Paraoxonase Gln-Arg(192) and Leu-Met(55) gene polymorphisms and enzyme activity in a population with a low rate of coronary heart disease

doi:10.1016/S0009-9120(02)00295-3

Clinical Biochemistry

Volume 35, Issue 3, May 2002, Pages 197-203

https://doi.org/10.1016/S0009-9120(02)00295-3 Get rights and content

Abstract

Objectives: To assess whether paraoxonase (PON1) polymorphisms at positions 55 and 192 and/or their phenotypic expressions influence the risk of myocardial infarction (MI) in Spanish population.

Design and methods: Two hundred and fifteen male survivors of a MI and their age-matched controls were included in the study. Lipids, apolipoproteins (apo) A-I and B, PON1 activity on paraoxon and phenylacetate and PON1 polymorphisms were determined.

Results: Genotype distribution was similar in patients and controls. Enzyme activities were lower in patients, but multiple logistic regression analysis did not show any independent association with a higher risk of MI.

Conclusion: None of the PON1 polymorphisms or their corresponding measured activities are independent risk factors for MI in our population.

Introduction

Oxidative stress in low-density lipoproteins (LDL) is considered to be a major contributor to the atherogenic process, the principal cause of mortality in developed countries [1]. High-density lipoproteins (HDL) are thought to protect LDL from oxidation [2], [3] due to the presence of antioxidant enzymes that hydrolyze biologically active lipids in oxidized LDL (oxLDL), among which PON1 seems to be of major importance [4], [5], [6], [7]. PON1 is a calcium-dependent esterase, closely associated to HDL in serum, which is known to catalyze the hydrolysis of organophosphates. Its physiologic role, however, has not been totally clarified.

PON1 has two aminoacid polymorphisms. The one at position 192 [glutamine (Q allele)/arginine (R allele)] modulates the hydrolytic activity of PON1 toward some exogenous substrates—such as paraoxon. The activity of the isoenzymes with glutamine at position 192 is lower while the activity of the isoenzymes with arginine at position 192 is increased by high concentrations of salt. However, this polymorphism does not affect the activity toward other substrates, like phenylacetate [8]. The other polymorphism, located at position 55, [leucine (L allele)/methionine (M allele)], does not affect the intrinsic activity of the enzyme so clearly, although it has been shown to modulate its concentration in plasma [9], [10].

Some studies have associated the presence of the R allele or the homozygosity of the L allele with a greater susceptibility to coronary heart disease (CHD) [11], [12], [13], [14], [15], [16], [17] but this is still controversial [18], [19], [20], [21], [22], [23], [24], [25], [26], [27], [28]. More recently, the PON1 activity phenotype has been shown to be a better predictor of CHD than the PON1 (192) or PON1 (55) genotypes [29]. Our aim was to assess the relationship between PON1 activity and both of these gene polymorphisms in the Spanish population, which has one of the lowest incidences of MI in Western societies [30].

Section snippets

Subjects

The procedures complied with the ethical standards of the Hospital Universitari de Sant Joan and the Hospital Universitari Joan XXIII. As in a previous study [31], we located the medical records of patients registered as having had a MI and invited the surviving male patients who had a history such as that defined by the criteria of the World Health Organization to participate [32]. We made a baseline examination and assessment of the cardiovascular status of the first 215 patients who agreed

Study population and cardiovascular risk factors

Patients with MI were relatively young and the serum concentrations of HDL cholesterol and apo A-I were lower than in controls. The serum concentrations of cholesterol, triglycerides and apo B were higher in patients than in controls (Table 1). As expected, the prevalence of diabetes, hypertension and cigarette smoking was also higher in patients than in controls. Arylesterase activity was similar in patients and in controls but the paraoxonase activity (baseline and stimulated) was

Discussion

There is increasing evidence showing that serum PON1 is related to the prevention of CHD [11], [12], [13], [14], [15], [16], [17]. According to the oxidation hypothesis of atherosclerosis, this enzyme prevents lipoperoxides from accumulating in LDL particles [3], [4], [5]. Consequently, considerable research has been done to determine its physiologycal role and the functional differences between its isoenzymes. The Gln-Arg192 and Leu-Met55 polymorphisms are known to affect PON1 activity using

Acknowledgements

Supported by grants 98/0425, 99/0901 and 00/0291 from the Fondo de Investigaciones Sanitarias. N. Ferré was supported by grant FI/FIAP 99 from the Generalitat de Catalunya.

