中国老年人心肌梗死遗传易感性与血管内皮型一氧化氮合酶基因Glu298Asp多态性的关系

背景诸多研究认为血管内皮型一氧化氮合酶(endothelial nitric oxide synthase,eNOS)基因多态性与心、脑、肾疾病存在相关性,但eNOS-Glu298Asp与中国老年人心肌梗死的关系是否相关尚不明确.设计以诊断为依据的病例对照研究.目的探讨老年人血管内皮型一氧化氮合酶基因Glu298Asp多态性的分布,及其与老年心肌梗死的关系.地点、对象和方法37例老年心肌梗死患者为病例组,均为南京医科大学第一附属医院门诊及住院患者,其中20例既往无高血压史者为病例亚组;172例本院体检中心复检者(对照组),其中92例既往无高血压者为对照亚组,分别检测病例组和对照组的身高、体质量、空腹血脂等指标,采用聚合酶链反应(polymerase chain reaction,PCR)和限制性片断长度多态性(restrictionfragmentlength polymorphism,RFLP)检测eNOS基因Glu298Asp多态性.主要观察指标4组老年人eNOS基因Glu298Asp多态性及298Asp等位基因频率.结果病例组Glu/Asp基因型高于对照组(32.43%和18.02%),两组eNOS基因Glu298Asp多态性构成差异有显著性意义(x2=3.87,P<0.05).病例亚组Glu/Asp基因型(35.00%)高于对照亚组(10.87%),差异有显著性意义(x2=7.43,P<0.01).病例组298Asp等位基因频率高于对照组,差异无显著性意义(P>0.05).病例亚组298Asp等位基因频率高于对照亚组,差异有显著性意义(x2=6.82,P<0.01).结论eNOS基因Glu298Asp多态性在中国老年人群中存在,Glu/Asp基因型和298Asp等位基因可能是中国老年人心肌梗死的遗传易感指标.     

Gender specific association of endothelial nitric oxide synthase gene (Glu298Asp) polymorphism with essential hypertension in a south Indian population

BACKGROUND: Endothelial derived nitric oxide (NO) plays a major role in blood pressure regulation. The role of missense variant eNOS-Glu298Asp has been demonstrated by many studies with conflicting results. Our objective was to investigate the association of eNOS gene polymorphism with essential hypertension in a south Indian population. METHODS: We carried out a case control study in 438 hypertensive patients and 444 healthy control subjects in a homogenous population. Genotyping was done by PCR-RFLP method. Multiple logistic regression analysis was used to detect the association between genotype and hypertension. RESULTS: The homozygous variant genotype Asp298Asp was significantly associated with hypertension (odds ratio 2.4; 95% CI, 1.23-5.0, p<0.01). Gender specific analysis showed both the heterozygous (odds ratio, 2.0; 95% CI, 1.3-2.9, p<0.01) and homozygous variants (odds ratio, 7.9; 95% CI, 2.0-4.1, p<0.001) were positively associated with hypertension in females. The variant allele Asp was higher in female hypertensives when compared to male hypertensive cases (22% vs. 16%). CONCLUSION: The eNOS gene polymorphism is a candidate gene for hypertension and the association to be gender specific with respect to females in a south Indian Tamilian population.     

Endothelial nitric oxide synthase (Glu298Asp) polymorphism is an independent risk factor for migraine with aura

OBJECTIVE: The aim of the present study was to evaluate whether the functional endothelial nitric oxide synthase (eNOS) Glu298Asp polymorphism, which has been demonstrated to decrease the endothelial NOS activity, might be a risk factor for migraine. BACKGROUND: It has widely demonstrated that nitric oxide (NO) is involved in migraine pathogenesis. Several genetic risk factors have been associated with migraine, but no study has unraveled a possible relationship between migraine and eNOS Glu298Asp. Methods.-One hundred fifty-six migraine patients and 125 healthy nonheadache volunteers entered the study. Demographic and clinical characteristics were carefully recorded, and a neurological workup was performed. RESULTS: eNOS AspAsp homozygous patients had a 3-fold time risk for migraine with aura (MA) when compared to migraine without aura (MO) patients (OR-3.02, 95% CI-1.21 to 7.51), and more than 2-fold time increased risk when compared to control subjects (OR-2.21, 95% CI-1.00 to 5.04). In migraine patients, no difference in age at onset, mean attack's intensity, family history for any of the studied comorbidities, or the presence of comorbidities was found in eNOS AspAsp homozygous compared to eNOS GluGlu or eNOS GluAs carriers. CONCLUSIONS: Homozygous Asp298, a common variant of the eNOS gene, is an independent risk factor for MA in this study population.     

