同型半胱氨酸硫内酯-同型半胱氨酸致血管病变的基础

同型半胱氨酸硫内酯是氨基酰-tRNA合成酶在编辑或校正过程中形成的同型半胱氨酸反应产物,它可直接或通过蛋白质同型半胱氨酸化损伤内皮细胞。而内皮细胞同型半胱氨酸硫内酯的合成及蛋白质同型半胱氨酸化程度取决于同型半胱氨酸/蛋氨酸、叶酸、高密度脂蛋白等。同型半胱氨酸硫内酯解酶作为体内降解同型半胱氨酸硫内酯的惟一酶类,其活性的高低在与同型半胱氨酸相关的血管病变中可能起重要作用。     

Relation between butyrylcholinesterase K variant,paraoxonase 1 (PON1) Q and R and apolipoprotein E epsilon 4 genes in early-onset coronary artery disease

OBJECTIVES: The common K variant of butyrylcholinesterase (BChE-K), an enzyme which metabolizes acetylcholine and organophosphates, has been associated with Alzheimer's disease, especially in the presence of the apolipoprotein E epsilon 4 allele (APOE-epsilon 4). Although APOE-epsilon 4 has been associated with the development of coronary artery disease (CAD), an association between the BChE-K variant and CAD has not been explored. Paraoxonase 1 (PON1), located within HDL, is an enzyme which also metabolizes organophosphates and may be antiatherogenic. The R192 variant of PON1 (PON1-R) has been associated with CAD. DESIGN AND METHODS: To determine whether BChE-K is also associated with premature CAD, we examined the frequency of BChE-K among patients with early-onset CAD (n = 150; < 50 yr) vs. late-onset CAD (n = 150; > 65 yr) by molecular analysis. We also examined the frequency of the PON1-R allele in both groups, and explored whether there was synergism between BChE-K and APOE-epsilon 4, BChE-K and PON1-R or PON1-R and APOE-epsilon 4. RESULTS: The frequency of the BChE-K allele tended to be greater among early-onset CAD patients compared to late-onset CAD patients (41.3% vs. 31.3%; p = 0.07), but without any significant difference between males and females. There was no difference in the prevalence of the PON1-R allele between those with early- or late-onset CAD (46.0% vs. 52.7%; p = 0.25). Twenty-two patients with early-onset CAD had both the BChE-K plus APOE-epsilon 4 alleles (14.7%) compared to 11 late-onset CAD patients (7.3%) (p = 0.04). There was no such association between BChE-K and PON1-R, nor PON1-R and APOE-epsilon 4.CONCLUSIONS: Our study suggests that there is a minor association between BChE-K and early-onset CAD, especially in the presence of the APOE-epsilon 4 allele.     

Serum paraoxonase activity in patients with type 1 diabetes compared to healthy controls

BACKGROUND: The oxidation of low-density lipoprotein (LDL) is central to current theories on the initiation and progression of atherosclerosis. Type 1 diabetes is associated with an increase in oxidative stress, which may be responsible for the increased susceptibility to coronary heart disease seen in type 1 diabetes. High-density lipoprotein (HDL) associated paraoxonase (PON1) can retard the oxidation of LDL. DESIGN: Paraoxonase activity, concentration and genotype were therefore investigated in 152 people with type 1 diabetes and 282 healthy controls. These parameters were also investigated in the group with type 1 diabetes in relation to the presence of diabetic complications. RESULTS: Both PON1 activity and concentration were significantly lower by 16.7% and 19.2% (both P < 0.05) in the type 1 diabetes group. These differences were independent of the PON1 coding region polymorphisms. The distribution of PON1 activity and mass were the same in both populations, i.e. for the PON1-192 polymorphism RR > RQ > QQ and for the PON1-55 polymorphism LL > LM > MM. There were no differences in either the PON1 polymorphisms, PON1 activity and concentration in people with type 1 diabetes in the presence or absence of micro and macro vascular complications of diabetes. CONCLUSIONS: Low PON1 activity may contribute to the increased atherosclerosis found in type 1 diabetes by reducing the ability of HDL to retard LDL oxidation despite the frequently-found increased HDL in type 1 diabetes when good glycaemic control is established.     

