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The Journal of Clinical Endocrinology & Metabolism Vol. 89, No. 9 4721-4728
Copyright © 2004 by The Endocrine Society

The Influence of Lipoprotein Lipase Gene Variation on Postprandial Lipoprotein Metabolism

José López-Miranda, Gracia Cruz, Purificación Gómez, Carmen MarÍn, Elier Paz, Pablo Pérez-MartÍnez, Francisco J. Fuentes, José M. Ordovas and Francisco Pérez-Jiménez

Lipids and Atherosclerosis Research Unit (J.L.-M., G.C., P.G., C.M., E.P., P.P.-M., F.J.F., F.P.-J.), Reina Sofía University Hospital, 14004 Córdoba. Spain; and Nutrition and Genomics Laboratory (J.M.O.), Tufts University, Boston, Massachusetts 02111

Address all correspondence and requests for reprints to: Francisco Pérez Jiménez, Unidad de Lípidos y Arteriosclerosis, Hospital Universitario Reina Sofía, Avda Menéndez Pidal, s/n. 14004 Córdoba, Spain. E-mail: md1pejif{at}uco.es.


   Abstract
Top
 Abstract
Introduction
 Subjects and Methods
 Results
 Discussion
 References
 
Lipoprotein lipase (LPL) is one of the key enzymes in the metabolism of triacylglycerol-rich lipoproteins (TRL). We evaluated whether the association of LPL HindIII (H1/H2) and Serine447-Stop (S447X) polymorphisms may explain the interindividual variability observed during postprandial lipemia. Fifty-one healthy male volunteers (26 with the H2S447 genotype, 15 with the H1X447 genotype, and 10 with the H1S447 genotype) were subjected to a vitamin A-fat load test consisting of 1 g fat/kg body weight and 60,000 IU vitamin A. Blood was drawn every hour until the 6th hour and every 2 h and 30 min until the 11th hour. Data revealed that subjects that are homozygous for the H2 allele (H2H2) showed a higher postprandial response for small TRL, retinyl palmitate (RP), large TRL-RP, large TRL-B48, and small TRL-B48 levels. Furthermore, in the case of the S447X polymorphism, 447Ter carriers had a lower postprandial response for small TRL-RP, large TRL-B48, and small TRL-RP. Subjects with the LPL H2S447 genotype had higher plasma triacylglycerol, large TRL-triacylglycerol, large TRL-RP, small TRL-RP, and large TRL-B48 (P < 0.037) than H1X447 subjects. The modifications observed in postprandial lipoprotein metabolism in young normolipemic males with LPL polymorphism could be involved in the lower risk of coronary artery disease associated with the H1X447 genotype.


   Introduction
Top
 Abstract
 Introduction
Subjects and Methods
 Results
 Discussion
 References
 
LIPOPROTEIN LIPASE (LPL) is an enzyme that is located on the luminal surface of vascular endothelium. Its best known function consists of hydrolyzing triacylglycerols (TG) in the chylomicrons and very-low-density lipoproteins (VLDL), thus initiating the lipolytic cascade required to convert VLDL into low-density lipoproteins (LDL) (1, 2). It also favors hepatic uptake of lipoprotein remnants (3) and can act as a ligand in the binding of LDL to the extracellular matrix (4, 5) and in its uptake by macrophages (6).