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There is an increasing prevalence of coronary artery disease (CAD) in younger individuals. Lipid biomarkers such as lipoprotein-a (Lp-a), Apo A1, Apo B and Paraoxonase-1 (PON1) serve as important risk predictors for development of CAD. There is little evidence regarding the role of lipid biomarkers and their genetic polymorphisms in young (<50 years) ST-segment elevation myocardial infarction (STEMI) patients.
This study included 110 young (18–50 years) STEMI patients and 110 healthy controls. Serum levels of Apo A1, Apo B, Paraoxonase-1 (PON-1) and Lipoprotein-associated phospholipase A2 (Lp-PLA2) were estimated for both patients as well as controls. Additionally, genetic polymorphisms in the Apo A1 (75G/A) and the PON1 (Q192R) genes were evaluated.
Serum levels of apo B (101.31 ± 27.58 vs 75.31 ± 18.77 mg/dl; p < 0.0001), Lp(a) [87.56 ± 74.28 vs 25.81 ± 24.66 mg/dl, p < 0.0001] and Lp-PLA2 [5.97 ± 1.39 vs 3.49 ± 1.27 ng/mL, p < 0.0001] were significantly higher in patients as compared to controls. Serum levels of Apo A1 [44.76 ± 35.65 vs 95.97 ± 29.89; p < 0.0001] and PON1 [2.63 ± 1.5 vs 3.87 ± 1.47 ng/mL, p < 0.0001] were significantly lower in cases as compared with controls. Additionally, patients with genetic polymorphisms in the Apo A1 (75G/A) and the PON1 (Q192R) gene had an increased risk of STEMI.
Lipid biomarkers such as Apo A1, Apo B and PON1 and their genetic polymorphism are associated with the susceptibility for STEMI in young individuals.
Association between PON1 activity and coronary heart disease risk: A meta-analysis based on 43 studies
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Paraoxonase 1 (PON1) is reported to have antioxidant and cardioprotective properties. The relationship between PON1 activity and coronary heart disease (CHD) risk in humans has been reported among various ethnic populations in the past decade. However, these studies have yielded contradictory results. To investigate this inconsistency, we conducted a meta-analysis of 43 studies involving a total of 20,629 subjects to evaluate the effect of PON1 activity on susceptibility for CHD. We also systematically explored potential sources of heterogeneity using subgroup analysis and meta-regression. Significant decreases paraoxonase activity of PON1 were observed in CHD patients compared with non-CHD controls with SMD of − 0.78 (95% CI: −0.98, −0.57; P < 0.001). Similar results were also found for arylesterase activity of PON1 with SMD of − 0.50 (95% CI: −0.64, −0.36; P < 0.001). In the subgroup analysis by ethnicity, CHD phenotype, sample size, source of controls, mean age and BMI of cases, significantly increased risks were also found. In addition, our analyses detected a possibility of publication bias with an overestimate of the true association by smaller studies. This meta-analysis demonstrated that decreasing in PON1 activity is a risk factor associated with increased CHD susceptibility. However, additional very large-scale studies are warranted to provide conclusive evidence on the effects of PON1 activity on risk of CHD.
Four genetic polymorphisms of paraoxonase gene and risk of coronary heart disease: A meta-analysis based on 88 case-control studies
2011, Atherosclerosis
The human paraoxonase (PON) is calcium dependent HDL associated ester hydrolase which has attracted considerable attention as a candidate gene for coronary heart disease based on its enzyme function as a key factor in lipoprotein catabolism pathways. Many studies have examined the association between polymorphisms in the PON gene and risk of coronary heart disease (CHD), but the results have been inconsistent.
We conducted a meta-analysis of 88 studies on 4 PON polymorphisms [Q192R, L55M, and T(−107)C in the PON1 and the S311C in the PON2] published before August 2010, including a total of 24,702 CHD cases and 38,232 controls. We also systematically explored potential sources of heterogeneity.
In a combined analysis, the summary per-allele odds ratio for CHD of the 192R was 1.11 (95% CI: 1.05–1.17). However, when the analyses were restricted to 10 larger studies (n > 500 cases), the summary per-allele odds ratio was 0.96 (95% CI: 0.90–1.02). Our analyses detected a possibility of publication bias with an overestimate of the true association by smaller studies. A meta-analysis of studies on the 55M, (−107)T, and 311C variant showed no significant overall association with CHD, yielding a per-allele odds ratio of 0.94 (95% CI: 0.88–1.00), 1.02 (95% CI: 0.91–1.15) and 1.02 (95% CI: 0.90–1.16) respectively.
This meta-analysis suggested an overall weak association between the R192 polymorphism and CHD risk.
Oxidized to non-oxidized lipoprotein ratios are associated with arteriosclerosis and the metabolic syndrome in diabetic patients
