Background-Coronary spasm plays an important role in the pathogenesis of ischemic heart diseases in general. However, the precise mechanism(s) responsible for coronary spasm remains to be elucidated, and we examined the molecular genetics of coronary spasm. Methods and Results-We searched for the possible mutations in the endothelial nitric oxide synthase (eNOS) gene in patients with coronary spasm. In this study, we demonstrate the existence of 3 linked mutations in the 5Ј-flanking region of the eNOS gene (T

clinicaltrials.gov Identifier: NCT00110448.

Abstract-Essential hypertension has a genetic basis. Accumulating evidence, including findings of elevation of arterial blood pressure in mice lacking the endothelial nitric oxide synthase (eNOS) gene, strongly suggests that alteration in NO metabolism is implicated in hypertension. There are, however, no reports indicating that polymorphism in the eNOS gene is associated with essential hypertension. We have identified a missense variant, Glu298Asp, in exon 7 of the eNOS gene and demonstrated that it is associated with both coronary spastic angina and myocardial infarction. To explore the genetic involvement of the eNOS gene in essential hypertension, we examined the possible association between essential hypertension and several polymorphisms including the Glu298Asp variant, variable number tandem repeats in intron 4 (eNOS4b/4a), and two polymorphisms in introns 18 and 23. We performed a large-scale study of genetic association using two independent populations from Kyoto (nϭ458; 240 normotensive versus 218 hypertensive subjects) and Kumamoto (nϭ421; 223 normotensive versus 187 hypertensive subjects), Japan. In both groups, a new coding variant, Glu298Asp, showed a strong association with essential hypertension (Kyoto: odds ratio, 2.3 [95% confidence interval, 1.4 to 3.9]; Kumamoto: odds ratio, 2.4 [95% confidence interval, 1.4 to 4.0]). The allele frequencies of 298Asp in hypertensive subjects were significantly higher than those in normotensive subjects in both groups (Kyoto: 0.103 versus 0.050, PϽ0.0017; Kumamoto: 0.120 versus 0.058, PϽ0.0013, respectively). No such disequilibrium between genotypes was significantly associated with any other polymorphisms we examined; the Glu298Asp variant was also not linked to any other polymorphisms. In conclusion, the Glu298Asp missense variant was significantly associated with essential hypertension, which suggests that it is a genetic susceptibility factor for essential hypertension.(Hypertension. 1998;32:3-8.)Key Words: genes Ⅲ nitric oxide synthase Ⅲ hypertension, essential Ⅲ polymorphism Ⅲ genetics W ith a genetic contribution of from 25% to 60%, human essential hypertension has a genetic basis. Among persons younger than age 50 years, essential hypertension occurs 3.8 times more often in those having two or more first-degree relatives who developed high blood pressure before age 55.1 NO synthesis by the vascular endothelium is important for the regulation of vasodilator tone and the control of blood pressure in humans.2 A recent study using mice with disrupted eNOS gene revealed that eNOS function is required for vascular and hemodynamic responses to acetylcholine and that the disruption of the eNOS gene leads to hypertension. 3 Moreover, recent reports demonstrate that whole-body NO production in patients with essential hypertension is diminished under basal conditions, as established by measurement of urinary and plasma nitrate. 4 In addition, the offspring of hypertensive patients exhibit a reduced response to acetylcholine linked to a defect in the NO pathway.5...

Coronary spasm plays an important role in the pathogenesis of not only variant angina but also ischemic heart disease in general. However, the precise mechanism(s) by which coronary spasm occurs remains to be elucidated. Coronary spasm may arise from interactions between environmental and genetic factors. Endothelial-derived nitric oxide (NO) has been implicated in the control of vascular tone. We have recently shown that both basal and acetylcholine (ACh)-induced NO activities are impaired in the coronary arteries of patients with coronary spasm. The purpose of this study has been to elucidate the possible variants that occur in the coding region of the endothelial nitric oxide synthase (eNOS) gene and that may be associated with coronary spasm. After initial screening in the entire 26 coding regions of the eNOS gene, we found a missense Glu298Asp variant in exon 7 in patients with coronary spasm. We subsequently performed a larger scale study involving 113 patients with coronary spasm and 100 control subjects, who were all diagnosed by intracoronary injection of ACh. The analysis revealed a significant difference in the distribution of the variant between the coronary spasm group (21.2%) and control group (9.0%; P=0.014 for dominant effect). Thus, we have found the missense Glu298Asp variant in the eNOS gene by the analysis of its entire 26 coding regions. The variant is significantly associated with coronary spasm.

