N. Correa

Deregulation of Soluble Adhesion Molecules in Resistant Hypertension and Its Role in Cardiovascular Remodeling

BACKGROUND Resistant hypertension (RHTN) and target organ damage are linked to increased inflammatory biomarkers, which may regulate adhesion molecules, such as intracellular adhesion molecule-1 (ICAM-1); vascular cell adhesion molecule-1 (VCAM-1); and the platelet (P-selectin) and endothelial (E-selectin) selectins. We investigated a previously unknown relationship between soluble P-selectin (sP-selectin), E-selectin (sE-selectin), ICAM-1 (sICAM-1) and VCAM-1 (sVCAM-1) with RHTN and target organ damage. METHODSANDRESULTS We included 110 subjects diagnosed for true RHTN and 112 mild-moderate hypertensive (HTN) patients. Blood pressure parameters, pulse wave velocity and left ventricular mass index (LVMI) were measured. Adhesion molecules were measured on ELISA. Both sP-selectin and sE-selectin were increased; in contrast, sICAM-1 was reduced in RHTN compared with HTN patients, while similar sVCAM-1 was noted in the groups. sP-selectin and sVCAM-1 were elevated in the presence of arterial stiffness (sP-selectin: 104±47 vs. 89±45 ng/ml, P<0.05; sVCAM-1: 1,189±411 vs. 1,060±412 ng/ml, P<0.05) and cardiac hypertrophy (sP-selectin: 105±51 vs. 88±43 ng/ml, P<0.05; sVCAM-1: 1,170±433 vs. 1,040±383 ng/ml, P<0.05) in all HTN patients. sP-selectin was associated with target organ damage after adjustment for age and BP. Apart from potential confounders, sE-selectin was a significant indicator of RHTN. CONCLUSIONS The adhesion molecule sP-selectin plays a role in cardiovascular damage, and sE-selectin in resistance to antihypertensive therapy. (Circ J 2016; 80: 1196-1201).

Adiponectin -11377C/G and +276G/T polymorphisms affect adiponectin levels but do not modify responsiveness to therapy in resistant hypertension.

Resistant hypertension (RHTN) is a multifactorial and polygenic disease, frequently associated with obesity. Low plasma adiponectin levels, a hormone produced by the adipose tissue, were associated with RHTN. Single nucleotide polymorphisms (SNPs) ‐11377C/G (rs266729) and +276G/T (rs1501299) in ADIPOQ (adiponectin gene) were associated with hypertension. This study evaluated the association between two SNPs (‐11377C/G and +276G/T) and adiponectin levels in RHTN. This study comprised 109 patients with RHTN genotyped for both polymorphisms. A cross‐sectional study was designed to compare features of CC homozygous versus G allele carriers for ‐11377C/G and GG homozygous versus T allele carriers for +276G/T. Office and ambulatory BP measurements were similar among genotypes subgroups in both SNPs as well as the markers of target organ damage (arterial stiffness, left ventricular mass index and microalbuminuria). Adiponectin concentrations were significantly higher in CC compared to G carrier for ‐11377C/G (CC:7.0 (4.0–10.2) versus G allele:5.5 (2.5–7.9), p = 0.04) and lower in GG compared to T carrier for +276G/T (GG:5.3 (2.3–7.7) versus T allele:7.1 (3.6–10.5), p = 0.04). Adjusting for systolic ambulatory BP, body mass index, age, gender, race and presence of type 2 diabetes, multiple linear regression analyses revealed that the minor alleles G (β‐coefficient= ‐0.14, SE=0.07, p = 0.03) and T (β‐coefficient=0.12, SE=0.06, p = 0.04) were independent predictors of adiponectin. The ‐11377C/G and +276G/T SNPs in ADIPOQ were associated with adiponectin levels in RHTN individuals.

Matrix metalloproteinase-2 − 735C/T polymorphism is associated with resistant hypertension in a specialized outpatient clinic in Brazil

