RESUMEN
It was previously reported that a protein-free microemulsion (LDE) with structure roughly resembling that of the lipid portion of low density lipoprotein (LDL) was presumably taken up by LDL receptors when injected into the bloodstream. In contact with plasma, LDE acquires apolipoproteins (apo) including apo E that would be the ligand for receptor binding. Currently, apo were associated to LDE by incubation with high density lipoprotein (HDL). LDE-apo uptake by mononuclear cells showed a saturation kinetics, with an apparent K(m) of 13.1 ng protein/mL. LDE-apo is able to displace LDL uptake by mononuclear cells with a Ki of 11.5 ng protein/mL. LDE without apo is, however, unable to displace LDL. The uptake of 14C-HDL is not dislocated by increasing amounts of LDE-apo, indicating that HDL and LDE-apo do not bind to the same receptor sites. In human hyperlipidemias, LDE labeled with 14C-cholesteryl ester behaved kinetically as expected for native LDL. LDE plasma disappearance curve obtained from eight hypercholesterolemic patients was markedly slower than that from 10 control normolipidemic subjects [fractional clearance rate (FCR) = 0.02 +/- 0.01 and 0.12 +/- 0.04 h-1, respectively; P < 0.0001]. On the other hand, in four severely hypertriglyceridemic patients, LDE FCR was not significantly different from the controls (0.07 +/- 0.03 h-1). These results suggest that LDE can be a useful device to study lipoprotein metabolism.
Asunto(s)
Emulsiones/farmacocinética , Hiperlipidemias/tratamiento farmacológico , Lipoproteínas/sangre , Lipoproteínas/farmacocinética , Receptores de LDL/metabolismo , Adulto , Anciano , Apolipoproteínas/farmacocinética , Unión Competitiva , Radioisótopos de Carbono , Ésteres del Colesterol/farmacocinética , Emulsiones/química , Femenino , Humanos , Hipercolesterolemia/tratamiento farmacológico , Hipercolesterolemia/metabolismo , Hiperlipidemias/metabolismo , Hipertrigliceridemia/tratamiento farmacológico , Hipertrigliceridemia/metabolismo , Cinética , Leucocitos Mononucleares/efectos de los fármacos , Leucocitos Mononucleares/metabolismo , Lípidos/sangre , Lipoproteínas/química , Lipoproteínas LDL/metabolismo , Lipoproteínas LDL/farmacocinética , Masculino , Persona de Mediana EdadRESUMEN
1. The transfer of lipids that constitute the surface of lipoprotein particles, phospholipids and unesterified cholesterol, from chylomicrons and VLDL to other lipoproteins, mainly HDL, was examined. 2. Emulsions known to mimic the metabolism of chylomicrons labeled with [3H]-phosphatidylcholine, [14C]-cholesterol or [3H]-triolein were injected through a cannula implanted into the carotid artery of male Wistar rats weighing 280-350 g. Plasma clearance of the radioactively labeled emulsion lipid constituents and transfer of surface lipids from the emulsions to native HDL and LDL were measured in plasma samples collected at 2-min intervals during 10 min. 3. The transfer was measured in rats with alloxan-induced diabetes (single intraperitoneal dose, 140 mg/kg body weight) or with propylthiouracil-induced hypothyroidism (0.1% v/v in drinking water for 30 days), or given ethanol (20% in drinking water) for a period of 30 days, and in control rats. 4. The entry of emulsion phospholipids into the HDL fraction was not affected by the different treatments (controls: 9.3 +/- 0.6% of injected radioactivity, N = 8; diabetes: 12.5 +/- 2.4%, N = 9; hypothyroidism: 10.9 +/- 0.9%, N = 13; ethanol: 7.8 +/- 0.9%, N = 5). However, phospholipid transfer to the LDL fraction was increased in diabetes (10.0 +/- 2.3%) and hypothyroidism (12.1 +/- 1.3%) compared to controls (6.7 +/- 0.9%). Transfer of unesterified cholesterol from the emulsions to LDL and HDL was increased in both diabetic and hypothyroid rats (controls, LDL: 1.6 +/- 0.6%, HDL: 2.5 +/- 0.6, N = 5; diabetes, LDL: 5.3 +/- 1.2, HDL: 8.4 +/- 2.1 N = 5; hypothyroidism, LDL: 4.0 +/- 0.6, HDL: 3.6 +/- 0.4, N = 8). In ethanol-treated rats, transfer of surface lipids was similar to controls.