2008, Nutrition, Metabolism and Cardiovascular Diseases
Type 2 diabetic patients have a greater prevalence of the metabolic syndrome, oxidative stress and accelerated atherosclerosis, compared to non-diabetics. We examined the association between biomarkers of lipid peroxidation and the presence of atherosclerosis and the metabolic syndrome in diabetic patients.
We studied oxidized LDL (OxLDL), OxLDL/LDL, OxLDL/HDL, lipoperoxides, autoantibodies against OxLDL (OxLDL-Ab), diene formation of LDL (lag phase), vitamin E, vitamin E/cholesterol and PON1 polymorphisms (−108C > T, 55T > A, and 192A > G) in 166 non-smoking type 2 diabetic patients, 119 fulfilling the criteria for the metabolic syndrome, 73 with atherosclerosis and 93 without atherosclerosis. Patients with macrovascular disease had higher values of OxLDL/LDL (11%; P = 0.016), OxLDL/HDL (18%; P = 0.024) and OxLDL-Ab (12%; P = 0.046). OxLDL/LDL and OxLDL/HDL were correlated with the number of components of the metabolic syndrome (P < 0.001). PON1 polymorphisms were not associated to LDL oxidation markers, only PON1 (−108TT) was weakly associated with higher OxLDL-Ab concentrations (22%; P = 0.040) in patients with atherosclerosis.
OxLDL/LDL, OxLDL/HDL and OxLDL-Ab are the most useful clinical parameters of lipoprotein oxidation for discriminating the presence of macrovascular disease in diabetic patients. The presence of the metabolic syndrome in these patients is also associated with an increase in the oxidized lipoprotein ratios.
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In recent years, importance of enzyme activity measurements, in addition to genotyping, in epidemiological studies relating paraoxonase 1 (PON1) and vascular disease was emphasized. This is the first report evaluating paraoxonase and arylesterase activities as risk factors for ischemic stroke. In addition, PON1 192Gln(Q)/Arg(R) and 55Leu(L)/Met(M) polymorphisms were also analyzed.
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Citation Excerpt :
The intragenotype variability (4- to 8-fold) is significantly larger than the mean intergenotype differences (1.2- to 1.5-fold); thus, even in the small sample examined here, we could observe many individuals with the inferior QQ genotype that exhibit higher levels of HDL-associated PON1, and subsequently higher lipolactonase activity, than most RR individuals. Numerous case-control studies that tried to relate the PON1 R/Q polymorphism with the risk of cardiovascular disease yielded conflicting results, with some studies indicating the RR genotype as a risk factor (42–44) and others indicating no association between the disease and either allele (45–48). Our tests show that the PON1 R/Q polymorphism plays a relatively minor role in determining the levels of the HDL-PON1 complex and may explain why previous attempts to correlate the 192R/Q phenotype with a predisposition for atherosclerosis failed.
Serum paraoxonase (PON1) is a lipolactonase that associates with HDL-apolipoprotein A-I (HDL-apoA-I) and thereby plays a role in the prevention of atherosclerosis. Current sera tests make use of promiscuous substrates and provide no indications regarding HDL-PON1 complex formation. We developed new enzymatic tests that detect total PON1 levels, irrespective of HDL status and R/Q polymorphism, as well as the degree of catalytic stimulation and increased stability that follow PON1's tight binding to HDL-apoA-I. The tests are based on measuring total PON1 levels with a fluorogenic phosphotriester, measuring the lipolactonase activity with a chromogenic lactone, and assaying the enzyme's chelator-mediated inactivation rate. The latter two are affected by tight HDL binding and thereby derive the levels of the serum PON1-HDL complex. We demonstrate these new tests with a group of healthy individuals (n = 54) and show that the levels of PON1-HDL vary by a factor of 12. Whereas the traditionally applied paraoxonase and arylesterase tests weakly reflect PON1-HDL levels (R = 0.64), the lipolactonase test provides better correlation (R = 0.80). These new tests indicate the levels and activity of PON1 in a physiologically relevant context as well as the levels and quality of the HDL particles with which the enzyme is associated.

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ClinicalParaoxonase Gln-Arg(192) and Leu-Met(55) gene polymorphisms and enzyme activity in a population with a low rate of coronary heart disease

Abstract

Introduction

Section snippets

Subjects

Study population and cardiovascular risk factors

Discussion

Acknowledgements

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Clinical
Paraoxonase Gln-Arg(192) and Leu-Met(55) gene polymorphisms and enzyme activity in a population with a low rate of coronary heart disease