VEGF and eNOS genes polymorphism features in patients with diabetes mellitus with and without initial non-proliferative diabetic retinopathy

The endothelial NO synthase (eNOS) and vascular endothelial growth factor (VEGF) imbalance and the polymorphism of these genes may be the predisposition for diabetic retinopathy (DR) development and progression.The aim: to analyze VEGF (rs699947 and rs3025039) and eNOS (rs2070744) genes polymorphism and their combinations in patients with type 2 diabetes mellitus (DM2) with and without initial non-proliferative DR.Materials and methods. The study included 200 patients with type 2 diabetes (155 women and 45 men, age – 43–70 years): 111 people without and 89 people with DR. The polymorphism of the regulatory regions of VEGF (rs699947 and rs3025039) and eNOS (rs2070744) genes was studied using restriction fragment length polymorphism analysis and TaqMan Real-Time PCR by. Statistical processing was carried out using the software packages Statistica 10.0, SPSS Statistics 23 and the package of original programs for volumetric processing of bioinformation.Results. The VEGF-2578 heterozygosity and two complex genotypes – VEGF-2578CA:VEGF+936CC and NOS3-786CT:VEGF-2578CA:VEGF+936CC – signifi cantly decreased in patients with DR. The predisposition to early DR development to minor genotype of eNOS gene in the NOS3-786CC:VEGF+936CT complex and signifi cantly decreased the homozygous wild-type eNOS genotype in DM2 patients with ophthalmopathology were shown. NOS3-86TT:VEGF2578AA genotype signifi cantly decreased in group with retinopathy developing and the glycated hemoglobin high level.Conclusion. Along with the clinical risk factors for the development of DR in DM2, the genetic polymorphism of the regulatory regions of the genes analyzed by us has a signifi cant weight. When analyzing potential genetic markers, it is important to consider possible joint epistatic/hypostatic effects. The complex analysis of polymorphic gene can help early prognosis of the DR development.

The association of T786C and G894T polymorphisms of eNOS gene with diabetic retinopathy in Greece

Purpose To investigate whether eNOS T786C (rs2070744) and G894T (rs1799983) gene polymorphisms are associated with diabetic retinopathy in Greek diabetic patients. Materials 271 patients with type-2 diabetes mellitus participated in our study; 130 suffered from diabetic retinopathy and 141 not. All the patients underwent a complete ophthalmological examination, while clinical and demographic data were assessed. Furthermore, they were genotyped for rs2070744 and rs1799983 single nucleotide polymorphisms of eNOS gene. Results Regarding the clinical and demographic data, no significant differences were detected between the studied groups, except for hemoglobin A1c levels and the frequency of insulin treatment (higher in patients with diabetic retinopathy). The frequency of rs1799983 GT genotype was significantly elevated in patients with diabetic retinopathy (55% vs. 40%, P = 0.011) and was associated with a 2-fold increased risk of developing retinopathy (OR 1.92, 95% CI 1.16–3.17). Furthermore, we demonstrated that the aforementioned genotype was significantly and independently associated with increased odds for retinopathy onset in diabetic subjects (OR 2.23, 95% CI 1.28–3.90, P = 0.005), regardless of the impact of other confounders. Conclusions We documented that rs1799983 GT genotype could be recognized as an independent risk factor of retinopathy in Greek patients with type-2 diabetes mellitus, while no role for rs2070744 polymorphism was identified. Further research in different ethnic groups will clarify the exact association of these polymorphisms with the risk for diabetic retinopathy development.

Endothelial nitric oxide synthase G894T, intron 4 VNTR, and T786C polymorphisms in retinopathy of prematurity

BACKGROUND: Our objective in this study was to assess the association between eNOS gene, that achieves synthesis of nitric oxide especially in the endothelial cells known to have an important role in angiogenesis and vasculogenesis, G894T, intron 4 VNTR (27-bp repeat) and T786C functional polymorphisms and retinopathy of prematurity (ROP), which is an important cause of morbidity in premature or low birth weight babies. METHODS: A total of 139 babies who were followed up in our neonatal intensive care unit because of premature birth in our hospital or admitted to our unit. 69 of them had retinopathy of prematurity and comprised the patients group. The remaining 70 babies who did not have ROP comprised the control group. An additional of 1 ml of blood samples were drawn from babies who were in the study groups during routine laboratory analysis. eNOS gene polymorphisms were determined by using polymerase chain reaction method. RESULTS: eNOS G894T, intron 4 VNTR and T786C gene polymorphisms did not differ between the patient and control groups (p > 0.05). Using logistic regression analysis; while gender did not differ between two groups; gestational age, birth weight, time on mechanical ventilation differ between two groups. After adjustment for variables other than eNOS gene polymorphisms, we found no significant difference in the genotype distribution of eNOS G894T, intron 4 VNTR and T786C polymorphisms (p > 0.05). CONCLUSION: We observed no association between ROP and eNOS gene polymorphisms but needs more investigation.

