Jackeline S. Rangel Machado

Matrix metalloproteinase-9 polymorphisms affect plasma MMP-9 levels and antihypertensive therapy responsiveness in hypertensive disorders of pregnancy.

Abnormal matrix metalloproteinase (MMP)-9 levels may have a role in hypertensive disorders of pregnancy. We examined whether MMP-9 genetic polymorphisms (g.−1562C>T and g.−90(CA)13−25) modify plasma MMP-9 and tissue inhibitor of metalloproteinase (TIMP)-1 levels and the responses to antihypertensive therapy in 214 patients with preeclampsia (PE), 185 patients with gestational hypertension (GH) and a control group of 214 healthy pregnant (HP). Alleles for the g.−90(CA)13−25 polymorphism were grouped L (low) (<21 CA repeats) or H (high) (⩾21 CA repeats). Plasma MMP-9 and TIMP-1 concentrations were measured by enzyme-linked immunosorbent assay. Plasma MMP-9 concentrations were not affected by genotypes or haplotypes in HP and PE groups, except for the g.−90(CA)13−25 polymorphism: GH patients with the LH genotype for this polymorphism have higher MMP-9 levels than those with other genotypes. The T allele for the g.−1562C>T polymorphism and the H4 haplotype (combining T and H alleles) are associated with GH and lack of responsiveness to antihypertensive therapy in GH. The H2 haplotype (combining C and H alleles) was associated with lack of responsiveness to antihypertensive therapy in PE, but not in GH. In conclusion, our results show that MMP-9 genetic variants are associated with GH and suggest that MMP-9 haplotypes affect the responsiveness to antihypertensive therapy in hypertensive disorders of pregnancy.

Association between matrix metalloproteinase (MMP)-2 polymorphisms and MMP-2 levels in hypertensive disorders of pregnancy

We examined whether two functional polymorphisms (g.-1306C>T and g.-735C>T) in matrix metalloproteinase (MMP)-2 gene are associated with preeclampsia (PE) or gestational hypertension (GH), and whether they modify MMP-2 or tissue inhibitor of metalloproteinase (TIMP)-2 plasma concentrations in these hypertensive disorders of pregnancy. We studied 130 healthy pregnant (HP), 130 pregnant with GH, and 133 pregnant with PE. Genomic DNA was extracted from whole blood and genotypes for g.-1306C>T and g.-735C>T polymorphisms were determined by Real Time-PCR, using Taqman allele discrimination assays. Haplotypes were inferred using the PHASE program. Plasma MMP-2 and TIMP-2 concentrations were measured by ELISA. The main findings were that pregnant with PE have higher plasma MMP-2 and TIMP-2 concentrations than HP (P<0.05), although the MMP-2/TIMP-2 ratios were similar (P>0.05). Moreover, pregnant with GH have elevated plasma MMP-2 levels and MMP-2/TIMP-2 ratios compared to HP (P<0.05). While MMP-2 genotypes and haplotypes are not linked with hypertensive disorders of pregnancy, MMP-2 genotypes and haplotypes are associated with significant alterations in plasma MMP-2 and TIMP-2 concentrations in preeclampsia (P<0.05). Our findings may help to understand the relevance of MMP-2 and its genetic polymorphisms to the pathophysiology of hypertensive disorders of pregnancy. It is possible that patients with PE and the MMP-2 haplotype combining the C and T alleles for the g.-1306C>T and g.-735C>T polymorphisms may benefit from the use of MMPs inhibitors such as doxycycline. However, this possibility remains to be determined.

Positive correlations between circulating adiponectin and MMP2 in preeclampsia pregnant

OBJECTIVE The aims of the present study were to compare plasma concentrations of the adiponectin, leptin, metalloproteinases (MMP9 and MMP2) and its tissue inhibitors (TIMP1 and TIMP2) in preeclamptic (PE) and healthy pregnant (HP) groups and correlate them. METHODS A total of 105 pregnant women with pre-pregnancy body mass index (BMI) values ⩽ 30 kg/m(2) were enrolled for this study (59 PE and 46 HP). Biomarkers were measured using ELISAs. RESULTS Adiponectin (32%), leptin (45%), MMP2 (20%), TIMP1 (31%) and TIMP2 (23%) levels were higher in PE compared to HP (all P < 0.05). In addition there were positive correlations between adiponectin and MMP2 (r = 0.33; P = 0.03) and adiponectin and TIMP2 (r = 0.33; P = 0.03) in PE group, but not in HP. CONCLUSION Our findings show that adiponectin, leptin, MMP2, TIMP1 and TIMP2 levels are increased in PE and adiponectin may contribute to higher levels of MMP2 and TIMP2 in this disease.

