Intimal Clearance Research Articles

In vivo transfer of lipoprotein(a) into human atherosclerotic carotid arterial intima.

The aim of this study was to compare the atherogenic potential of lipoprotein(a) [Lp(a)] and LDL by measuring the intimal clearance of these two plasma lipoproteins in the atherosclerotic intima of the human carotid artery in vivo. Autologous 131I-Lp(a) and 125I-LDL were mixed and reinjected intravenously 3 hours before elective surgical removal of the arterial intima in four patients. The intimal clearance of Lp(a) and LDL was 229+/-48 and 405+/-127 nL/cm2 per hour, respectively (paired t test; P=.12). The mass accumulation of Lp(a) (114+/-32 ng/cm2 per hour) was on average one 15th that of LDL (paired t test; P=.06), mainly reflecting a low plasma concentration of Lp(a) compared with LDL in the human subjects studied. In accordance with our previous observation in rabbits, there was a positive association between the intimal clearance of LDL and that of Lp(a) (r=.97, P=.03). Accordingly, high plasma levels of Lp(a) may share with LDL the potential for causing lipid accumulation in the arterial intima in humans.

Preferential influx and decreased fractional loss of lipoprotein(a) in atherosclerotic compared with nonlesioned rabbit aorta.

The aim was to investigate the atherogenic potential of lipoprotein(a) (Lp(a)) and to further our understanding of the atherogenic process by measuring rates of transfer into the intima-inner media (i.e., intimal clearance) and rates of loss from the intima-inner media (i.e., fractional loss) of Lp(a) and LDL using cholesterol-fed rabbits with nonlesioned (n = 13) or atherosclerotic aortas (n = 12). In each rabbit, 131I-Lp(a) (or 131I-LDL) was injected intravenously 26 h before and 125I-Lp(a) (or 125I-LDL) 3 h before the aorta was removed and divided into six consecutive segments of similar size. The intimal clearance of Lp(a) and LDL was similar and markedly increased in atherosclerotic compared with nonlesioned aortas (ANOVA, effect of atherosclerosis: P < 0.0001). Fractional losses of labeled Lp(a) and labeled LDL in atherosclerotic aorta were on average 25 and 43%, respectively, of that in nonlesioned aortas (ANOVA, effect of atherosclerosis: P < 0.0001). Fractional loss of Lp(a) was 73% of that of LDL (ANOVA, effect of type of lipoprotein: P = 0.07). These data suggest that the development of atherosclerosis is associated with increased influx as well as decreased fractional loss of Lp(a) and LDL from the intima. Accordingly, Lp(a) may share with LDL the potential for causing atherosclerosis.

Transfer of lipoprotein(a) and LDL into aortic intima in normal and in cholesterol-fed rabbits.

To study the relative atherogenic potential of lipoprotein(a) [Lp(a)], the transfer of Lp(a) and LDL into the arterial wall was compared in normal rabbits, cholesterol-fed rabbits, and normal rabbits in which the plasma concentration of Lp(a) before injection of labeled lipoproteins was increased by an intravenous mass injection of 45 mg Lp(a). Aorta was removed either 60 minutes or 180 minutes after intravenous injection of a mixed preparation of human 125I-Lp(a) and 131I-LDL; intimal clearance was calculated as radioactivity in aortic intima/inner media divided by the average concentration of the appropriate radioactivity in plasma and by the length of the exposure time. The intimal clearance of labeled Lp(a) and LDL in the aortic arch after 60 minutes of exposure was 87 +/- 9 and 47 +/- 7 nL.cm-2.h-1 (n = 9) in normal rabbits and 82 +/- 14 and 62 +/- 10 nL.cm-2.h-1 (n = 10) in cholesterol-fed rabbits; after 180 minutes of exposure, the intimal clearance of labeled Lp(a) and LDL was 62 +/- 14 and 84 +/- 21 nL.cm-2.h-1 (n = 6) and 30 +/- 6 and 47 +/- 12 nL.cm-2.h-1 (n = 4) in cholesterol-fed and Lp(a)-injected rabbits, respectively. Linear regression analysis showed positive associations between intimal clearance of the two lipoproteins in all four groups of rabbits in the aortic arch, the thoracic aorta, and the abdominal aorta. Aortic immunoreactivity of human apolipoprotein(a) was detected in the intima in association with fatty streak lesions, predominantly within the cytoplasm of foam cells. These results suggest that Lp(a) is transferred into the aortic intima by a mechanism similar to that for LDL and that Lp(a) can be taken up by intimal foam cells; however, Lp(a) and LDL may be metabolized differently upon entrance into the arterial wall.

