[show abstract][hide abstract] ABSTRACT: Diabetic patients are at high risk of cardiovascular disease and the risk is amplified in the presence of nephropathy, which may be partially attributed to modifications in lipoproteins. Moreover, lipoprotein profile may be affected by incipient nephropathy, glomerulopathy, and mild or severe renal failure. The aim of our study was to evaluate whether chronic renal failure (CRF) changes lipoprotein profile and apo A-I urinary excretion in diabetic subjects with glomerulopathy in comparison with non-diabetic subjects with glomerulopathy and CRF. Diabetic (n=25) and non-diabetic (n=10) patients with glomerulopathy and CRF showed significantly higher LDL-cholesterol, non-HDL-cholesterol and HDL-triglyceride levels than diabetic individuals without CRF (n=10). Arylesterase and paraoxonase activities did not show any difference between groups. Apo A-I could not be detected in urine samples from diabetic patients without CRF. All diabetic subjects with glomerulopathy and CRF who presented proteinuria above 6.5 g/24 h showed detectable urinary apo A-I (range=13.1-61.0 mg/24 h). Similarly, all non-diabetic patients with glomerulopathy and CRF who had proteinuria above 8.0 g/24 h also evidenced detectable apo A-I in urine (range=25.6-557.3 mg/24 h). Urinary apo A-I showed positive and significant correlations with urea (r=0.73, p<0.05) and proteinuria (r=0.97, p<0.0001), and a negative correlation with albumin plasma levels (r=-0.68, p<0.05). In conclusion, the presence of CRF in diabetic patients was associated with a more atherogenic lipoprotein profile.
Diabetes Research and Clinical Practice 02/2007; 75(1):35-41. · 2.74 Impact Factor
[show abstract][hide abstract] ABSTRACT: Physical activity is known to play a cardioprotective role. Nevertheless, a paradox seems to arise when considering that aerobic exercise enhances oxidative stress. In previous works, we showed that free radical formation during physical activity was counteracted by an increase in antioxidant defenses. Low density lipoprotein (LDL) oxidation is a crucial step in atherosclerosis, process that can be inhibited by high density lipoprotein (HDL) through its oxidable components or associated enzymes like paraoxonase (PON) and platelet-activating factor acetylhydrolase (PAF-AH). In this study, we evaluated copper-induced oxidation in isolated LDL and HDL fractions, and the effect of HDL on LDL oxidation in samples from well trained amateur athletes who were participating in an ultra-distance triathlon (n=18) in comparison with healthy sedentary controls (n=18). PON and PAF-AH activities and PON phenotype were also evaluated. The oxidability of isolated lipoproteins, as well as HDL antioxidant capacity, was similar in both groups of subjects. After classification by paraoxonase phenotype, only sportsmen belonging to the QR phenotype showed higher HDL susceptibility to in vitro oxidation (thiobarbituric reactive substances, TBARS) than controls (p<0.05). HDL oxidability exhibited a positive correlation with its triglyceride content (r=0.58; p<0.01). Similarly, HDL capacity to inhibit LDL oxidation was increased in athletes (p<0.05) which was positively associated with HDL oxidability (HDL-TBARS: r=0.55, p<0.005; HDL-lag time: r=0.45, p<0.01; HDL-D max: r=0.35, p<0.05). In conclusion, regular aerobic exercise was associated to a more efficient antioxidant function played by HDL from PON-QR carriers, which could constitute an adaptive response to the increased oxidative stress.