BACKGROUND Matrix metalloproteinases (MMPs) are enzymes involved in cardiovascular (CV) remodeling and hypertension-mediated target organ damage (TOD). Genetic polymorphisms in matrix metalloproteinase 2 (MMP-2) gene [-1575G/A (rs243866); -1306C/T (rs243865); and -735C/T (rs2285053)] are associated with several CV conditions, however the relationship between MMP-2 polymorphisms and resistant hypertension (RH) is unknown. We evaluated whether these genetic single nucleotide polymorphisms (SNPs) in MMP-2 gene are associated with 1) MMP-2 and tissue inhibitor of metalloproteinase-2 (TIMP-2) levels in RH and mild to moderate hypertensive (HT) subjects, 2) left ventricular hypertrophy (LVH) and arterial stiffness and 3) the presence of RH. METHODS One hundred and nineteen RH and 136 HT subjects were included in this cross-sectional study. Genotypes were determined by real-time PCR using TaqMan probes. Haplotypes were estimated using Bayesian method. RESULTS The levels of MMP-2 and TIMP-2 were similar among genotypes and haplotypes for the three studied polymorphisms in HT and RH groups. RH showed higher frequency for GCC haplotype and lower frequency of GCT and ATC haplotypes (-1575G/A, -1306C/T and -735C/T, respectively) compared to HT (0.77 vs. 0.64; 0.09 vs. 0.17; 0.13 vs. 0.19, p=0.003 respectively). GCC haplotype was associated to RH apart from potential confounders (odds ratio (OR)=2.09; 95% confidence interval (CI)=1.20-3.64; p=0.01). In addition, CC genotype (OR=2.93; 95% CI=1.22-7.01; p=0.02) and C allele (OR=2.81; 95% CI=1.26-6.31; p=0.01) for -735C/T polymorphism were independently associated with RH. GCT haplotype was associated with reduced probability of having RH (OR=0.35; 95% CI=0.16-0.79; p=0.01). Finally, no relationship was found between studied MMP-2 SNPs and left ventricular hypertrophy and arterial stiffness in both groups. CONCLUSION GCC haplotype carriers showed higher probability to have RH (odds ratio>1), while the GCT haplotype carriers showed lower probability to have RH, suggesting that the GCT haplotype may represent a protective genetic factor for the development of RH. These finds suggest that GCC and GCT haplotypes, and C allele and CC genotype of the -735C/T MMP-2 gene polymorphism may have a role in RH.

Crosstalk between obesity and MMP-9 in cardiac remodelling –a cross-sectional study in apparent treatment-resistant hypertension

Abstract The balance between matrix metalloproteinases (MMP) and their tissue inhibitors (TIMP) plays a key role in the development of hypertension and obesity. We aimed to evaluate the levels of MMP-2 and 9 and TIMP-2 and -1 in obese and non-obese apparent treatment-resistant hypertensive subjects (aTRH) and its association with cardiac hypertrophy. This cross-sectional study enrolled 122 subjects and divided into obese aTRH (n = 67) and non-obese (n = 55) group. Clinical and biochemical data were compared between both groups, including office BP, ambulatory BP, plasma MMP-2 and 9, TIMP-2 and 1 and left ventricular mass index (LVMI). We found higher MMP-9 levels and MMP-9/TIMP-1 ratio in obese aTRH subjects but no difference in MMP-2 and TIMP-1 levels. Obesity influenced MMP-9 levels [β = 20.8 SE =8.6, p = 0.02) independently of potential confounders. In addition, we found a positive correlation between MMP-9 and anthropomorphic parameters. Finally, obese aTRH subjects with left ventricular hypertrophy (LVH) had greater MMP-9 levels compared with non-obese with LVH. Our study suggests that MMP-9 levels are influenced by obesity and may directly participate in the progressive LV remodelling process, suggesting a possible role for a higher cardiovascular risk in apparent resistant hypertensive subjects.

The rs243866/243865 polymorphisms in MMP-2 gene and the relationship with BP control in obese resistant hypertensive subjects

We sought to investigate whether the polymorphisms rs243865 (-1306C>T); rs243866 (-1575G>A) and rs2285053 (-735C>T) in metalloproteinases 2 - MMP-2 gene and rs17576 (Q279R), rs17577 (Q668R) and rs3918242 (-1562C>T) in MMP-9 gene are associated with clinical outcomes in obese resistant hypertensive (RH) subjects. One hundred and twenty RH were enrolled in this cross-sectional study and divided into obese (n=63) and non-obese (n=57) according to body mass index. Genotypes were determined by real-time PCR using TaqMan probes. We determined pulse wave velocity (PWV), microalbuminuria and left ventricular mass index (LVMI) to assess TODs. Obese and non-obese RH had similar allele, genotype and haplotype distributions for all polymorphisms assessed but obese RH subjects carrying the low frequency allele for SNPs in MMP-2 gene had higher ambulatory diastolic blood pressure. Also, PWV and LVMI were higher in subjects carrying the low frequency allele for SNPs in MMP-2 gene. Regarding MMP-9 gene, office diastolic BP levels were higher in the AA genotype individuals compared to the G allele group for rs17576 polymorphism, while the opposite was found regarding the microalbuminuria level. Independent multiple linear regression analyses revealed that both A allele for rs243865 and T allele for rs243866 in MMP-2 gene were associated with ambulatory diastolic levels in obese RH subjects, apart from potential confounders. Our study suggests that rs243866/rs243865 in the MMP-2 gene are related to BP levels in obese RH subjects, although TODs present in this population seem to be dependent of a combination of other factors besides the genetic polymorphisms.