The presence of two defective alleles in the gene locus of LPL causes a severe deficit in the activity of this enzyme, producing pronounced fasting hypertriacylglycerolemia (7, 8). Other genetic variations, which are relatively common in the general population, can produce partial LPL deficit and are associated with changes in the lipid profile and increased coronary risk (9, 10). The polymorphism determined by the HindIII restriction enzyme consists of replacing thymidine with guanine at position 495 of intron 8 of this gene (11). This change deletes the recognition sequence of the HindIII enzyme and permits allele H1 to be distinguished from allele H2. The H2H2 genotype has been associated with elevated fasting TG (12, 13) and LDL cholesterol (LDL-C) levels (13), a drop in high-density lipoprotein (HDL) cholesterol (HDL-C) (14), primary hypertriacylglycerolemia (15), and premature coronary artery disease (CAD) (12, 16). Another frequent polymorphism has been identified that alters the penultimate amino acid Serine447 to a stop codon (S447X) resulting in a truncation of the enzyme (17) and higher expression in vitro studies (18). Although the results of early studies that examined the association between the S447X polymorphism and lipid levels were inconsistent (19), the X447 allele occurs less frequently in patients with hypertriacylglycerolemia than in healthy whites and is more common in healthy control subjects than in myocardial infarction patients (20). Recent studies have clearly demonstrated that X447 mutation is associated with higher postheparin LPL activity in patients (21) and a favorable lipid profile, with lower plasma cholesterol, lower TG, and higher HDL-C levels (12, 22). Thus, there is strong evidence to suggest that the H1X447 genotype is associated with a beneficial lipid profile and that it may therefore offer protection against CAD (23). Furthermore, subjects in this study with the H1X447 genotype had a lower postprandial response in plasma TG than the H2 subjects. Triglyceride-rich lipoproteins are a heterogeneous population of particles that differ in origin, structure, and cell receptor interactions and have different physiological significance. However, the lipoprotein fractions involved were not determined in the latter study.

Since 1979, when Zilversmit (24) proposed that triacylglycerol-rich lipoproteins (TRL) played a role in arteriosclerosis, many research teams have shown the role of postprandial lipoprotein particles in the development of CAD (25, 26, 27, 28). Because LPL is the rate-limiting enzyme for the hydrolysis of TG, LPL activity is critical for normal clearance of postprandial triglyceride-rich lipoprotein particles. Thus, the aim of this study was to determine whether the HindIII and S447X LPL polymorphisms could modify the postprandial response of TRL in young normolipemic males to explain the lower risk of CAD associated with the H1X447 genotype.


   Subjects and Methods
Top
 Abstract
 Introduction
 Subjects and Methods
Results
 Discussion
 References
 
Human subjects

Initially, one hundred fifteen male subjects, who were students at the University of Cordoba, volunteered to participate in the study in response to an advertisement. They ranged from 18–49 yr of age. All underwent a comprehensive medical history, physical examination, and clinical chemistry analysis before enrollment. The group underwent phenotype testing for the apolipoprotein (Apo) E, with the object of selecting those presenting the phenotype most widespread in the general population (E3E3), thus eliminating the effect triggered by this genetic variation on both postprandial lipemic response and on plasma lipid levels. Finally, fifty-one healthy male subjects with the Apo E3/3 agreed to take part in the study. Twenty-five were carriers of the H1 allele in both its homozygous and heterozygous form, and 26 were carriers of the H2H2 genotype for the HindIII polymorphism. Furthermore, 15 were carriers of the 447Ter allele in both its homozygous and heterozygous form, and 36 were homozygotes for the 347Ser allele for the LPL447 mutation. Twenty-six subjects with the H2S447 genotype, 15 with the H1X447 genotype, and 10 with the H1S447 genotype were selected after genotype analysis. Because of complete linkage disequilibrium between HindIII and S447, we have detected only three genotypes: H2S447, H1S447, and H1X447 (with the exception of one recombinant). None of them had liver, renal, or thyroid disease or diabetes. Each of the subjects selected had the Apo E3/3 genotype, to avoid allele effects of this gene locus on postprandial lipemia (29). They were not taking medication or vitamins known to affect plasma lipids. The fasting plasma lipid, lipoprotein, Apo levels, age, and body mass index (BMI) according to LPL polymorphism are shown in Table 1Go. All studies were carried out in the Research Unit of the Reina Sofia University Hospital. The Human Investigation Review Committee approved the experimental protocol at the Reina Sofia University Hospital.


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  		TABLE 1. Characteristics of the subjects at baseline according to LPL polymorphisms

 
Vitamin A fat-loading test

After a 12-h fast, subjects were given a fatty meal enriched with 60,000 U vitamin A/m2 body surface area. The fatty meal consisted of two cups of whole milk, eggs, bread, bacon, cream, walnuts, and butter, which was consumed within 20 min. The meal provided 1 g fat and 7 mg cholesterol/kg body weight and contained 60% fat, 15% protein, and 25% carbohydrates. After the meal, subjects were not allowed to consume any calorie-containing food for 11 h. Blood samples were drawn before the meal, every hour until the 6th hour, and every 2nd hour and 30 min until the 11th hour.