Life Sciences 05/2006; 78(26):3074-81. · 2.56 Impact Factor
[show abstract][hide abstract] ABSTRACT: The high incidence of atherosclerosis in women after menopause is associated with a risk pattern including an increase in low density lipoprotein (LDL), even though high density lipoprotein (HDL) cholesterol levels tend to be maintained or slightly decreased. Since estrogens are considered potent antioxidants, an increase in lipid peroxidation and formation of reactive oxygen species would be expected after menopause. If HDL becomes oxidized, the ability to protect LDL against oxidation may be impaired. In postmenopausal women there are scarce reports concerning HDL oxidability and no data about its antioxidant activity. We studied copper-induced oxidation and conjugated dienes formation in HDL isolated from 58 women, 30 postmenopausal (PMW) and 28 premenopausal (PreMW). None presented diabetes or cardiovascular disease and none was receiving hormonal, hypolipidemic or antioxidant therapy either. In order to evaluate the effect of HDL on LDL oxidation we isolated LDL and HDL from the same subject and assessed copper-induced LDL oxidation in the presence of HDL, followed by thiobarbituric acid-reactive substances determination. Relationships with HDL chemical composition, alpha-tocopherol content, cholesteryl ester transfer protein (CETP) and paraoxonase activity (PON) were investigated. HDL chemical composition in PMW exhibited triglyceride enrichment when compared to PreMW (p <0.05). alpha-Tocopherol content and CETP activity were similar in both groups. However, CETP activity correlated positively with HDL triglyceride and negatively with HDL cholesterol percentage (r=0.44, p <0.01 and r=-0.32, p <0.05, respectively). Paraoxonase activity did not show differences between PMW and PreMW. When evaluating HDL oxidability, PMW revealed a shorter lag time in comparison to PreMW, even after adjustment for age, p <0.05. Moreover, when the effect of HDL on LDL oxidation was evaluated, HDL from PMW showed a reduction in its ability to inhibit LDL oxidation, compared to PreMW (p <0.05). In addition, the extent of inhibition of LDL oxidation by HDL was positively correlated with HDL resistance to oxidation (r=0.27, p <0.05). After women classification by paraoxonase phenotype, HDL ability to protect LDL against oxidation remained reduced only in PMW belonging to the PON QR phenotype, in comparison to PreMW QR. These results suggest that HDL from PMW exhibits impairment in its antioxidant ability, which is associated to a decreased HDL resistance to oxidation. In turn, this was related to triglyceride enrichment of HDL particles. All these alterations were independent from HDL cholesterol plasma levels.
[show abstract][hide abstract] ABSTRACT: It is widely accepted that aerobic physical activity is associated with a less atherogenic lipid and lipoprotein profile and, consequently, with a reduced cardiovascular risk. Both cross-sectional studies and prospective-interventional trials show that the most frequent modification observed consists of a slight but significant increase in high-density lipoprotein cholesterol (HDL-C) levels. Nevertheless, only few studies made an attempt to elucidate if this quantitative modification was accompanied by an improvement in any of HDL antiatherogenic functions. The purpose of this study was to evaluate the main steps of reverse cholesterol transport, the best known antiatherogenic function performed by HDL, in a group of well-trained soccer players (n = 35) in comparison to sedentary controls (n = 15). Average HDL-C levels were 12.5% higher in the sportsmen, in large part because of greater HDL2-C concentration. No statistically significant differences were observed in the other lipid- and lipoprotein-related parameters. The capacity to promote cholesterol efflux from Fu5AH cells was significantly higher in the soccer players than in the control individuals (20.5% +/- 0.4% v 15.9% +/- 1.2%, respectively, P < .001). However, lecithin:cholesterol acyltransferase (LCAT; 2.6 +/- 0.9 v 1.4 +/- 0.3%/mL.h, respectively) and cholesteryl ester transfer protein (CETP; 69.5 +/- 8.3 v 62.7 +/- 14.8%/mL.h, respectively) activities did not reach statistically significant difference between both groups. Correlation analysis showed that cholesterol efflux induced by serum samples was directly related to HDL-C (r = 0.59, P < .001), HDL2-C (r = 0.37, P < .01), and lipoprotein (Lp)A-I (r = 0.44, P < .05). On the other hand, negative correlations were observed with waist/hip ratio (r = -0.36, P < .05), low-density lipoprotein cholesterol (LDL-C; r = -0.33, P < .05), apolipoprotein B (apo B; r = -0.42, P < .05), and LpA-I;A-II (r = -0.51, P < .005). In conclusion, the well-known cardioprotective benefit of regular exercise could be based, at least in part, on a less atherogenic lipid and lipoprotein profile and an enhanced cellular cholesterol efflux.