Asunto(s)
Colesterol/metabolismo , Lipoproteínas HDL/metabolismo , Fosfolípidos/metabolismo , Triglicéridos/metabolismo , Aloxano , Animales , Quilomicrones/metabolismo , Diabetes Mellitus Experimental/inducido químicamente , Etanol , Hipotiroidismo/inducido químicamente , Lipólisis , Lipoproteínas LDL/biosíntesis , Masculino , Propiltiouracilo , Ratas , Ratas WistarRESUMEN
1. The transfer of lipids that constitute the surface of lipoprotein particles, phospholipids and unesterified cholesterol, from chylomicrons and VLDL to other lipoproteins, mainly HDL, was examined. 2. Emulsions known to mimic the metabolism of chylomicrons labeled with [3H]-phosphatidylcholine, [14C]-cholesterol or [3H]-triolein were injected through a cannula implanted into the carotid artery of male Wistar rats weighing 280-350 g. Plasma clearance of the radioactively labeled emulsion lipid constituents and transfer of surface lipids from the emulsions to native HDL and LDL were measured in plasma samples collected at 2-min intervals during 10 min. 3. The transfer was measured in rats with alloxan-induced diabetes (single intraperitoneal dose, 140 mg/kg body weight) or with propylthiouracil-induced hypothyroidism (0.1 per cent v/v in drinking water for 30 days), or given ethanol (20 per cent in drinking water) for a period of 30 days, and in control rats. 4. The entry of emulsion phospholipids into the HDL fraction was not affected by the different treatments (controls: 9.3 +/- 0.6 per cent of injected radioactivity, N = 8; diabetes: 12.5 +/- 2.4 per cent , N = 9; hypothyroidism: 10.9 +/- 0.9 per cent , N = 13; ethanol: 7.8 +/- 0.9 per cent , N = 5). However, phospholipid transfer to the LDL fraction was increased in diabetes (10.0 +/- 2.3 per cent ) and hypothyroidism (12.1 +/- 1.3 per cent ) compared to controls (6.7 +/- 0.9 per cent ). Transfer of unesterified cholesterol from the emulsions to LDL and HDL was increased in both diabetic and hypothyroid rats (controls, LDL: 1.6 +/- 0.6 per cent , HDL: 2.5 +/- 0.6, N = 5; diabetes, LDL: 5.3 +/- 1.2, HDL: 8.4 +/- 2.1 N = 5; hypothyroidism, LDL: 4.0 +/- 0.6, HDL: 3.6 +/- 0.4, N = 8). In ethanol-treated rats, transfer of surface lipids was similar to controls
Asunto(s)
Animales , Masculino , Ratas , Colesterol/metabolismo , Lipoproteínas HDL/metabolismo , Fosfolípidos/metabolismo , Triglicéridos/metabolismo , Aloxano , Diabetes Mellitus Experimental/inducido químicamente , Etanol , Hipotiroidismo/inducido químicamente , Lipólisis , Lipoproteínas LDL/biosíntesis , Propiltiouracilo , Quilomicrones/metabolismo , Ratas WistarRESUMEN
The influence of lipolytic mechanisms on the transfer of phospholipids and unesterified cholesterol from artificial emulsions, serving as chylomicron models to other plasma lipoproteins, mainly high density lipoproteins (HDL) were tested in vivo. The emulsions labeled with radioactive lipids were injected into the bloodstream of rats (controls) and the results were compared with those obtained from rats that had previously been treated with Triton WR 1339 or heparin. Plasma clearance and the distribution of cholesteryl esters, phospholipids and unesterified cholesterol in the different plasma lipoprotein fractions were then determined. Whereas virtually no cholesteryl esters were found in d greater than 1.006 g/mL density fraction of the three experimental groups, 2.8 +/- 1.3% of the injected phospholipids were in the 1.063-1.210 g/L density fraction of the Triton treated rats, and 12.6 +/- 5.4% of the heparin treated rats, as compared to 10.1 +/- 1.7% in controls. This indicates that lipolysis directly influences phospholipid transfer to HDL. In contrast, free-cholesterol transfer to HDL, besides being less pronounced than phospholipid transfer, was enhanced by Triton and diminished by heparin, indicating that lipolytic mechanisms were not important determinants in this process.