[PP.25.06] ACUTE EFFECTS OF AEROBIC AND RESISTANCE EXERCISES IN INFLAMMATORY MARKERS IL-10 AND IL-1RA IN PATIENTS WITH RESISTANT HYPERTENSION: RATIONALE OF A RANDOMIZED, CONTROLLED TRIAL

Objective: The role of regular physical activity on blood pressure (BP) and inflammatory process have been investigated in hypertension. This study aims to evaluate the acute effects of either aerobic, resistance or both combined exercises on inflammatory markers IL-10 and IL-1ra in resistant (RH) and controlled hypertensive (HT) subjects. Secondly, we will assess those effects on BP levels. Design and method: This randomized, single-blind, crossover non-pharmacological intervention study will include 20 patients, 10 RH and 10 HT, who are regularly followed at the Outpatient Resistant Hypertension Clinic (UNICAMP, Brazil). This study will comprise two parts: (1) to determine the type and duration of each exercise individually for each volunteers, and (2) to perform all the three types of exercise in a crossover way in all subjects, which will consist of: 1) aerobic exercise: activity on a treadmill lasting 45 minutes with intensity of 50–60% of maximum heart rate obtained from ergometer test; 2) resistance exercise: 4 series of 12 repetitions of resistance exercises at moderate intensity (until moderate fatigue), for 45 minutes; 3) combined exercise: aerobic (25 minutes) + resistance (20 minutes), with an interval of 2 minutes between sessions totalizing 45 minutes. We will perform blood test before and after the interventions to assess inflammatory biomarkers IL-10 and IL-1ra. Also, we will determine the beat-to-beat BP levels (using the Finometer device). Figure. No caption available. Results: We expect that the acute practice of exercise modulates the inflammatory biomarkers, IL-10 and IL-1ra, and secondly, the BP levels. Conclusions: This study may provide a better understanding of the acute mechanisms of this non-pharmacologic treatment in RH, in order to make this more effective in this high-risk population.

[BP.02.06] MATRIX METALLOPROTEINASE 2 AND 9 POLYMORPHISMS ARE ASSOCIATED WITH TARGET ORGAN DAMAGE IN OBESE RESISTANT HYPERTENSIVE SUBJECTS

Objective: The aim of this study is to analyze the influence of MMP-2 and -9 SNPs in obese hypertensive (HTN) and resistant hypertensive (RH) subjects, as well as their association with MMP-2 and 9 levels and target organ damage (TOD). Design and method: Two thousand and fifty six hypertensive subjects were divided in obese [body mass index (BMI higher than 30 kg/m2) and non-obese (BMI lower than 30 kg/m2). Genotypes were obtained by allelic discrimination assay using real time polymerase chain reaction. We compare clinical and laboratorial characteristics according to genotypes/haplotypes for MMP-2 (rs243865, rs243866 and rs2285053) and for MMP-9 SNPs (rs17577, rs17576 and rs391824) in obese HTN and RH subjects. Results: No difference in allele, genotype and haplotype frequencies for all polymorphisms between obese and non-obese HTN and RH were found. MMP-2, MMP-9 and their TIMPs levels were similar in obese HTN and RH according to MMP-2 and MMP-9 genotypes for all SNPs assessed. The same happened for clinical and biochemical characteristics among the genotypes in obese HTN and RH subjects except for some parameters: diastolic ambulatory blood pressure (BP) monitoring was higher in AG+AA compared to GG genotype for both rs243866 and rs243865 MMP-2 SNPs and the office diastolic BP was higher in AA than AG+GG genotype for rs17576 MMP-9 polymorphism in obese RH. However, when we compared the TOD according to genotype in obese RH we found that PWV was higher in CC genotype than CT for rs2285053 MMP-2 polymorphism [8.8 (8 – 11) vs 7.8 (6 – 8), p = 0.04]. Also, the AG+AA genotype for rs243866 and rs243865 MMP-2 polymorphisms have the same levels of LVMI and they are higher than GG genotype [116 ± 37 vs 138 ± 40, p = 0.04]. Finally, the microalbuminuria level was higher in AG+GG compared to AA genotype for rs17576 MMP-9 polymorphism. A multiple linear regression showed that only rs243866 and rs243865 are an independent predictor for LVMI levels after adjusted by gender, age, office BP, aldosterone and glucose levels. Conclusions: Therefore, the MMP-2 and -9 polymorphisms are associated with TOD in obese RH subjects.