Lipoprotein separations

Blood was collected in tubes containing EDTA to give a final concentration of 0.1% EDTA. Plasma was separated from red cells by centrifugation at 1,500 x g for 15 min at 4 C. The chylomicron and large VLDL fraction of TRL was isolated from 4 ml plasma overlayered with 0.15 M NaCl, 1 mM EDTA (pH 7.4; density, 1.006 g/ml) by a single ultracentrifugal spin (28,000 x g, 30 min, 4 C) in a 50-type rotor (Beckman Instruments, Fullerton, CA). Chylomicrons contained in the top layer were removed by aspiration after cutting the tubes. The infranatant was centrifuged at a density of 1.019 g/ml for 24 h at 115,000 x g in the same rotor. The nonchylomicron fraction of TRL (also referred to as small TRL) was removed from the top of the tube. All operations were done in subdued light. Large and small TRL fractions were stored at –70 C until assayed for retinyl palmitate (RP).

Lipid analysis

Cholesterol and TG in plasma and lipoprotein fractions were assayed by enzymatic procedures (30, 31). Apo A-I and Apo B were determined by turbidimetry (32). HDL-C was quantified by analyzing the supernatant obtained after precipitation of a plasma aliquot with dextran sulfate-Mg2+, as described by Warnick et al. (33). LDL-C was calculated as the difference between the cholesterol at the bottom of the tube after ultracentrifugation at 1.019 g/ml and the HDL-C.

RP assay

The RP content of large and small TRL fractions was assayed using a method previously described (34). Briefly, different volumes of the various fractions (100 µl for chylomicrons and 100–500 µl for remnants) were placed in 13 x 100 mm glass tubes. The total volume in each tube was adjusted, as needed, to 500 µl with use of isotonic sodium chloride solution. Retinyl acetate (40 ng in 200 µl mobile phase buffer) was added to each tube as an internal standard. Five hundred milliliters of methanol were added, followed by 500 µl of the mobile phase buffer, for a total vol of 1.7 ml. The mobile phase buffer was prepared fresh on a daily basis by combining 90 ml hexane, 15 ml n-butyl chloride, 5 ml acetonitrile, and 0.01 ml acetic acid (82:13:5 by volume with 0.01 ml of acetic acid). The tubes were thoroughly mixed after each step. The final mixture was centrifuged at 350 x g for 15 min at room temperature, and the upper layer was carefully removed by aspiration and placed into individual autosampler vials. The autoinjector was programmed to deliver 100 µl/injection and a new sample every 10 min in a custom prepackaged silica column SupelcoSil LC-SI (5 mm, 25 cm x 4.6 mm inner diameter) provided by Supelco Inc. (Bellefonte, PA). The flow was maintained at a constant rate of 2 ml/min, and the peaks were detected at 330 nm. The peaks of RP and retinyl acetate were identified by comparing retention time with a purified standard (Sigma, St. Louis, MO). The RP concentration in each sample was expressed as the ratio of the area under the RP peak to the area under the retinyl acetate peak (35). All operations were performed in subdued light.

Determination of ApoB-48 and ApoB-100

ApoB-48 and ApoB-100 were determined by SDS-PAGE as described by Karpe and Hamsten (36). Electrophoretic separation was performed using a 3–20% gradient polyacrylamide gel with a vertical Hoeffer Mighty Small II electrophoresis apparatus (Hoeffer Pharmacia Biotech, San Francisco, CA). Gels were scanned with a video densitometer scanner (TDI, Madrid, Spain) connected to a personal computer to integrate signals. Background intensity was calculated after scanning an empty lane. The coefficient of variation for the SDS-PAGE was 7.3% for ApoB-48 and 5.1% for ApoB-100.