Assessment of calcium antagonist therapy protective effect on the thickness of the intima-media complex in patients with arterial hypertension

Aim. To evaluate the connection of calcium antagonist (amlodipine) therapy with the dynamics of the intima-media complex thickness in patients with arterial hypertension (AH), depending on genetic polymorphism.Methods. The study included representatives of the indigenous nationality (the Shors) – 901 people, of which a group of 367 people with hypertension was identified. The prospective stage of observation included 234 people who did not receive antihypertensive therapy. Based on the prescription of calcium antagonists, patients with hypertension were divided into two groups. Gene polymorphism was tested by polymerase chain reaction.Results. In the Shor cohort, the regression of the intima-media complex thickness of the carotid arteries was observed more often in hypertensive patients who received calcium antagonists if to compare them with those who did not take the drug [OR = 2.30]. In addition, the decrease in the atherosclerotic process is associated with the genotype carriage: I/I of the ACE gene [OR = 9.42], T/C of the AGT gene [OR = 3.52], 4b/4b and 4b/4a of the eNOS gene [OR = 2.26 and OR = 3.75], C/C of the MTHFR gene [OR = 2.62].Conclusion. Pharmacogenetic aspects are valuable from the point of view of an individual approach and obtaining the most pronounced pharmacological response in order to slow down the processes of vascular wall remodeling in patients with hypertension.

Clinical and Prognostic Value of Calcium and Phosphorus Levels as Possible Markers of Endothelial Dysfunction in Preterm Infants

The system maintaining a sufficient level of calcium and phosphorus is formed only by the end of the first month of life, and in pretermure infants even later. These elements play a crucial role in many physiological processes. They also affect vascular function, endothelium, and endothelial nitric oxide (NO) synthase. Therefore, the aim of our study was to analyze the relationship of serum phosphorus and calcium levels with markers of endothelial dysfunction and features of the early neonatal period. The study included 37 preterm infants that received standard clinical, laboratory, and instrumental examination. As markers of endothelial dysfunction, the 4a/4b polymorphism of the eNOS gene was investigated and quantitative measurement of NO metabolites in urine was performed. Phosphorus and calcium levels in 37 preterm infants on the first day were 4.06 ± 1.88 mg/dL and 1.89 ± 2.08 mg/dL, and on the six day 4.89 ± 2.02 mg/dL and 1.86 ± 1.80 mg/dL, respectively. Combined hypocalcemia and hypophosphatemia were found in 27 (73.0%) of 37 examined infants. The presence of severe condition in early neonatal period was positively correlated with calcium levels on the first and sixth days after birth. There were determined positive correlation between calcium and phosphorus levels on the first day after birth and severity of clinical symptoms in neonatal period and birth gestational age. Serum calcium level in preterm birth infants was positively correlated with respiratory failure on the sixth day. Correlation between calcium and phosphorus serum levels was observed for severe and stable infants on the first day, but on the six day only in preterm birth infants with stable condition. We have not found an association between phosphorus and calcium levels and markers of endothelial dysfunction in preterm birth infants. Serum phosphorus and calcium levels and studied markers of endothelial dysfunction have been found to be independent markers of the risk of developing a complicated early neonatal period in preterm infants.

Study of genes involved in angiogenesis and metabolic processes of peripheral blood lymphocytes in patients with chronic endometritis