Accelerated cholesterol accumulation in homologous arterial transplants in cholesterol-fed rabbits. A surgical model to study transplantation atherosclerosis.

Accelerated coronary artery disease has become a major complication to heart transplantation in humans. Therefore, we have developed a surgical model in the rabbit, with transplantation of the thoracic aorta as a bypass graft onto the abdominal aorta of another rabbit. The model permits the study of cholesterol metabolism in transplanted arteries. The graft did not accumulate cholesterol for as long as 298 days, provided that the rabbits were normocholesterolemic, i.e., with plasma cholesterol levels of 0.3-0.7 mmol/l. However, after a few weeks of cholesterol feeding resulting in plasma cholesterol levels of 2-5 mmol/l, the homologous graft accumulated cholesterol compared with intact aortic tissue in the rabbits and also compared with autologous aortic grafts. The intimal clearance of plasma cholesteryl ester, mainly high density lipoprotein cholesteryl ester, in the luminal layer of the aortic graft was 60-150 nl x cm-2 x hr-1 1-2 hours after transplantation. The intimal clearance in the corresponding intact thoracic aorta of the recipient animal was 5-20 nl x cm-2 x hr-1. The values were 1,500-3,000 nl x cm-2 x hr-1 51-298 days after transplantation, while the intimal clearance of the rabbit's own aorta remained unchanged. A pronounced increase in plasma lipoprotein permeability is thus an early event in transplanted arteries. It results in a higher cholesteryl ester influx that leads to cholesterol accumulation in the artery, but only if the rabbits are fed a cholesterol-enriched diet. This rabbit model may be useful in the search for interventional measures to prevent or diminish the accelerated coronary artery disease in transplanted hearts in humans.

Influx in vivo of low density, intermediate density, and very low density lipoproteins into aortic intimas of genetically hyperlipidemic rabbits. Roles of plasma concentrations, extent of aortic lesion, and lipoprotein particle size as determinants.

To compare the atherogenic potential of low density lipoprotein (LDL), intermediate density lipoprotein (IDL), and very low density lipoprotein (VLDL) under conditions where plasma levels of these lipoproteins are elevated, the influx of cholesterol in these lipoproteins into the aortic intima was measured in vivo in genetically hyperlipidemic rabbits from the St. Thomas's Hospital strain, an animal model that shares many of the features of the human disorder familial combined hyperlipidemia. Univariate linear regression showed that the arterial influx of LDL cholesterol (n = 25), IDL cholesterol (n = 14), and VLDL cholesterol (n = 10) was positively and linearly associated with plasma concentrations of LDL cholesterol in the range 0.2-6.4 mmol/l, of IDL cholesterol in the range 0.1-7.0 mmol/l, and of VLDL cholesterol in the range 0.7-8.5 mmol/l, respectively, and also with the extent of lesions in the arterial intima in the range 0-100% of the surface area. Multiple linear regression suggested that the arterial influx of LDL, IDL, and VLDL cholesterol was linearly dependent on plasma concentration, independent of lesion size. Furthermore, it appeared that the arterial influx of the three lipoproteins was linearly dependent on the extent of the lesions, independent of lipoprotein concentration. When influx was normalized for plasma concentration (intimal clearance) and for lesion size (compared within the same aorta), the intimal clearance of the larger IDL and VLDL particles was 15-35% less than that of the smaller LDL particles. These findings suggest that the quantitatively most important mechanism for transfer of plasma lipoproteins into the arterial intima involves nonspecific molecular sieving and that at elevated plasma levels, IDL and VLDL share with LDL the potential for causing atherosclerosis.

Quantitative studies of transfer in vivo of low density, Sf 12-60, and Sf 60-400 lipoproteins between plasma and arterial intima in humans.