Endothelial nitric oxide synthase Glu298Asp gene polymorphism influences body composition and biochemical parameters but not the nitric oxide response to eccentric resistance exercise in elderly obese women

Both endothelial nitric oxide synthase (eNOS) gene polymorphism and nitric oxide (NO) are involved in important cardiovascular, muscular and inflammatory physiological mechanisms during ageing and response to exercise. The aim of this study was to investigate the NO kinetic response following an acute eccentric resistance exercise (ERE) session and the possible effect of the Glu298Asp eNOS gene polymorphism in elderly obese women. Eighty‐seven women (age 69·4 ± 6·1 years, body weight 74·9 ± 12·7 kg, height 151·9 ± 6·0 cm and BMI 32·5 ± 5·7 kg m^?2) completed seven sets of ten eccentric repetitions at 110% of the ten repetitions maximum (10RM). NO concentrations remained elevated up to 48 h following the acute ERE session as compared with baseline, for GG and GT/TT groups (P<0·05), with no differences between genotypes. The GG genotype group had higher body weight, prevalence of obesity (BMI classification – 81% versus 56%), BMI and higher relative muscle strength, while they had significantly lower triglycerides, VLDL and urea concentrations as compared with TT/TG group. In conclusion, NO remains elevated for up to 48 h after an acute ERE session, without genotype interaction. The TT/TG genotype had a negative impact on triglycerides, VLDL and urea concentrations. Thus, T carriers should increase their attention to cardiovascular risk factor and metabolic disorders.     

The Glu298ASP polymorphism of the endothelial nitric oxide synthase gene is associated with premature ST elevation myocardial infarction in Mexican population

BackgroundThe polymorphism Glu298Asp of endothelial nitric oxide (eNOS) gene has been associated with hypertension and coronary artery disease in several populations worldwide, but results are still controversial. We examined the possible association of the Glu298Asp with premature ST elevation myocardial infarction (STEAMI) in young Mexican population.MethodsIn a case–control study 180 unrelated patients with STEAMI ≤ 45 years who were admitted to a cardiovascular intense care unit and 180 apparently healthy controls matched by age and gender were recruited from January 2006 to June 2009. The polymorphism Glu298Asp was determined in all participants by a polymerase chain-reaction-restriction fragment length polymorphism assay (PCR-RFLP).ResultsThere was a significant difference in the genotype distribution between 2 groups (P = 0.001). The allele Asp occurred more frequently in the patients group (P = 0.001). There were independent factors for STEAMI: the Asp allele (OR 2.2, 95% CI 1.1–3.5, P = 0.03), smoking (OR 5.0, 95% CI 3.1–8.2, P < 0.001), hypertension (OR 2.0, 95% CI 1.0–3.5, P = 0.03), family history of cardiovascular disease, (OR 3.7, 95% CI 2.0–4.6, P = 0.02), and dyslipidemia (OR 3.4, 95% CI 2.0–6.3, P = 0.02).ConclusionsThe Asp allele from the Gu298Asp polymorphism represents an independent risk factor for premature STEAMI in Mexican Mestizo population.     

Lack of association of the Glu298Asp polymorphism of endothelial nitric oxide synthase with manifest coronary artery disease,carotid atherosclerosis and forearm vascular reactivity in two Austrian populations

OBJECTIVE: Conflicting data exists about the possible contribution of the homozygous Asp/Asp genotype of the Glu298Asp polymorphism of endothelial nitric oxide synthase to human atherosclerotic vascular disease. We investigated the polymorphism in two independent study populations: a case-control study including patients with angiographically verified coronary artery disease (CAD) on the one hand and a cross-sectional epidemiological study on the other hand. METHODS: The Glu298Asp polymorphism was determined by PCR-RFLP as established. In the case-control study (240 patients and 248 controls) a possible association between the polymorphism and CAD, and age of onset of CAD and myocardial infarction was investigated. In the cross-sectional epidemiological study (932 subjects) intima-media thickness (IMT) of the carotid artery as well as morphological plaque burden and forearm vascular reactivity (peak postischemic reactive hyperaemia, determined by venous occlusion plethysmography) were measured. RESULTS: In the case-control study genotype distribution (Glu/Glu; Glu/Asp; Asp/Asp) was not different between the CAD patients (43/46/11%) and the controls (49/41/10%, P = NS). No association of the polymorphism with age of onset of CAD or myocardial infarction was found. In the epidemiological study no influence of the genetic variant on IMT was observed after correction for classical determinants of IMT (average IMT: Asp/ Asp: 0.077 +/- 0.011 mm; Glu/Glu and Glu/Asp: 0.080 +/- 0.012 mm, P = NS). Forearm vascular reactivity was also not different between homozygous Asp/Asp subjects and Glu/Glu and Glu/Asp subjects (peak-reactive hyperaemia 20.1 +/- 7.3 mL min-1 100 mL-1 vs. 20.0 +/- 6.5 mL min-1 100 mL-1, P = NS). CONCLUSIONS: Our results suggest that there is no association of the Glu298Asp polymorphism with coronary or carotid atherosclerosis or forearm vascular reactivity in these populations recruited in a country with a rather high risk for atherosclerosis. We suggest additional investigations to be performed in populations at different risk for coronary events to further elucidate the possible contribution of this polymorphism to vascular disease.     