Objective. To study the polymorphisms of the genes involved in angiogenesis and in metabolic processes, to assess the level of lymphocytes in patients with chronic endometritis and practically healthy women of reproductive period. Materials and methods. 86 patients were examined; DNA regions of the genes eNOS 1799983 (Glu298Asp), PPARA (G2528C), ApoE rs429358 (Cys130Arg), MTHFR (C677T, A1298C) were used as primers; blood lymphocytes (CD3+, CD4+, CD8+, CD19+, CD95+) were assessed. Results. Statistically significant differences in gynecological and chronic somatic pathology were obtained in patients with chronic endometritis; they more often than practically healthy women had polymorphisms of the genes ApoE rs429358, eNOS1799983, PPARA (G2528C); patients with chronic endometritis more often had dysregulation of the immune system in the form of insufficiency of the cellular effector link of immunity and changes in the PPARA, ApoE, eNOS gene. Attention was drawn to the obtained relationships of polymorphic genes and clinical manifestations in patients with chronic endometritis, in particular, with a history of non-developing pregnancy in anamnesis, there was more often detected polymorphism of the ApoE gene, with abnormal uterine bleeding polymorphism of PPARA, with chronic inflammatory pathology of the gallbladder polymorphism of the MTHFR gene. Conclusions. The prevalence of polymorphism of the genes eNOS 1799983 (Glu298Asp), PPARA (G2528C), ApoE rs429358 (Cys130Arg), MTHFR (C677T, A1298C) was obtained in patients with chronic endometrial inflammation compared with practically healthy participants in the study. Insufficiency of the cellular effector link of immunity was revealed in the majority of patients with ChE and an association with allele C genotypes G/C and C/C of PPARA 4253778 gene, with allele C genotypes G/C and C/C of ApoE42935 gene, with allele C genotypes G/C and C/C of eNOS 1799983 gene and G/C genotype of MTHFR gene (C677T, A1298C).

Specificity of blood pressure numbers in Holter monitoring depending on gene polymorphism among residents of Ternopil region afflicted with essential arterial hypertension

The aim of this work is to determine specificities of blood pressure (BP) numbers during the day depending on polymorphism of the A1166C-gene of angiotensin II receptor type I and T786C-promoter of the endothelial NO-synthase gene among residents of Ternopil region afflicted with essential arterial hypertension.We have examined 86 patients with arterial hypertension who were treated and examined in the therapeutic department of the Central District Hospital in Kozova, aged from 45 to 76 years. All patients were measured for body weight and height, office blood pressure; also they were checked by Holter monitoring and electrocardiography (ECG), as well as examined for polymorphism of endothelial NO synthase genes and angiotensin II type 1 receptor genes. Statistical processing of the obtained data was performed with the help of the analytics software package including Statistica 8.0 (StatSoft Inc., USA) and Microsoft Office Excel-2003.The research revealed that patients with CC genotype A1166C-gene of angiotensin II receptor of the first type had a significantly higher level of systolic blood pressure (SBP) and diastolic blood pressure (DBP) – average numbers per 24 hours, day and night – compared to patients with genotype AA(p<0,05). Hereby, no significant distinctions in blood pressure variability were found.Patients with CC of the T786C promoter of the eNOs gene had higher SBP and DBP values at all times of the day compared to individuals with the TT genotype (p <0.05). Increased levels of variability of DBP during the day, as well as SBP and DBP at night, were observed among patients with CC genotype, as compared with the control group (p <0.05).Among patients with C-allele of both studied genes, there’s a high frequency of circadian rhythm disorders with predominance of the “non-dippers” pathological type.

Germline GATA2 variant disrupting endothelial eNOS cell function and angiogenesis can be restored by c-Jun/AP-1 upregulation

GATA2 is a transcription factor with key roles in hematopoiesis. Germline GATA2 gene variants have been associated with several inherited and acquired hematologic disorders, including myelodysplastic syndromes. Among the spectrum of GATA2 deficiency-associated manifestations thrombosis has been reported in 25% of patients, but the mechanisms are unknown. GATA2 was shown to be involved in endothelial nitric oxide synthase (eNOS) regulation and vascular development. We assessed eNOS expression and angiogenesis in patients with GATA2-deficiency. Platelets and blood outgrowth endothelial cells (BOEC) from GATA2-variant carriers showed impaired NO-production and reduction of eNOS mRNA and protein expression and of eNOS activity. GATA2 binding to the eNOS gene was impaired in BOEC from GATA2-deficiency patients, differently from control BOEC. GATA2-deficiency BOEC showed also defective angiogenesis, which was completely restored by treatment with the NO-donor S-nitroso-N-acetylpenicillamine (SNAP). Atorvastatin, but not resveratrol, largely restored eNOS expression, NO biosynthesis and neoangiogenesis in GATA2-deficient BOEC by a mechanism involving increased expression of the eNOS transcription factor AP-1/c-JUN, replacing GATA2 when the latter is inactive. Our results unravel a possible thrombogenic mechanism of GATA2 mutations, definitely establish the regulation of eNOS by GATA2 in endothelial cells and show that endothelial angiogenesis is strictly dependent on the eNOS/NO axis. Given the ability of atorvastatin to restore NO production and angiogenesis by GATA2-deficient endothelial cells, the preventive effect of atorvastatin on thrombotic events and possibly on other clinical manifestations of the syndrome related to deranged angiogenesis should be explored in patients with GATA2-deficiency in an ad hoc designed clinical trial.