DNA amplification and genotyping of Apo E, Apo AIV, and LPL polymorphism

DNA was extracted from 10 ml EDTA-containing blood. Amplification of a region of 266-bp of the Apo E gene was performed by PCR with 250 ng genomic DNA and 0.2 µmol of each oligonucleotide primer (E1, 5'-GACACTGACCCCGGTGGCGGAG-3', and E2, 5'-TCGCGGGCCCCGGCCTGGCCTGGTACACTGCCA-3'), and 10% dimethylsulfoxide in 50 µl DNA was denatured at 95 C for 5 min followed by 30 denaturation cycles at 95 C for 1 min, annealing at 63 C for 1.5 min, and extension at 72 C for 2 min. Twenty microliters of the PCR product were digested with 10 U restriction enzyme CfoI (BRL, Gaithersburg, MD) in a total volume of 35 µl. Digested DNA was separated by electrophoresis on an 8% nondenaturing polyacrylamide gel at 150 V for 2 h. Bands were visualized by silver staining.

Identification of the genotypes for the Apo AIV polymorphism was done by amplification of a fragment of gene. A total of 0.5 µg genomic DNA and 1.25 U Taq polymerase were used with 200 µmol of nucleotides and 0.3 µmol of each primer oligonucleotide (5'-GCCCTGGTGCAGCAGATGGAACAGCTCAGG-3' y 5'-CATCTGCACCTGCTCCTGCTGCTGCTCCAG-3') in a final vol of 50 µl. DNA was denatured at 95 C for 5 min followed by 30 denaturation cycles at 95 C for 1 min, annealing at 55 C for 1.5 min, and extension at 72 C for 2 min. Twenty microliters of the PCR product were digested with 10 U HindIII restriction enzyme (BRL) in a total vol of 35 µl. The S447X polymorphism in exon 9 was identified by the introduction of a forced HinfI restriction enzyme site into the PCR product. Digested DNA was separated by electrophoresis on an 8% nondenaturing polyacrylamide gel at 150 V for 2 h. Bands were visualized by silver staining.

Identification of the genotypes for the LPL HindIII polymorphism was done by amplification of a fragment of 365 bp in intron 8 of the gene. A total of 0.5 µg genomic DNA and 1.25 U of Taq polymerase were used with 200 µmol nucleotides and 0.3 µmol of each primer oligonucleotide (5'-TTTAGGCCTGAGTTTCCAC-3' y 5'-CTCCCTAGACAGAGATC-3') in a final vol of 50 µl. DNA was denatured at 95 C for 5 min followed by 30 denaturation cycles at 95 C for 1 min, annealing at 55 C for 1.5 min, and extension at 72 C for 2 min. Twenty microliters of the PCR product were digested with 10 U HindIII restriction enzyme (BRL) in a total vol of 35 µl. The S447X polymorphism in exon 9 was identified by the introduction of a forced HinfI restriction enzyme site into the PCR product. The following primers were used: forward primer 5'-CATCCATTTTCTTCCACGGG-3' and reverse primer 5'-TAGCCCAGAATGCTCACCAGACT-3'. A total of 0.5 µg genomic DNA and 1.25 U Taq polymerase were used with 200 µmol nucleotides and 0.3 µmol of each primer oligonucleotide in a final vol of 50 µl. DNA was denatured at 95 C for 5 min followed by 30 denaturation cycles at 94 C for 1 min, annealing at 55 C for 1 min, and extension at 72 C for 1 min. Twenty microliters of the PCR product were digested with 10 U HinfI restriction enzyme (BRL) in a total vol of 35 µl. Digested DNA was separated by electrophoresis on an 8% nondenaturing polyacrylamide gel at 150 V for 2 h. Bands were visualized by silver staining.

Statistical analysis

The following variables were calculated to characterize the postprandial responses of plasma TG, large TRL, and small TRL to the test meal: 1) area under the curve (AUC), which was the area between the plasma concentration vs. time curve and baseline drawn parallel to the horizontal axis through the 0 h concentration (a computer program using the trapezoidal rule calculated this area); 2) peak concentration, which was the average of the peak and the second highest concentration above the baseline; and 3) the peak time, which was the average time to peak concentration and the time to the second highest concentration. Data were tested for statistical significance between genotypes and time by ANOVA for repeated measures. In this analysis, we studied the statistical effects of the genotype (represented as G) independent of the time in the postprandial study, the effect of time alone, or change in the variable after ingesting fatty food over the entire lipemic period (represented as T) and the effect of the interaction of both factors, genotype and time, which is indicative of the magnitude of the postprandial response in each group of subjects with a different genotype (represented as GxT). The means were adjusted for BMI and age. When statistical significance was found, Tukey’s post hoc comparison test was used to identify group differences. Because of complete linkage disequilibrium between HindIII and S447 polymorphism, only three genotypes existed: H2S447, H1S447, and H1X447. Probability values under 0.05 were considered significant. Stepwise multiple regression analyses were carried out using small and large TRL-RP normalized peak concentration and AUC as dependent variables and age, BMI, LPL genotypes, basal cholesterol, TG, and HDL-C values as independent variables. Discrete variables were classified for analysis. All data presented in the text and tables are mean values ± SD.