[show abstract][hide abstract] ABSTRACT: We have generated transgenic rabbits that express the entire human apoA-I/C-III/A-IV gene cluster. As in humans, h-apoA-I and h-apoC-III were expressed in liver and intestine, whereas h-apoA-IV mRNA was detected in intestine only. Transgenic rabbits had significantly higher plasma total cholesterol, HDL-cholesterol and total phospholipid concentrations than non-transgenic littermates. In contrast to similar transgenic mice previously generated, which have gross hypertriglyceridemia, triglyceride concentrations were only moderately raised in transgenic rabbits. Plasma and HDL from transgenic rabbits were more effective than those from controls in promoting cholesterol efflux from cultured hepatoma cells. They had lower LCAT, lower CETP and higher PLTP activities than non-transgenic littermates. Cholesterol-feeding produced major increases in plasma lipids. The qualitative response to the diet was not modified by cluster expression. Human apoA-I concentration was halved by cholesterol-feeding, whereas h-apoC-III and h-apoA-IV concentrations were not significantly altered. Cholesterol efflux from hepatoma cells to plasma and HDL was not altered by the diet. Since lipoprotein metabolism of rabbits closely resembles that of humans, human apoA-I/C-III/A-IV transgenic rabbits may provide a reliable model for studies of the transcriptional regulation of the cluster, and for evaluating the effects of different agents on the expression of the three genes.
[show abstract][hide abstract] ABSTRACT: Lipoprotein lipase (LPL) deficiency is a rare autosomal recessively inherited disease characterized by elevated triglyceride, low total cholesterol and quantitative and qualitative alterations of high-density lipoprotein (HDL). The aim of the present study was to explore HDL metabolic activities in a patient with LPL deficiency and in his family (n = 11).
Subjects were divided into four groups: proband (Ser447Stop/Arg170Leu carrier), Ser447Stop carriers, Arg170Leu carriers and silent mutation/wild-type carriers (controls). Cholesterol efflux from Fu5AH cells, lecithin:cholesterol acyltransferase (LCAT), cholesteryl ester transfer protein (CETP), paraoxonase 1 (PON1) and platelet-activating factor acetylhydrolase (PAF-AH) activities were evaluated.
Comparison between the proband and the control group revealed that the boy had significantly reduced cholesterol efflux (P < 0.001), conserved LCAT activity (P > 0.05) and increased CETP activity (P < 0.001). As regards antioxidant enzymes, while PON1 activity was higher in the proband than in the controls (P < 0.0001), PAF-AH activity was reduced (P < 0.05). The other groups did not show relevant differences in comparison with controls.
The presence of one mutation was not enough to introduce important modifications in HDL functions. Markedly reduced HDL levels can keep certain normal enzymatic activities, which probably tend to counteract the deleterious effects of LPL deficiency.
European Journal of Clinical Investigation 07/2004; 34(7):467-74. · 3.37 Impact Factor
[show abstract][hide abstract] ABSTRACT: Previous studies have shown that high density lipoprotein (HDL)-deficient states are associated with reduced paraoxonase 1 (PON1) activity. However, HDL reduction caused by primary hypertriglyceridemia has not been fully explored. The aim of the present study was to evaluate whether PON1 and platelet-activating factor acetylhydrolase (PAF-AH), two antioxidant enzymes, were altered in patients with low HDL-cholesterol levels with or without primary hypertriglyceridemia in comparison with control normolipemic subjects.
We studied 24 patients with low HDL-cholesterol levels with (n=12) or without (n=12) primary hypertriglyceridemia in comparison with 12 control subjects who presented normal HDL-cholesterol and triglyceride levels. Paraoxon and phenylacetate were used as substrate for measuring PON1 activities and 1-hexadecyl-2-[3H]acetyl-glycero-3-phosphocholine for platelet-activating factor acetylhydrolase (PAF-AH) activity. Double substrate method was used to assign phenotypes. Lipid, lipoprotein, apolipoprotein, and lipoprotein particles were determined by standardized methods.
Both PON1 activities were significantly reduced in patients with low HDL-cholesterol levels. This reduction could be selectively attributed to the hypertriglyceridemic subgroup. PAF-AH activity was not different between hypoalphalipoproteinemic patients and controls. PON1 activities correlated positively and significantly with HDL-cholesterol, HDL2-cholesterol, HDL3-cholesterol, HDL-phospholipids, apo A-I, apo A-II, and LpA-I:A-II. PAF-AH correlated positively and significantly with total and low density lipoprotein-cholesterol.