To assess the potential of various plasma lipoprotein classes to contribute to the lipid content of the arterial intima, influx and efflux of these plasma lipoprotein fractions into and from the intima of human carotid arteries were measured in vivo. While low density lipoprotein (LDL) is known to transfer from plasma into the arterial wall, there is less information on the atherogenic potential of lipoproteins of intermediate density (Sf 12-60) or of very low density (Sf 60-400). Aliquots of the same lipoprotein (LDL, Sf 12-60 lipoprotein particles, or Sf 60-400 lipoprotein particles) iodinated with iodine-125 and iodine-131 were injected intravenously 18-29 hours and 3-6 hours, respectively, before elective surgical removal of atheromatous arterial tissue, and the intimal clearance of lipoproteins, lipoprotein influx, and fractional loss of newly entered lipoproteins were calculated. Intimal clearance of Sf 60-400 particles was not detectable (less than 0.3 microliter x hr-1 x cm-2), whereas the average value for both LDL and Sf 12-60 lipoprotein particles was 0.9 microliter x hr-1 x cm-2. Since the fractional loss of newly entered LDL and Sf 12-60 lipoprotein particles was also similar, the results suggest similar modes of entry and exit for these two particles. However, due to lower plasma concentrations of Sf 12-60 lipoproteins as compared with LDL, the mass influx of cholesterol in the Sf 12-60 particles was on the order of one 10th of that in LDL, and that of apolipoprotein B was about one 20th.(ABSTRACT TRUNCATED AT 250 WORDS)

Different efflux pathways for high and low density lipoproteins from porcine aortic intima.

To study the efflux of high (HDL) and low (LDL) density lipoproteins from the arterial wall in vivo, a surgical model in pigs was used. An isolated segment of the lesion-free thoracic aorta was pulse labeled from the lumen of the artery with 3H-cholesteryl ester labeled HDL and 14C-cholesteryl ester labeled LDL. Subsequently, the labeled aortic segment was exposed to cold chase in vivo. The transfer of HDL cholesteryl ester from plasma into intima expressed as intimal clearance was three to seven times greater than that of LDL cholesteryl ester. At least 50%, but possibly as much as 95%, of the HDL cholesteryl ester that entered the arterial intima during a period of 4 hours penetrated the arterial wall beyond the internal elastic lamina. In contrast, less than 15% of the LDL cholesteryl ester that entered the arterial intima in the same period penetrated beyond the luminal layer. After 24 hours of cold chase in vivo, more than 80% of both labeled HDL esterified cholesterol and labeled LDL esterified cholesterol had disappeared from the arterial wall. Transmural profiles after 9 hours of cold chase showed that labeled HDL was present throughout the entire arterial wall, whereas labeled LDL in quantitative amounts was present only in the luminal layer. The results suggest that the most important efflux route for HDL esterified cholesterol is through the vasa vasorum and lymphatics in the outer media and adventitia, whereas LDL esterified cholesterol predominantly leaves intima via the lumen of the artery.

Large lipoproteins are excluded from the arterial wall in diabetic cholesterol-fed rabbits

In diabetic hypercholesterolemic rabbits at plasma triglyceride concentrations of approximately 5000 mg/dl, 55% of plasma cholesterol (1400 mg/dl) was in lipoproteins with diameters larger than 75 nm (Sf greater than 400), 40% in smaller very low density and intermediate density lipoproteins, 4% in low density lipoproteins, and 1% in high density lipoproteins. Specific intimal clearance (nl/h.mg aortic cholesterol) of the giant Sf greater than 400 lipoproteins was about 4% of that of the low density lipoproteins. The data suggest that even very low density lipoproteins with diameters smaller than 75 nm were practically excluded from entering the arterial wall. Specific intimal clearance of low density lipoproteins in hypertriglyceridemic, diabetic cholesterol-fed rabbits was similar to that in normal cholesterol-fed rabbits, but low density lipoprotein concentrations in diabetic rabbits were low. Thus, at plasma triglyceride concentrations of approximately 5000 mg/dl, only 5% of plasma cholesterol may be readily available for infiltration of arteries. These results add further support to the hypothesis that hypertriglyceridemic, diabetic cholesterol-fed rabbits are protected against atherogenesis because the major part of plasma cholesterol is carried in large lipoproteins to which the artery is not very permeable.

In vivo fluxes of plasma cholesterol, phosphatidylcholine and protein into mini-pig aortic and pulmonary segments