Genetic risk factors for coronary artery spasm: significance of endothelial nitric oxide synthase gene T-786-->C and missense Glu298Asp variants.

BACKGROUND: We recently identified two endothelial nitric oxide synthase (eNOS) gene polymorphisms, Glu298Asp and T-786-->C, which are independently associated with coronary spasm. eNOS gene intron 4b/a polymorphism is also reported to be involved in smoking-dependent coronary artery disease. The genetic linkage among these polymorphisms remains unknown. Also, it is unclear which variant is most responsible for coronary spasm. In the present study, we first examined the genetic linkage among these three variants. Next, we studied the risk factors of coronary spasm by using all significant genetic and conventional risk factors in a large-scale study. METHODS: The genotype and allele frequencies for the T-786-->C, intron 4b/a, and Glu298Asp variants were assessed in 423 randomly selected DNA samples to examine their genetic linkages. The relative capacities of all risk factors to predict coronary spasm were then analyzed using multiple logistic regression in 201 patients with coronary spasm and 345 volunteers. RESULTS: Comparison of allele frequencies revealed that the eNOS intron 4a allele was significantly linked to the T-786-->C mutation (P < 0.00001), whereas there was not a linkage between the intron 4a allele and the Glu298Asp variant (P = 0.1437) or between the Glu298Asp variant and the T-786-->C mutation (P = 0.1996). Multiple logistic regression revealed that the most predictive independent risk factor for coronary spasm was the T-786-->C mutation (P < 0.001), followed by cigarette smoking (P < 0.001), hypertension (P = 0.004), and the Glu298Asp variant (P = 0.028). CONCLUSIONS: We found that the T-786-->C mutation and the intron 4a allele are in linkage disequilibrium. We previously showed that the T-786-->C mutation reduced eNOS gene promoter activity. In that context, our results strongly suggest that the T-786-->C mutation underlies the functional characteristics of the intron 4a allele. Further, multiple logistic regression analysis revealed that the T-786-->C mutation is the most predictive risk factor for coronary spasm, followed by cigarette smoking. Given that those effects are potentially additive, patients carrying the eNOS gene variants should be strongly cautioned against smoking.     

Endothelial nitric oxide synthase Glu(298)-->Asp polymorphism, carotid atherosclerosis and intima-media thickness in a general population sample

The Glu(298)-->Asp (E298D; 894G-->T) polymorphism of eNOS (endothelial nitric oxide synthase) has been related with cardiovascular disease. In the present study, we investigated the association of Glu(298)-->Asp with atherosclerotic plaques in different carotid vessel segments and with carotid IMT (intima-media thickness). The Glu(298)-->Asp eNOS polymorphism was determined by 5'-exonuclease assay among 2448 participants of the SHIP (Study of Health in Pomerania). Mean and maximum common carotid IMT, as well as carotid atherosclerosis, were measured by high-resolution ultrasound. The Asp/Asp(298) genotype was associated with an increased risk of atherosclerotic plaques at the level of the common carotid arteries [multivariate odds ratio, 1.57 and 95% CI (confidence interval), 1.05-2.34; P=0.025], but not in the carotid bifurcations or internal or external carotid arteries. Glu(298)-->Asp genotype was not associated with carotid IMT in the whole sample. However, the Asp/Asp(298) genotype was independently associated with both higher mean [adjusted increase by 0.046 mm (95% CI, 0.013-0.078); P=0.006] and maximum carotid IMT [0.137 mm (95% CI, 0.064-0.209); P<0.001] in the low-risk group of subjects without carotid atherosclerosis. In conclusion, the Asp/Asp(298) genotype is associated with atherosclerosis in the common carotid arteries and, in a low-risk group, also with carotid IMT. This suggests that the association of the Glu(298)-->Asp genotype with atherosclerosis in the carotid arteries is site-specific and is modified by overall cardiovascular risk.     