Effects of Matrix Metalloproteinase (MMP)‐2 Polymorphisms on Responsiveness to Antihypertensive Therapy of Women with Hypertensive Disorders of Pregnancy

Imbalanced matrix metalloproteinase (MMP) expression, including MMP‐2, has been demonstrated in pre‐eclampsia. However, little is known about the effect of polymorphisms in MMP‐2 gene on hypertensive disorders of pregnancy. We examined whether two functional MMP‐2 polymorphisms (g.‐1306C>T and g.‐735C>T) are associated with pre‐eclampsia and/or gestational hypertension and whether these polymorphisms affect therapeutic responses in women with these conditions. We studied 216 healthy pregnant women (HP), 185 patients with gestational hypertension (GH) and 216 patients with pre‐eclampsia (PE). They were stratified as responsive or non‐responsive to antihypertensive therapy according to clinical and laboratorial parameters of therapeutic responsiveness. Genomic DNA was extracted from whole blood and genotypes for g‐1306C>T and g.‐735C>T polymorphisms were determined by real‐time PCR using Taqman allele discrimination assays. Haplotype frequencies were inferred using the PHASE 2.1 program. The distributions of MMP‐2 genotypes and haplotypes were similar in HP, GH and PE patients (p > 0.05). In addition, we found no significant differences in MMP‐2 genotype or haplotype frequencies when GH or PE patients were classified as responsive or non‐responsive to antihypertensive therapy (p > 0.05). Our results suggest that MMP‐2 polymorphisms do not affect the susceptibility to hypertensive disorders of pregnancy. In parallel, MMP‐2 polymorphisms apparently do not affect the responsiveness to antihypertensive therapy of women with these hypertensive disorders of pregnancy.

Tissue inhibitor of matrix metalloproteinase-1 polymorphism, plasma TIMP-1 levels, and antihypertensive therapy responsiveness in hypertensive disorders of pregnancy

Tissue inhibitor of metalloproteinase (TIMP)-1 is a major endogenous inhibitor of matrix metalloproteinase (MMP)-9, which may affect the responsiveness to therapy in hypertensive disorders of pregnancy. We examined whether TIMP-1 polymorphism (g.–9830T>G, rs2070584) modifies plasma MMP-9 and TIMP-1 levels and the response to antihypertensive therapy in 596 pregnant: 206 patients with preeclampsia (PE), 183 patients with gestational hypertension (GH) and 207 healthy pregnant controls. We also studied the TIMP-3 polymorphism (g.–1296T>C, rs9619311). Plasma MMP-9 and TIMP-1 levels were measured by ELISA. GH patients with the GG genotype for the TIMP-1 polymorphism had lower MMP-9 levels and MMP-9/TIMP-1 ratios than those with the TT genotype. PE patients with the TG genotype had higher TIMP-1 levels. The G allele and the GG genotype were associated with PE and responsiveness to antihypertensive therapy in PE, but not in GH. Our results suggest that the TIMP-1 g.–9830T>G polymorphism not only promotes PE but also decreases the responses to antihypertensive therapy.

Relationship between adiponectin and nitrite in healthy and preeclampsia pregnancies

BACKGROUND Controversial results have been reported regarding plasma adiponectin levels in preeclampsia (PE) compared to healthy pregnancies (HP). Adiponectin activates eNOS, increasing the levels of the vasodilator nitric oxide (NO). PE reduces the levels of nitrite (an NO marker) and induces higher levels of ADMA (an endogenous eNOS inhibitor) compared to HP. No previous study has examined whether a positive correlation exists between adiponectin and nitrite in HP and PE and how ADMA may interfere with this correlation. METHODS We measured plasma nitrite concentrations using an ozone-based chemiluminescence assay, and plasma ADMA and adiponectin levels using enzyme immunoassays in 117 pregnant women (70 healthy and 47 preeclamptic). RESULTS We found higher adiponectin levels (23.6±13.0 vs. 17.8±5.6µg/ml; P<0.05) and lower plasma nitrite levels (104.5±84.3 vs. 177.2±151.3 nM; P<0.05) in PE compared to HP. We found a significant positive correlation between these markers in HP (r=0.3; P<0.05), but no correlation in PE. However, when we grouped PE women regarding ADMA levels (low and high levels), a strong positive correlation was found in the group with lower ADMA levels (r=0.67; P<0.05), suggesting that high ADMA concentrations may interfere with the physiological activation of eNOS by adiponectin in PE. CONCLUSIONS Our findings showed higher levels of adiponectin and lower nitrite levels in PE compared to HP, and these levels were positively correlated in HP and in PE presenting lower concentrations of ADMA.