Paraoxonase gene polymorphism, serum lipid, and oxidized low-density lipoprotein in preeclampsia

OBJECTIVES: Human paraoxonase-1 (PON-1) is thought to play a role in preeclampsia and atherosclerosis, mainly through a reduction in low-density lipoprotein oxidation. Oxidized low-density lipoprotein (LDL) is very important in endothelial dysfunction of preeclampsia. The aim of the present study was to investigate the association between PON1 gene polymorphism and preeclampsia and to determine concentrations of serum lipid in preeclampsia patients. We aimed also to evaluate serum oxidized LDL levels in normal and preeclampsia patients. STUDY DESIGN: We performed the present study in 57 control women and 32 preeclampsia patients. PON-1 genotypes were determined by polymerase chain reaction amplification and restriction fragment length polymorphism. Serum triglyceride, cholesterol, high-density lipoprotein cholesterol, and low-density lipoprotein cholesterol levels were measured. We also measured serum levels of oxidized LDL by ELISA method. RESULTS: There was no significant difference in PON1 genotype frequencies between the control and preeclampsia patients. The levels of serum cholesterol and high-density lipoprotein were significantly lower in preeclampsia patients compared with that of the control women (p=0.05, and p<0.01, respectively). The serum levels of oxidized LDL in preeclampsia patients were significantly higher than those in the control women (p=0.001). CONCLUSIONS: These findings support the importance of the atherogenic lipid profile and oxidized LDL that is enhanced in preeclampsia, and these findings may be significant contributors to endothelial dysfunction.     

Paraoxonase-1 activity as a marker of atherosclerosis is not associated with low bone mineral density in healthy postmenopausal women

The aging process is associated with an increasing prevalence of osteoporosis and atherosclerosis, but it is uncertain if these two conditions are interrelated. Serum paraoxonase-1 (PON1) is a high-density lipoprotein (HDL) associated enzyme that has been implicated in the pathogenesis of atherosclerosis. Our aims of the study were to investigate (1) serum paraoxonase and arylesterase activities and, lipid hydroperoxide (LOOH) levels in healthy postmenopausal women and (2) whether there were any associations between these enzyme activities and bone mineral density (BMD). A total of 97 generally healthy postmenopausal women were enrolled in the study. BMD was measured at lumbar spine (LS) and femoral neck (FN) with dual energy X-ray absorptiometry. Serum paraoxonase and arylesterase activities were measured spectrophotometrically. LOOH levels were measured by iodometric assay. In this population, 50 (51%) women had BMD T scores < −2.5 at the LS and/or FN defined as osteoporosis and 47 (49%) of them had normal BMDs. Serum paraoxonase, arylesterase, and LOOH activities were not significantly different between osteoporotic and nonosteoporotic postmenopausal women. There were also no correlations between paraoxonase, arylesterase, LOOH activities, and LS BMD and FN BMD. We conclude that there may be not good evidence to support a direct relationship between osteoporosis and atherosclerosis in these subjects. However, prospective studies with larger groups are needed to investigate this issue further.     

The inhibition effects of some sulfonamides on human serum paraoxonase-1 (hPON1)

BackgroundParaoxonase 1 (PON1) is an antiatherogenic and organophosphate hydrolyzer enzyme. It has important roles including protecting low density lipoprotein (LDL) against oxidation and the detoxification of highly toxic substances. Reducing the levels of this enzyme in patients with diabetes mellitus, cardiovascular diseases, hyperthyroidism, and chronic renal failure is a major risk.MethodsHere, we report on the purification of the human serum PON1 using simple methods and determine the interactions between some sulfonamides and the enzyme.ResultsWe found that some sulfonamides exhibit potential inhibitor properties for the human serum PON1 with IC₅₀ values in the range of 24.10–201.45 μM and K_i values in the range of 4.41 ± 0.52–150.23 ± 20.73 μM. The sulfonamides showed different inhibition mechanisms. We determined that sulfonamides 1, 2, 4, 5, 8, and 9 showed a non-competitive inhibition effect whereas sulfonamides 3, 6 and 7 showed competitive inhibition.ConclusionUse of drugs containing the sulfonamides molecule groups with crucial biological activity would be very dangerous in some cases.     