[PP.25.16] THE INFLAMMATORY STATUS SCORE INCLUDING TNF-ALPHA, IL-6, IL-8, IL-10, LEPTIN AND ADIPONECTIN IS ASSOCIATED WITH RESISTANT HYPERTENSION

Objective: Subclinical systemic inflammation has been indicated to be present in resistant hypertension (RHTN). The aim of the study was to develop an integrated measure of the circulating cytokines/adipokines that underlie the pathophysiology of RHTN. Design and method: RHTN (n = 112) and mild to moderate hypertensive (HTN) subjects (n = 112) were studied in a cross-sectional design. Plasma cytokine/adipokine [TNF-alpha, IL-6, IL-8, IL-10, leptin and adiponectin] values were divided into tertiles. A score ranging from 1 (lowest tertile) to 3 (highest tertile) was assigned. The inflammatory score (IS) was the sum of each pro-inflammatory cytokine score from which adiponectin and IL-10 – anti-inflammatory cytokines – scores were subtracted in each study subject. Results: IS was higher in RHTN subjects compared to their counterparts (4.2 ± 2.2 vs. 3.5 ± 1.9; p = 0.02). IS positively correlated with obesity parameters, such as body mass index (r = 0.40; p < 0.001), waist circumference (r = 0.30; p < 0.001) and fat mass assessed by bioimpedance (r = 0.31; p < 0.001) in all subjects. Logistic regression analyses revealed that IS was an independent predictor of RHTN (OR = 1.19; p = 0.02), apart from age, gender and race, although it did not remain significant after BMI adjustment. Conclusions: A state of subclinical inflammation defined by IS including TNF-alpha, IL-6, IL-8, IL-10, leptin and adiponectin is associated with RHTN. In addition, this score strongly correlates with obesity parameters, independently of hypertensive status. This integrated measure may represent a new tool for the evaluation of the severity of low-grade inflammation in order to identify RHTN patients; also highlights the contribution of obesity to inflammatory process.

MCP-1 Levels are Associated with Cardiac Remodeling but not with Resistant Hypertension

Background Hypertension is a chronic, low-grade inflammation process associated with the release of cytokines and development of target organ damage. Deregulated monocyte chemoattractant protein-1 (MCP-1) levels have been associated with high blood pressure and cardiovascular complications; however, the mechanisms involved are complex and not fully understood. Objective This study aimed to compare the levels of MCP-1 in patients with resistant (RH) versus mild-to-moderate (HTN) hypertension and their association with the presence or absence of left ventricular hypertrophy (LVH) in all hypertensive subjects. Methods We enrolled 256 hypertensive subjects: 120 RH and 136 HTN, investigating the relationship between circulating MCP-1 levels and blood pressure, biochemical data, hematologic profile, and cardiac damage within the RH and HTN groups. Plasma MCP-1 levels were measured by ELISA and LVH was assessed by echocardiography. Results We found no difference in MCP-1 levels between RH and HTN subjects. On the other hand, we encountered lower MCP-1 levels in patients with LVH (105 pg/mL [100 - 260 pg/mL] versus 136 pg/mL (100 - 200 pg/mL), p = 0.005, respectively] compared with those without LVH. A logistic regression model adjusted for body mass index (BMI), age, race, aldosterone levels, and presence of diabetes and RH demonstrated that median levels of MCP-1 (2.55 pg/mL [1.22 - 5.2 pg/mL], p = 0.01) were independently associated with LVH in the entire hypertensive population. Conclusion Since MCP-1 levels were similar in both RH and HTN subjects and decreased in hypertensive patients with existing LVH, our study suggests a possible downregulation in MCP-1 levels in hypertensive individuals with LVH, regardless of hypertension strata.

[PP.23.07] ACUTE SILDENAFIL USE LOWER 24H BLOOD PRESSURE LEVELS IN RESISTANT HYPERTENSION A RANDOMIZED, PLACEBO CONTROLLED, CROSS-OVER TRIAL

Objective: The present study sought to evaluate whether acute administration of sildenafil compared with placebo improves ambulatory BP levels in RHTN subjects. Design and method: This interventional, single-blinded, placebo-controlled, one-way crossover trial included 26 patients with true RHTN. Increasing oral doses of sildenafil were given at 30 minute-interval (37.5 mg, 50 mg and 100 mg) in a single day. After a washout period of 14 days, patients received consecutive oral doses of placebo and the protocol was repeated. Before and after each protocol day (sildenafil and placebo), patients underwent 24-hour ABPM evaluation. Results: The reduction of systolic (−8.8 ± 1.4 vs. 1.3 ± 1.2mmHg), diastolic (−5.3 ± 3.3 vs. 1.8 ± 1.1mmHg) and mean (−7.9 ± 3.6 vs. 0.8 ± 0.9mmHg) 24-hour BP were higher after sildenafil compared with placebo. The main differences were observed on daytime BP levels (systolic; −6 ± 4.7 vs. 4.4 ± 1.5 mmHg; and mean: −4.8 ± 3.9 vs. 3.5 ± 1.4 mmHg; sildenafil vs. placebo, respectively). Conclusions: Our study suggests that an acute high-dose load of sildenafil improves ABPM parameters in resistant hypertensive patients. Considering its antihypertensive effect, sildenafil may represent a therapeutic option for the treatment of RHTN.