Correlations between circulating levels of adipokines and anti-angiogenic factors in women with BMI <30 and a late-onset preeclampsia

Preeclampsia (PE) is a pregnancy-specific disease, directly related to high rates of maternal–fetal morbidity and mortality worldwide. Upregulation of anti-angiogenic factors (soluble fms-like tyrosine kinase-1; sFLT-1 and soluble endoglin; sENG) have been suggested to trigger the maternal endothelial dysfunction observed in PE. Studies focusing on the role of adiponectin and leptin, in normal pregnancy as well as in complicated pregnancies, have revelated interesting findings due to the vascular actions of such adipokines. The aims of this study were to compare plasma concentrations of the adiponectin, leptin, sENG and sFLT-1 in preeclamptic (PE, n = 27) and healthy pregnant (HP, n = 36) and to evaluate possible correlations among these adipokines and anti-angiogenic factors. There were significant increases in all biomarkers in PE compared to HP (all p < 0.05). In PE group, there were positive strong correlations among adiponectin and leptin with sFLT-1 (r = 0.85 and r = 0.47, respectively) and sEng (r = 0.74 and r = 0.56, respectively). Moreover, we observed significantly correlation among body mass index (BMI) with adiponectin (r = −0.40) and with leptin (r = 0.51) in HP, but not in PE. Moreover, while a negative correlation between sFLT-1 and BMI (r = −0.60) was found in PE, no correlation was observed regarding sEng and BMI. In summary, our findings suggest the existence of a compensatory mechanism that occurs in an attempt to correct this angiogenic imbalance in order to restore the fetal development.

Role of adiponectin on antioxidant profile: evaluation during healthy and hypertensive disorders of pregnancy

Abstract The study of adipokines and oxidative stress has aided in understanding pre-eclampsia physiopathology. Therefore, our group aimed to evaluate the correlation between the adipokines (adiponectin and leptin) and the oxidative stress marker malondialdehyde–thiobarbituric acid reactive substances (MDA-TBARS) and antioxidant activity of plasma [ferric reducing ability of plasma (FRAP)] in healthy pregnant women and patients with gestational hypertension and pre-eclampsia. We found a significant negative correlation between MDA-TBARS and adiponectin (r = –0.40, p = 0.0042), suggesting a relationship between antioxidant levels and this adipokine in healthy pregnancies which is altered in patients with gestational hypertension or pre-eclampsia.

Características clínicas e laboratoriais de gestantes com pré-eclâmpsia versus hipertensão gestacional

PURPOSE To compare clinical and laboratory characteristics, obstetric and perinatal outcomes of patients with pre-eclampsia versus gestational hypertension. METHODS A retrospective study was carried out to analyze medical records of patients diagnosed with pre-eclampsia and gestational hypertension whose pregnancies were resolved within a period of 5 years, for a total of 419 cases. We collected clinical and laboratory data, obstetric and perinatal outcomes. Comparisons between groups were performed using the test suitable for the variable analyzed: unpaired t test, Mann-Whitney U test or χ2 test, with the level of significance set at p<0.05. RESULTS Were evaluated 199 patients in the gestational hypertension group (GH) and 220 patients in the pre-eclampsia group (PE). Mean body mass index was 34.6 kg/m2 in the GH group and 32.7 kg/m2 in the PE group, with a significant difference between groups. The PE group showed higher systolic and diastolic blood pressure and higher rates of abnormal values in the laboratory tests, although the mean values were within the normal range. Cesarean section was performed in 59.1% of cases of PE and in 47.5% of the GH group; and perinatal outcomes in terms of gestational age and birth weight were significantly lower in the PE group. CONCLUSION Women with gestational hypertension exhibit epidemiological characteristics of patients at risk for chronic diseases. Patients with pre-eclampsia present clinical and laboratory parameters of greater severity, higher rates of cesarean delivery and worse maternal and perinatal outcomes.

PP002. Study of polymorphisms of the adiponectin gene in women with gestational hypertension and preeclampsia

INTRODUCTION Adiponectin is involved in energy homeostasis by regulating glucose and lipid metabolism. Additionally, it presents anti-inflammatory and anti-atherosclerotic functions. Polymorphisms in adiponectin gene (ADIPOQ) can modulate the concentrations of adiponectin. The influence of these polymorphisms on the development of gestational hypertension (GH) and preeclampsia (PE) is unknown. OBJECTIVES The aim of this work was to examine the influence of polymorphisms in the gene ADIPOQ on the development of gestational hypertension and preeclampsia. METHODS PATIENTS AND METHODS We studied 401 pregnant women: 161 healthy pregnant (HP), 113 pregnant with gestational hypertension (GH) and 127 pregnant with preeclampsia (PE). Polymorphisms ADIPOQ -11391G>A (rs17300539), -11377C>G (rs266729), 45T>G (rs2241766) and 276G>T (rs1501299) were genotyped by allelic discrimination by PCR in real time. Haplotypes were inferred using the PHASE 2.1 program. RESULTS There were no statistically significant differences in allele and genotype frequencies of the polymorphisms studied. In the analysis of haplotypes, we observed small differences in haplotype frequencies between groups, however, none of these differences was statistically significant (P>0.05). CONCLUSION We found no association between the genotypic and allelic variants of the ADIPOQ gene polymorphisms with the development of gestational hypertension and preeclampsia.