Butyrylcholinesterase, paraoxonase, and albumin esterase, but not carboxylesterase, are present in human plasma

The goal of this work was to identify the esterases in human plasma and to clarify common misconceptions. The method for identifying esterases was nondenaturing gradient gel electrophoresis stained for esterase activity. We report that human plasma contains four esterases: butyrylcholinesterase (EC 3.1.1.8), paraoxonase (EC 3.1.8.1), acetylcholinesterase (EC 3.1.1.7), and albumin. Butyrylcholinesterase (BChE), paraoxonase (PON1), and albumin are in high enough concentrations to contribute significantly to ester hydrolysis. However, only trace amounts of acetylcholinesterase (AChE) are present. Monomeric AChE is seen in wild-type as well as in silent BChE plasma. Albumin has esterase activity with alpha- and beta-naphthylacetate as well as with p-nitrophenyl acetate. Misconception #1 is that human plasma contains carboxylesterase. We demonstrate that human plasma contains no carboxylesterase (EC 3.1.1.1), in contrast to mouse, rat, rabbit, horse, cat, and tiger that have high amounts of plasma carboxylesterase. Misconception #2 is that lab animals have BChE but no AChE in their plasma. We demonstrate that mice, unlike humans, have substantial amounts of soluble AChE as well as BChE in their plasma. Plasma from AChE and BChE knockout mice allowed identification of AChE and BChE bands without the use of inhibitors. Human BChE is irreversibly inhibited by diisopropylfluorophosphate, echothiophate, and paraoxon, but mouse BChE spontaneously reactivates. Since human plasma contains no carboxylesterase, only BChE, PON1, and albumin esterases need to be considered when evaluating hydrolysis of an ester drug in human plasma.     

The paraoxonase promoter polymorphism (-107)T>C is not associated with carotid intima-media thickness in Sicilian hypercholesterolemic patients

BACKGROUND AND OBJECTIVES: Increased plasma low-density lipoprotein-cholesterol (LDL-C) levels in hypercholesterolemic subjects are associated with enhanced LDL oxidation that represents an additional risk for atherosclerotic disease. Human serum paraoxonase (PON1), a high-density lipoprotein (HDL) associated enzyme, has been shown to protect LDL from oxidation, thus playing an important role in reducing the risk of atherosclerosis. PON1 gene polymorphisms have been found to be associated with the variations in serum PON1 levels and activities, and with the risk for coronary artery disease (CAD). This study was performed to evaluate the contribution of the PON1 promoter (-107)T>C and the coding region Gln 192 Arg (Q192R) and Leu 55 Met (L55M) polymorphisms to the presence of carotid atherosclerosis in 208 Sicilian subjects with primary hypercholesterolemia. METHODS: Carotid artery intima-media wall thickness (IMT) was measured as an indicator of early atherosclerotic disease. The subjects were classified according to whether they have a normal (1 mm) IMT. Subjects were also investigated for physical and biochemical parameters, including PON1 activity. RESULTS: No significant differences were detected among the PON1 genotypes with respect to age, sex, BMI, plasma lipids, systolic blood pressure in both groups of patients. There were significant differences between PON1 genotypes with respect to PON1 activity. The 192QQ, 55MM and (-107)TT genotypes showed lower PON1 activity compared to the RR, LL and CC genotypes. The PON1 (-107)T>C genotype distribution in both IMT groups showed no significant differences in percentage of TT, CT and CC genotypes. Similar results were obtained analyzing the Q192R and L55M genotype frequencies. Stepwise forward logistic regression analysis confirmed the lack of association between PON1 genotypes and carotid abnormalities. CONCLUSIONS: In conclusion, our data provided no evidence of a significant association between either PON1 promoter (-107)T>C or coding region, Q192R and L55M, polymorphisms and early carotid atherosclerosis in Sicilian hypercholesterolemic subjects.