   Results
Top
 Abstract
 Introduction
 Subjects and Methods
 Results
Discussion
 References
 
The baseline characteristics for the combined genotypes are shown in Table 1Go. No differences were observed in any of the lipid parameters studied, age, or BMI between the three groups of subjects studied. Because of complete linkage disequilibrium between the HindIII and S447 polymorphism, only three genotypes existed: H2S447 (n = 26), H1S447 (n = 10), and H1X447 (n = 15).

We first performed an analysis of the independent effect of each polymorphism of the LPL (HindIII and S447X polymorphisms) on postprandial lipemic response. Homozygotes for the H2 allele (H2H2) showed a significantly higher postprandial response for triglyceride-rich lipoproteins (TRL) of intestinal origin than subjects with the H1 allele, as they had a higher postprandial response for levels of RP detected in both triglyceride-rich fractions. The AUC for RP and Apo B48 in large TRL and small TRL was greater in H2H2 subjects than H1 subjects (Table 2Go). In the case of the S447X polymorphism, carriers of the 447Ter allele had a lower postprandial response for RP in chylomicron remnants and for Apo B48 in both large and small TRL. Thus, the AUC was greater in homozygotes for the 447Ser allele than in carriers of the 447Ter allele (Table 3Go).


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  		TABLE 2. AUC for the different lipid parameters examined according to HindIII polymorphism

 

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  		TABLE 3. AUC for the different lipid parameters examined according to Ser447-Ter of LPL polymorphism

 
We then performed an analysis by genotypes combining both genotypes. Plasma TG, large TRL-TG, and small TRL-TG responses after the fat load test are shown in Fig. 1Go. Significant time effects were found for plasma TG (P < 0.001), large TRL-TG (P < 0.001), and small TRL-TG (P < 0.001), demonstrating that the levels of TG increase during the postprandial period in plasma and both TRL fractions. There were no significant differences in small TRL-TG response among the three subject groups as shown by the ANOVA in which G indicates the effect of genotype, T the effect of time, and GxT the effect of the interaction genotype-time. Carriers of the LPL H2S447 genotype had the highest plasma TG levels (Fig. 1AGo) (P < 0.045) and large TRL-TG (Fig. 1BGo) (P < 0.05) postprandial response. An intermediate response was found in subjects with the H1S447 genotype; whereas the lowest postprandial response was observed in subjects with the H1X447 genotype.



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  		FIG. 1. Line plots of postprandial plasma triglyceride (A), large TRL-triglyceride (B), and small TRL-triglyceride (C) in H1X447 (dotted line, •), H1S447 (solid line, {circ}), and H2S447 subjects (dotted line, {blacktriangleup}) (H1 indicates H1H1 or H1H2 genotypes). For each group, the levels at each time point were averaged. Systeme International conversion factors are: 0.02586 mmol/liter for cholesterol and 0.01129 mmol/liter for TG. G, Genotype effect; T, time effect; GxT, genotype by time interaction. ANOVA for repeated measures.

 
Plasma levels of RP in the large and small TRL showed a significant increase above basal levels in both genotypes during the postprandial period (Fig. 2Go, B and C). In this case, the LPL genotype also had a significant effect on H2S447 subjects, presenting a greater postprandial response of RP in the large TRL (P < 0.009) and small TRL (P < 0.013) than subjects with the H1X447 genotype. In addition, the AUC of large TRL-RP (P < 0.004) and small TRL-RP (P < 0.014) was greater in H2S447 subjects than in carriers of the H1X447 genotype (Table 4Go). In addition, the AUC of large TRL ApoB-48 was higher in H2S447 subjects (P < 0.036) than in carriers of the H1 allele (Fig. 2AGo and Table 4Go). There were no significant differences in the AUC of the remaining lipid parameters in either group of subjects (Table 4Go).