Data from this study would suggest that in hypoalphalipoproteinemic syndrome, particularly when associated with hypertriglyceridemia, there is impairment in enzymatic antioxidant activity exclusively related with HDL.
Archives of Medical Research 01/2004; 35(3):235-40. · 2.08 Impact Factor
[show abstract][hide abstract] ABSTRACT: Even if physical activity constitutes a well-known antiatherogenic factor, the precise mechanisms underlying this protective effect are not completely clear.
Lipid and antioxidant profiles were evaluated in 15 well-trained rugby players and 15 sedentary controls. Lipoprotein fractions were separated by sequential ultracentrifugation and alpha-tocopherol content was determined in each fraction by high-performance liquid chromatography. Susceptibility to in vitro oxidation was also measured in intermediate and low density lipoproteins isolated from both groups of subjects as the production of conjugated dienes.
Although the sportsmen were not receiving any special diet or vitamin supplementation they showed a slightly improved lipoprotein profile, mainly represented by increased high density lipoprotein-cholesterol levels (P < 0.05), and an enhanced antioxidant status. The latter was evidenced by an increment in total radical antioxidant potential (P < 0.001), higher ascorbic acid (P < 0.005) and alpha-tocopherol (P < 0.05) plasma concentrations, and elevated activities of superoxide dismutase (P < 0.001) and arylesterase (P < 0.01). Moreover, only the fraction of intermediate and low density lipoproteins from rugby players presented higher alpha-tocopherol content in comparison with sedentary controls (484 +/- 67 vs. 377 +/- 123 microg dL(-1), respectively; P < 0.01). Nevertheless, the susceptibility to in vitro oxidation of this lipoprotein fraction was not different between both groups.
Given that intermediate density and low density lipoproteins represent the most atherogenic fraction, this finding, in combination with the improved lipid and antioxidant status, would add to the link between regular physical activity and protection against cardiovascular disease.
European Journal of Clinical Investigation 12/2002; 32(11):818-25. · 3.37 Impact Factor
[show abstract][hide abstract] ABSTRACT: Low density lipoprotein (LDL) oxidation is a crucial step in the atherosclerotic process. High density lipoprotein (HDL)-associated enzymes such as paraoxonase could exert a protective effect on LDL oxidation in the arterial wall, an effect which could be impaired in Type 2 diabetes mellitus (T2DM). We studied copper-induced oxidation in LDL and HDL isolated from 17 T2DM patients with fair glycaemic control and HDL-cholesterol within normal range and 17 healthy normolipidaemic control subjects. To evaluate the effect of HDL on LDL oxidation in diabetic and control subjects, we assessed copper-induced oxidation in HDL/LDL mixtures, with each lipoprotein isolated from the same subject. Relationships with HDL chemical composition, alpha-tocopherol content and serum paraoxonase activity were investigated. Oxidation was promoted by lipoprotein incubation with copper and then thiobarbituric acid reactive substances (TBARS), conjugated diene production and electrophoretic mobility in agarose gel were measured. In T2DM subjects HDL oxidation was higher than in controls. However, HDL from diabetics was as effective as control HDL to inhibit LDL oxidation. Neither HDL chemical composition nor serum paraoxonase activity showed any difference as compared to control subjects. In contrast, HDL from T2DM subjects showed a higher alpha-tocopherol content which positively correlated with HDL oxidability. Paraoxonase activity positively and strongly correlated with HDL inhibitory effect on LDL oxidation in patients and controls belonging to the heterozygous activity phenotype. Besides, LDL oxidability showed no differences between patients and controls. These results suggest that fairly-controlled T2DM patients with HDL-cholesterol levels within normal range show: 1) normal HDL ability to inhibit LDL oxidation related to normal paraoxonase activity; 2) higher HDL oxidability in spite of its high alpha-tocopherol content, which could favour tocopherol-mediated peroxidation and 3) normal LDL oxidability possibly due to the lack of significant lipoprotein structural alterations.