Association of eNOS Glu298Asp gene polymorphism with essential hypertension in Asian Indians

BACKGROUND: Several reports suggested association between the T allele of the endothelial nitric oxide synthase gene polymorphism (eNOS) Glu298Asp (G --> T at nucleotide 894, exon 7) with essential hypertension (EHT). Findings, however are not uniform. In this case-control study, we determined the prevalence and distribution of the above polymorphism and their relationship with the disease to elucidate the association of this polymorphism with the risk and pathophysiology of EHT in the Asian Indians and significance of T allele in this context. METHODS: Two hundred normal controls and 226 patients with EHT from Delhi and surrounding areas were recruited for the investigation. Venous blood samples were used for genotyping. Genotyping was done by PCR-RFLP. RESULTS: The data suggested higher prevalence of GT+TT genotypes in patients as compared to the controls and association of T allele with EHT [p<0.001, odds ratio at 95% CI, 2.10 (1.34-3.28)]. CONCLUSIONS: Asian Indians of this region with T allele may be at higher risk of EHT.     

内皮一氧化氮合酶基因多态性与糖尿病肾病关系的研究

目的 探讨内皮一氧化氮合酶(eNOS)基因内含子4的多态性与糖尿病肾病的关系,以及其在中国人、马来西亚人和印度人中的分布。方法 选择258例病程在10年以上的2型糖尿病患者作为观察对象,其中中国人150例、马来西亚人71例、印度人37例。从患者全血中提取DNA,然后,进行聚酶链反应(PCR)、克隆及测序。结果 以往认为该基因有2种等位基因(a和b),而本研究发现3种等位基因,并对第3种多态(等位基因c)基因进行了序列分析;统计分析显示,等位基因a、b和c均与糖尿病肾病无显著相关性。结论 eNOS基因内含子4上的多态性有3种等位基因;该基因多态性与糖尿病肾病无显著相关性。     

Hyperhomocysteinemia, endothelial nitric oxide synthase polymorphism, and risk of coronary artery disease

BACKGROUND: Hyperhomocysteinemia is an independent, graded risk factor for coronary artery disease (CAD). The G894T variant of endothelial nitric oxide synthase (eNOS) was postulated to be associated with hyperhomocysteinemia and could influence individual susceptibility to CAD. The aims of this study were to investigate (a) the relationship of the eNOS G894T polymorphism with the presence and the severity of CAD and (b) the possible relationship between hyperhomocysteinemia and the eNOS G894T variant for the risk of CAD severity in a Tunisian population. METHODS: We used PCR with restriction fragment length polymorphism analysis to detect the G894T variant of the eNOS gene in 100 patients with CAD and 120 healthy controls. The severity of CAD was expressed by the number of affected vessels. Total plasma homocysteine concentrations were determined by direct chemiluminescence assay. RESULTS: The frequencies of the eNOS GG, GT, and TT genotypes in the CAD group were significantly different from those in the control group (45%, 44%, and 11% vs 60%, 35.8% and 4.2%, respectively; P = 0.035). There was no association between the eNOS G894T genotype frequencies and the number of stenosed vessels (P = 0.149). In the CAD group, the coexistence of the 894 GT or TT genotypes and hyperhomocysteinemia led to an increased risk of CAD severity. CONCLUSION: The G894T polymorphism of the eNOS gene is associated with the presence of CAD, and in conjunction with hyperhomocysteinemia, increased the risk of CAD severity in a Tunisian population.     

吸烟者内皮型一氧化氮合酶基因G894T多态性与冠心病的相关性研究

目的探讨吸烟者内皮型一氧化氮合酶基因（eNOS）G894T多态性与冠状动脉粥样硬化性心脏病（冠心病）的发生及其严重程度间的关系。方法将203例吸烟者和182例非吸烟者根据冠状动脉造影结果分为冠心病组（196例）和对照组（189例）,以冠状动脉病变支数判定严重程度。以聚合酶链式反应-限制性片段长度多态性（PCR—RFIP）检测eNOS基因G894T多态性,按吸烟与否分析G894T多态性与冠心病的关系。结果冠心病组GT＋TT基因型分布与T基因突变频数明显高于对照组（x^2=4．26、6．21,P均〈0．05）;吸烟者基因GT＋TT型及T等位基因频率均显著高于非吸烟者（x^2〈5．82、5．77,P均〈0．05）;冠心病组eNOS基因型分布在单支与多支病变组之间差异无统计学意义（x^2=3．36,P〉0．05）,而吸烟的冠心病患者差异有统计学意义（x^2=6.48,P〈0．05）,吸烟的GG＋TT型者更可能发生多支病变,OR=3．42,95％CI（1．440—4.400）。结论eNOS894位点G→T、吸烟与冠心病的发生及其严重程度间的关系密切。     