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  		FIG. 2. Line plots of postprandial large TRL-Apo B48 (A), large TRL-RP (B), and small TRL-RP (C) in H1X447 (dotted line, •), H1S447 (solid line, {circ}), and H2S447 subjects (dotted line, {blacktriangleup}) (H1 indicates H1H1 or H1H2 genotypes). For each group, the levels at each time point were averaged. Systeme International conversion factors are: 0.02586 mmol/liter for cholesterol and 0.01129 mmol/liter for TG. G, Genotype effect; T, time effect; GxT, genotype by time interaction; AU, arbitrary units. ANOVA for repeated measures.

 

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  		TABLE 4. AUC for the different lipid parameters examined according to LPL polymorphism adjusted by BMI and age as covariant

 
Multiple regression analysis (Table 5Go) revealed that the LPL polymorphism independently predicts the variability of postprandial response of the large TRL-RP AUC (P < 0.001) and the variability of the small TRL-RP (P < 0.007) in this population group. Age, BMI, TG, and Apo AIV polymorphism had no effect on the response.


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  		TABLE 5. Multiple stepwise regression analyses

 

   Discussion
Top
 Abstract
 Introduction
 Subjects and Methods
 Results
 Discussion
References
 
Our study shows that carriers of the H1 allele (H1S447 and H1X447 genotypes) present a lower postprandial lipemic response than subjects with the H2S447 genotype (homozygotes for the H2 allele of the LPL HindIII polymorphism and S447 allele).

Several studies have demonstrated the association between the H2H2 genotype and elevated fasting TG levels (12, 13) and LDL-C (13) and lower HDL-C plasma levels (14, 37). However, in this study, we did not observe any difference in basal lipid levels among the three groups of subjects, possibly because our study population was comprised of young, healthy individuals, and the sample size was small, making it difficult to determine differences in transversal studies. An altered lipoprotein phenotype may only become evident when associated with other exogenous factors (13) and aging. Our findings are in accordance with Wilson et al. (38), who have already demonstrated that expression of a partial deficit of LPL is age dependent and that the hypertriacylglycerolemia characteristic of this condition does not usually appear until the fourth decade of life, often coinciding with other secondary factors such as obesity or hyperinsulinemia. In addition, during studies designed to identify the common functional change that explains the HindIII effect, a number of relatively common mutations have been identified, such as D9N and N291S. Although carriers of either of the first two mutations tend to have elevated levels of plasma TG, lower HDL-C levels, and a greater risk of atherosclerosis, their low frequency of occurrence in the population means that they cannot explain the HindIII effect. A third polymorphism has been identified that converts the penultimate amino acid Serine447 to a stop codon (S447X), resulting in a truncation of the enzyme and higher expression in vitro studies. This polymorphism is common, with the allele frequency of the S447X mutation being 20% in healthy individuals. Although early studies examining the association between the S447X polymorphism and lipid levels did not give consistent results, the X447 allele occurs less frequently in patients with hypertriglyceridemia than in healthy whites and is more common in healthy control subjects compared with MI patients.

We demonstrated a delayed postprandial clearance of TRL of intestinal origin in subjects with the H2S447 genotype compared with H1X447 subjects. Because the magnitude of postprandial lipemia is subject to considerable individual variability and is affected by several genetic factors (29, 39, 40, 41, 42), our study sample was restricted to young men with the Apo E3/E3 genotype to avoid possible changes in lipoprotein metabolism caused by other Apo E genotypes (29). In addition, the multiple regression analyses showed an independent effect of LPL polymorphism, excluding the influence of other variables that determine postprandial lipemia response such as BMI, age, TG, HDL-C, and Apo AIV polymorphism. In accordance with our results, a previous study demonstrated that subjects with the H1X447 genotype have a lower postprandial response in plasma TG than H2 subjects (23). However, the lipoprotein fractions involved in this phenomenon were not determined. In our study, we have shown that subjects with H1X447 have the lowest postprandial response due to a lower response of TRL intestinal particles, as reflected in the lower response of Apo B-48 and RP levels. This phenomenon could be involved in the lower risk of CAD associated with this genotype described in previous studies (12, 16, 23).

