasma Lp(a) and urinary apo(a) levels were determined immunochemically and all other parameters were determined by routine laboratory methods. RESULTS: Transplant recipients were found to have significantly elevated total cholesterol and LDL-C values, but equal HDL-C values compared to controls. Plasma Lp(a) values were higher and urinary apo(a) excretion was lower in transplant recipients compared to controls, independent of renal function. When the patient group was subdivided into 'normal' and 'impaired creatinine clearance', only the latter group secreted less apo(a) than normal controls. CONCLUSION: These data suggest that urinary apo(a) excretion is reduced in transplant recipients with impaired excretory graft function, which may contribute to the elevation of plasma Lp(a) levels in these patients.

Urinary excretion of apo(a) fragments. Role in apo(a) catabolism.

The biosynthesis and assembly of lipoprotein(a) [Lp(a)], a marker for atherosclerotic disease, appears to be well understood. However, information is lacking concerning the mode and site of Lp(a) catabolism. Apo(a) is reported to be excreted into the urine. To study the effect of this pathway on the overall catabolism of Lp(a), urinary apo(a) was characterized by immunoblotting. More than 10 distinct apo(a) bands with molecular masses between 30 and 160 kD were observed. Apo(a) fragments were not complexed to apoB. In more than 30 individuals the size of apo(a) bands was comparable irrespective of their apo(a) phenotype, although marked differences in the relative intensities of the bands were observed. Eight batches of 24-hour urine collections collected from one proband at 2-week intervals exhibited a significant correlation between creatinine and apo(a) concentrations as measured by DELFIA (r = .93; P < .01). In 193 healthy volunteers a highly significant correlation was found between urinary apo(a) concentrations normalized to creatinine levels and plasma Lp(a) values (p = 0.659; P < .0001). Of the total plasma apo(a), 0.073%, i.e., 121 micrograms apo(a), was excreted in the form of apo(a) fragments in 24-hour urine samples from 12 healthy volunteers. We conclude that the catabolism of Lp(a) via excretion of apo(a) fragments accounts for < 1% of the daily Lp(a) catabolism.

Lipid apheresis in an animal model causes in vivo changes in lipoprotein electrophoretic patterns.

Lipid apheresis, a new extracorporeal procedure based on plasma delipidation and showing promise as a possible treatment for atherosclerosis, was recently reported for the first time from this laboratory [Cham et al., J Clin Apheresis 10:61-69, 1995]. In the present study lipid apheresis was applied to hypercholesterolemic and normocholesterolemic roosters to examine its effect on plasma lipoprotein particles. This procedure resulted in conspicuous changes in electrophoretic patterns of plasma lipoproteins. The electrophoretic mobilities of all the lipoprotein fractions had changed considerably. Lipid stainable material was present in at least three bands in the alpha-globulin area. In particular, changes in the electrophoretic region of high-density lipoproteins were observed. Lipid apheresis markedly induced the anti-atherogenic pre- beta-high-density lipoproteins. The observed changes induced by lipid apheresis were more pronounced in the hyperlipidemic animals compared with the normocholesterolemic controls. A novel pre-alpha-lipoprotein band was observed soon after lipid apheresis. This lipoprotein band had a density larger than 1.21. At approximately 150 minutes after lipid apheresis, the electrophoretic pattern had almost returned to its original base pattern. Lipid apheresis results in plasma lipoprotein changes which may induce reverse cholesterol transport and shows promise as a possible treatment of atherosclerosis.

Lipid apheresis: an in vivo application of plasma delipidation with organic solvents resulting in acute transient reduction of circulating plasma lipids in animals.

Despite primary and secondary prevention of coronary disease with lowering plasma cholesterol by diet and drug therapy, coronary heart disease remains the major cause of death in Western countries. Low density lipoprotein apheresis had the potential to make a significant impact as it acutely leads to a marked reduction in plasma cholesterol. However, recent preliminary results suggest that low density lipoprotein apheresis may not be more effective in preventing progression of coronary disease than current drug therapy. We have devised a new technique, termed lipid apheresis, which removes cholesterol and triglycerides from plasma but retains the apolipoproteins. This procedure shows great promise in stimulating regression beyond current therapy. Lipid apheresis, a new extracorporeal procedure based on plasma delipidation with the organic solvent mixture butanol-diisopropyl ether, was applied to hypercholesterolemic and normocholesterolemic roosters. Approximately 25% of the calculated blood volume was removed from the animals. The plasma was separated from the blood cells. The plasma was delipidated for 20 min with the organic solvent mixture. The delipidated plasma containing all proteins, including the apolipoproteins and other ionic constituents, was remixed with the blood cells and infused back into the identical donor animals. Analyses of serial blood samples collected from lipid apheresed and sham treated animals up to 16 h after infusion revealed that lipid apheresis caused acute, marked reductions in plasma lipids. The pattern and extent of the plasma levels of cholesterol were different in the hypercholesterolemic animals when compared with normocholesterolemic animals, indicating that a readily extraplasma cholesterol pool in the hypercholesterolemic animals was rapidly mobilized into the plasma pool.(ABSTRACT TRUNCATED AT 250 WORDS)

[Radical therapy of refractory hyperlipidemia: extracorporeal cholesterol elimination]. / Radikaltherapie der refraktären Hyperlipidämie: extrakorporale Cholesterinelimination.

In Austria atherosclerosis related diseases are responsible for the death of more than 50% of the population. There is a linear relationship between lowering cholesterol and the mortality of coronary heart disease. Apart from diet and drug therapy for the treatment of hypercholesterolemia we nowadays have very potent extracorporeal cholesterol lowering therapies. In combination with HMG-CoA reductase inhibitors LDL-cholesterol can be lowered by 80% with these procedures. In particular homozygote familial hypercholesterolemia is an absolute indication for extracorporeal lipid lowering. Refractory heterozygote FH and secondary hypercholesterolemia in combination with other risk factors can also be controlled by extracorporeal cholesterol elimination. Since some of these extracorporeal therapies also improve the hemorheological situation they are also being used to treat peripheral vascular disease and cerebral atherosclerosis.