Levels of radioactive cholesterol found in the arterial wall after a few hours' in vivo exposure to plasma containing labelled cholesterol are so small that the contaminating plasma in the tissue may contribute significantly to the radioactivity measured in the wall. By the simultaneous use of [ 3H]- and [ 14C]cholesterol the contamination was found to be less than 15 nl of plasma/cm 2 of intimal surface of the thoracic aorta in mini-pigs, and this contamination accounted for less than 5% of the total radioactivity found in the intima—media tissue 7 h after the injection of serum with bioincorporated labelled cholesterol. The radioactivity in the arterial wall divided by the area below the plasma radioactivity versus the time curve was named the intimal clearance. For a particular mini-pig 2 nearly identical estimates were obtained for the intimal clearance of labelled plasma cholesterol, using an influx period of about 4 h for [ 3H]cholesterol and 7 h for [ l4C]cholesterol. This indicates a negligible loss of labelled cholesterol from the wall during a 7-h period. The intimal clearances as calculated for labelled plasma cholesterol were about 500, 200, 100 and 50 nl cm −2 h −1 for pulmonary trunk, aortic arch, proximal and distal thoracic aorta, respectively. The concomitantly measured intimal clearances of plasma [ 32P]phosphatidylcholine were about 1.1 times those for plasma cholesterol for the 4 segments. This suggests that the appearance of labelled plasma phosphatidylcholine and of cholesterol in the intima—media is essentially a consequence of a lipoprotein influx. Similarly, the intimal clearances of plasma [ 35S]albumin for the same 4 segments were about 1.5 times those of the labelled lipids, suggesting a non-specific mechanism for the transfer of plasma lipoproteins into the artery. The intimal clearances of [ 35S]albumin were within the same order of magnitude as capillary clearance values for albumin reported for mammalian tissues.

The in vivo transfer of free and esterified cholesterol from plasma into the arterial wall of hypercholesterolemic rabbits

With a new technique based on the simultaneous use of 3H and 14C-cholesterol, the fluxes of free and esterified cholesterol into the thoracic aorta of the hypercholesterolemic rabbits have been measured. By additional use of 3H-leucine as tracer for apoproteins, it was shown that esterified cholesterol in plasma enters the arterial tissue mainly as part of a lipoprotein influx. In contrast, only between 20 and 90% of the influx of free cholesterol could be accounted for by an influx of plasma lipoproteins. The excess probably represents an exchange of free cholesterol between the plasma lipoproteins and the intimal surface of the artery. Some 5-40% of the esterified cholesterol which had entered the artery during a 3-6 hours period was hydrolyzed in the artery during that period, whereas only negligible amounts of newly-entered free cholesterol was esterified. The arterial influx of very low, low and high density lipoproteins or albumin was determined in the same animal. The influx of each fraction was expressed as an intimal clearance, i.e., the influx divided by its plasma concentration. The intimal clearances of these macromolecules decreased linearly with the logarithm of their diameters. This suggests that the arterial influx of plasma lipoproteins and of albumin proceed by the same mechanism, which involves a steric hindrance for these macromolecules in the influx pathway.

動脈壁内のいわゆる“クリアランス”について

The insudation of blood plasma components into the intima is one of the most important atherogenic factors. But, the accumulation of blood components would not occur when the intimal fluid transportation (“clearance”) is satisfactory enough to carry them away.We examine microscopically the declining of intimal clearance with aging about non-sclerotic areas of human aortic wall by Berlin-blue production after infusion of ferric citrate solution.The infusion of fluid was much easier and more uniform in the younger aortas than in older cases, and it was washed away by perfusing of saline solution. But in older cases, blue dye remained distinctly in the intima.It was noted that the clearance mechanism of human aortic wall declined steadily with aging even in the non-sclerotic “normal” areas.

Transfer of plasma lipoprotein components and of plasma proteins into aortas of cholesterol-fed rabbits. Molecular size as a determinant of plasma lipoprotein influx.

The arterial influx of esterified and free cholesterol from low density lipoproteins and very low density lipoproteins in 20 hypercholesterolemic rabbits was measured simultaneously by the use of lipoproteins labeled in vivo with [3H]- and [14C]-cholesterol. The simultaneous arterial influx of either [3H]-leucine-labeled very low density lipoproteins, low density lipoproteins, high density lipoproteins, or plasma proteins was also measured in each rabbit. The arterial influx was calculated as intimal clearance, i.e., the influx of a given fraction divided by its plasma concentration. The intimal clearance of low density lipoprotein esterified cholesterol was equal to that for the apolipoproteins of that fraction, which is compatible with an arterial influx of intact low density lipoprotein molecules. The intimal clearance of very low density apolipoprotein or cholesteryl ester was less than that for low density lipoprotein, whereas high density lipoprotein and albumin clearances exceeded low density lipoprotein clearance by 1.5- to 3-fold. The intimal clearances of plasma proteins, high density, low density, and very low density lipoproteins decreased linearly with the logarithm of the macromolecular diameter. This indicates that the arterial influx of three plasma lipoprotein fractions and of plasma proteins proceeds by similar mechanisms. Apparently the relative intimal clearances of lipoproteins are more dependent on their size relative to pores or vesicular diameters at the plasma-artery interface than on specific interactions between lipoproteins and the arterial intimal surface.