The catabolism of chylomicrons and chylomicron remnants in the circulation is controlled by a number of factors. Some of these depend on the integrity of the lipolytic and binding functions of the LPL, such as the binding of the chylomicrons to the vascular wall (5), hydrolysis of the TG transported by the particle (1, 2), and hepatic uptake of chylomicron remnants (3). Two mechanisms could possibly explain the change observed in the catabolism of TRL in subjects with LPL polymorphism: a deficit in the catalytic activity of the enzyme (mainly attributed to the amino- terminal functional domain of the mature protein) (1), or an alteration in its binding property (mainly dependent on the carboxy terminal) (43, 44). The LPL HindIII polymorphism is located in intron 8 of the LPL gene (11) and has no direct effect on the amino acid sequence of the mature protein. Its effect on the metabolism of TRL suggests that this polymorphism is a marker of an important functional mutation for this enzyme with which it would be in binding disequilibrium, with repercussions for the catabolism of TRL (45). In addition, the observed effect may also be due to differences in the regulation of the LPL gene between the two genotypes. Insulin is an important regulator of the LPL enzyme at the mRNA and/or posttranscriptional level (46). For example, the H2 allele could be associated with a functional LPL enzyme that is less sensitive to insulin and may result in a reduced response of LPL to insulin in the postprandial period. Other regulators could also be involved in mediating the effects described in this study. The LPL X447 mutation results in a truncation of the enzyme (17) and higher expression in in vitro studies (18). Recent studies have furthermore demonstrated that the X447 mutation is associated with higher postheparin LPL activity (21). This could explain the lower postprandial response observed in subjects with the H1X447 genotype in our study. In addition, the X447 mutation may affect nonenzymatic functions of LPL, such as its bridging function, and may therefore affect postlipolytic clearance of atherogenic lipoprotein remnants. Furthermore, because all the study subjects were young and healthy, it is possible that the relatively small effects on postprandial lipemia response observed in these young healthy individuals will be magnified as the subjects become older or obese, as previously shown with the LPL HindIII polymorphism and basal TG levels (13).

In conclusion, our data suggest that the LPL polymorphism affects the metabolism of TRL during the postprandial period, resulting in prolonged postprandial lipemia in subjects with the H2 allele and a lower postprandial response in subjects with the H1X447 genotype. These phenomena could be involved in the increased prevalence of CAD observed in subjects homozygous for the H2 allele of the HindIII polymorphism (12, 16) and the lower risk of myocardial infarction for the H1X447 genotype (23).


   Footnotes
 
This work was supported by research grants from the CICYT (SAF 96/0060, OLI 96/2146 to F.P.-J., SAF 01/2466-C05 04 to F.P.-J., and SAF 01/03666 to J.L.-M.), the Spanish Ministry of Health (FIS, 98/1531 and 01/0449 to J.L.-M., and FIS 99/0949 to F.P.-J.), Fundación Cultural "Hospital Reina Sofía-Cajasur" (to P.G.), Agencia Española de Cooperación Internacional (to E.P.), Consejería de Salud, Servicio Andaluz de Salud (PAI 97/58, 98/126, 99/116, 00/212, and 01/243 to J.L.-M.; PAI 97/57, 98/132, 99/165, and 00/39 to F.P.-J.; and PAI 01/239 to F.P.-J.), Diputación Provincial de Córdoba (to C.M.), Patrimonio Comunal Olivarero (to F.P.-J.), and The National Institutes of Health, Bethesda, MD (HL 54776 to J.M.O.).

Abbreviations: Apo, Apolipoprotein; AUC, area under the curve; BMI, body mass index; CAD, coronary artery disease; HDL, high-density lipoprotein(s); HDL-C, HDL cholesterol; LDL, low-density lipoprotein(s); LDL-C, LDL cholesterol; LPL, lipoprotein lipase; RP, retinyl palmitate; TG, triacylglycerol(s); TRL, triacylglycerol-rich lipoprotein(s); VLDL, very-low-density lipoprotein(s).

Received September 19, 2003.

Accepted May 20, 2004.


   References
Top
 Abstract
 Introduction
 Subjects and Methods
 Results
 Discussion
 References
 

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