Smoking status-dependent association of the 27-bp repeat polymorphism in intron 4 of endothelial nitric oxide synthase gene with plasma nitric oxide concentrations

BACKGROUND: Euonymus alatus (EA) has been used for tumor therapy. However, it is still unclear how this herb prevents the diseases in experimental models. Nitric oxide (NO) as a potent macrophage-derived effector molecule against a variety of tumors has received increasing attention. METHODS: Using mouse peritoneal macrophages, we have examined the mechanism by which EA regulates NO production. RESULTS: When EA was used in combination with recombinant interferon-gamma (rIFN-gamma), there was a marked cooperative induction of NO production. However, EA had no effect on NO production by itself. The increased production of NO from rIFN-gamma plus EA-stimulated cells was almost completely inhibited by pre-treatment with pyrrolidine dithiocarbamate (PDTC), an inhibitor of nuclear factor kappa B (NF-kappaB). Furthermore, treatment of peritoneal macrophages with rIFN-gamma plus EA caused a significant increase in tumor necrosis factor-alpha (TNF-alpha) production. PDTC also decreased the effects of EA on TNF-alpha production significantly. CONCLUSIONS: EA increases the production of NO and TNF-alpha by rIFN-gamma-primed macrophages and suggest that NF-kappaB plays a critical role in mediating these effects of EA.     

Impact of endothelial nitric oxide synthase gene polymorphism on severity of enterovirus 71-infection in Chinese children

Objectives

Genetic polymorphism G894T on the endothelial nitric oxide synthase (eNOS) gene has been reported as a susceptibility factor in a number of diseases, but evidence of its effect on enterovirus 71 (EV71) infection is lacking. This study investigated the possible association between this polymorphism (rs1799983) and disease severity in Chinese children with EV71 infection.

Design and methods

185 children with EV71 infection (83 with severe and 102 with mild disease) and 234 control healthy children underwent testing with polymerase chain reaction–restriction fragment length polymorphism (PCR–RLFP) to detect G894T polymorphism. In addition, plasma levels of nitric oxide (NO), interleukin 1 beta (IL-1β), interleukin 6 (IL-6), and tumor necrosis factor-alpha (TNF-α) and serum eNOS activity were measured according to genotype.

Results

The presence of GT + TT genotypes and T allele were associated with severe cases compared to genotype GG (OR 2.5, 95% CI 1.2–5.3, P = 0.017) and G (OR 2.4, 95% CI 1.2–4.8, P = 0.011). Furthermore, in EV71 encephalitis, GT + TT genotype and T allele were also more frequent than GG and G (P < 0.05). The NO level and eNOS activity in T carriers (GT + TT) (84.3 ± 2.5 μmol/L and 14.4 ± 1.8 U/mL) were significantly less compared to in G carriers (GG) (92.0 ± 1.5 μmol/L and 19.1 ± 1.7 U/mL, P < 0.001). But T carriers had higher plasma levels of IL-1β, IL-6, and TNF-α than people without a T allele (P < 0.001), and a significant negative correlation was observed between NO and cytokine levels.

Conclusion

The results indicate that carrying the T allele of the eNOS G894T gene polymorphism was associated with EV71 infection, and could be a susceptibility factor in the development of EV71 infection in Chinese children.     

-786T>C polymorphism of the endothelial nitric oxide synthase gene in Chilean subjects with coronary artery disease and controls

BACKGROUND: The vascular endothelium plays an important role in the atherosclerotic process through the release of mediators such as nitric oxide. The NO relaxes vascular smooth muscle, inhibits platelet activation, and modulates migration and growth of vascular smooth muscle cells; consequently eNOS may have a significant atheroprotective function through this mechanism. The -786T>C polymorphism of the eNOS gene has been implicated in the development of coronary artery disease (CAD). We investigated the possible association between presence of CAD documented by angiography and the -786T>C polymorphism of the NOS3 gene in Chilean individuals. METHODS: A total of 112 unrelated patients with diagnosis of CAD and 109 controls were included in this study. The -786T>C gene polymorphism was analyzed by PCR-RFLP. RESULTS: The frequency of CC homozygous genotype for -786T>C polymorphism was 6% in CAD patients and 4% in the control group. However, the genotype distribution and allele frequencies were not significantly different between CAD and control subjects (P=NS). Moreover, the odds ratio for CAD associated with the C variant failed to reach statistical significance (OR=1.03; 95% CI: 0.60-1.76, P=NS). CONCLUSION: These findings suggest that the -786T>C polymorphism of the eNOS gene was not associated with CAD in study Chilean individuals.