Background-Recent reports have indicated that aldosterone is produced in extra-adrenal tissues in animals. The present study was designed to examine whether aldosterone is produced in human heart. Methods and Results-Plasma levels of aldosterone, BNP, and angiotensin-converting enzyme were measured in anterior interventricular vein (AIV), coronary sinus (CS), and aortic root (Ao), respectively, in 20 patients with left ventricular systolic dysfunction (LVSD), 25 patients with LV diastolic dysfunction (LVDD), and 23 control subjects. Aldosterone levels were significantly higher in AIV and CS than Ao in LVSD (98Ϯ10 versus 72Ϯ9 pg/mL, PϽ0.001, and 97Ϯ11 versus 72Ϯ9 pg/mL, PϽ0.001, respectively) and LVDD (87Ϯ10 versus 71Ϯ9 pg/mL, PϽ0.01, and 84Ϯ10 versus 71Ϯ9 pg/mL, PϽ0.01, respectively) groups, but no differences were observed in levels for these sites in the control group. Levels of ACE activity and BNP also were higher in AIV than Ao in both LV dysfunction groups. The difference in aldosterone levels between AIV and Ao and those in BNP and angiotensin-converting enzyme had a significant positive correlation with LVEDP and a significant negative correlation with LV ejection fraction in the LVSD group. Conclusions-Production

There was a significant association of the missense Glu298Asp variant of the eNOS gene with MI. This marker-disease association may be due to the impaired effects of NO on the cardiovascular system: dysregulation of vascular tone, platelet aggregation and leukocyte adhesion and smooth muscle cell proliferation, all of which promote coronary atherosclerosis and thrombosis.

We recently reported that a mutation (-786T-->C) in the promoter region of the endothelial nitric oxide synthase (eNOS) gene reduced transcription of the gene and was strongly associated with coronary spastic angina and myocardial infarction. To elucidate the molecular mechanism for the reduced eNOS gene transcription, we have now purified a protein that specifically binds to the mutant allele in nuclear extracts from HeLa cells. The purified protein was identical to replication protein A1 (RPA1), known as a single-stranded DNA binding protein essential for DNA repair, replication and recombination. In human umbilical vein endothelial cells, inhibition of RPA1 expression using antisense oligonucleotide restored transcription driven by the mutated promoter sequence, whereas, conversely, overexpression of RPA1 further reduced it. RPA1 was similarly detected in placenta and eNOS mRNA levels in placentas carrying the -786T-->C mutation were significantly lower than in placentas without it. The functional importance of the diminished eNOS expression was revealed by the finding that serum nitrite/nitrate levels among individuals carrying the -786T-->C mutation were significantly lower than among those without the mutation. RPA1 thus apparently functions as a repressor protein in the -786T-->C mutation-related reduction of eNOS gene transcription associated with the development of coronary artery disease.

Background-Cannabinoid 1 (CB1) receptor blockade with rimonabant represents a clinical therapeutic strategy for obesity. Recently, the role of the endocannabinoid system has been described in peripheral organs. We sought to determine whether the endocannabinoid system could be involved in human atherosclerosis and whether CB1 receptor blockade could modulate proinflammatory activity in macrophages. Methods and Results-mRNA expression levels of CB1 receptor in coronary atherectomy samples were significantly higher in patients with unstable angina than in those with stable angina (3.62Ϯ2.96-fold; nϭ7; PϽ0.05). Immunoreactive area analysis of the coronary artery showed that CB1 receptor expression was greater in lipid-rich atheromatous plaques than in fibrous plaques, especially in CD68 macrophages (9.5Ϯ1.2% versus 0.6Ϯ0.6%; nϭ5; PϽ0.01). Levels of blood endocannabinoids were significantly higher in patients with coronary artery disease (nϭ20) than those without coronary artery disease (nϭ20) In cultured macrophages, expression of CB1 receptor was significantly increased during monocyte-macrophage differentiation (1.78Ϯ0.13-fold; nϭ6; PϽ0.01). CB1 receptor blockade in macrophages induced a significant increase in cytosolic cAMP (29.9Ϯ13.0%; nϭ4; PϽ0.01), inhibited phosphorylation of c-Jun N-terminal kinase (Ϫ19.1Ϯ12.6%, nϭ4; PϽ0.05), and resulted in a significant decrease in the production of proinflammatory mediators (interleukin-1␤, Ϫ28.9Ϯ10.9%; interleukin-6, Ϫ24.8Ϯ7.6%; interleukin-8, Ϫ22.7Ϯ5.2%; tumor necrosis factor-␣, Ϫ13.6Ϯ4.8%; matrix metalloproteinase-9, Ϫ16.4Ϯ3.8%; nϭ4 to 8; PϽ0.01). Conclusions-Patients with coronary artery disease demonstrated the activation of the endocannabinoid system with elevated levels of blood endocannabinoids and increased expression of CB1 receptor in coronary atheroma. CB1 receptor blockade exhibited antiinflammatory effects on macrophages, which might provide beneficial effects on atherogenesis. (Circulation. 2009;119:28-36.)Key Words: atherosclerosis Ⅲ inflammation Ⅲ macrophages Ⅲ obesity Ⅲ receptors, cannabinoid O verweight and obesity (particularly visceral obesity) are quickly reaching global epidemic proportions and are associated with the metabolic syndrome, correlating with an increased risk of cardiovascular morbidity and mortality. [1][2][3] However, lifestyle intervention programs aimed at reducing the risk of cardiovascular disease in obese patients do not appear to provide long-lasting success. 4 Consequently, antiobesity pharmacotherapy has been recognized as an important adjunctive therapy to lifestyle modification to prevent cardiovascular complications. Clinical Perspective p 36The recently discovered endocannabinoid system contributes to the physiological regulation of energy balance, food intake, and metabolism of lipids and glucose. 5 This system consists mainly of 2 endogenous ligands, anandamide and 2-arachidonoyl glycerol (2-AG), and 2 types of G protein-coupled cannabinoid receptors, namely cannabinoid 1 (CB1) receptor and cannabinoid 2 (CB2) receptor...