Association of Lipids’ Metabolism with Vitamin D Receptor (rs10735810, rs222857) and Angiotensinogen (rs699) Genes Polymorphism in Essential Hypertensive Patients

BACKGROUND: Cardiovascular (CV) diseases are the most spread cause of mortality in the world. Essential arterial hypertension (EAH), as a major risk factor for the development of CV diseases, is a multifactorial disease involving environmental and genetic factors together with risk-conferring behaviors. AIM: The purpose of this study was to analyze lipid metabolism changes in patients with EAH depending on the Vitamin D receptor (VDR rs2228570 (aka rs10735810)) and angiotensinogen (AGT rs699) genes polymorphism. MATERIALS AND METHODS: The single-stage study involved 100 patients suffering from Stage 2 EAH, 1–3 degrees of blood pressure increase, high and very high CV risks, 21% (21) men, and 79% (79) women. The average age of patients was 59.86 ± 6.22 years old. The control group included 60 practically healthy individuals of an appropriate age and sex distribution. To examine the VDR gene (rs10735810, rs2228570) and AGT gene (rs699) polymorphism, a qualitative real-time polymerase chain reaction was made. The lipid metabolism was studied by determining the blood plasma content of total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), and triglycerides (TGs). RESULTS: Т allele of AGT gene is associated with reduced HDL-C level in men and increased TGs level in women. The EAH risk increases 4.5 times as much among the ТС-genotype carriers and lowered HDL-C level (odds ratio [OR] = 6.43; p = 0.01). The EAH risk increases as far as the HDL-C level reduction, irrespective of the VDR gene alleles condition 1.83 times (OR = 2.37; OR 95% confidence interval [CI]: 1.02–5.51; p = 0.04) and 1.9 times (OR=2.43; OR 95% CI: 0.99–5.97; p = 0.04). HDL-C reduction and LDL-C elevation in women increase the EAH risk 2.4 times (OR = 3.27; p = 0.01) and 1.24 times (OR = 3.67; p = 0.01), respectively. CONCLUSIONS: The EAH risk increases with a reduced HDL-C level in the TC genotype carriers of the AGT gene and irrespective of VDR gene polymorphic variants.

Associations of Gene Polymorphisms and Prognosis in Highly Adherent to Treatment Patients After Myocardial Infarction

Aim. To evaluate the influence of genetic factors on the risk of developing a combined endpoint, during a one-year supervision of patients, who had myocardial infarction and highly adherent to drug therapy.Material and methods. The research included 250 patients with high adherence to treatment with myocardial infarction, using the method of polymerase chain reaction we determined the polymorphisms Thr174Met and Met235Thr in the AGT gene, Arg389Gly and Ser49Gly in the ADRB1 gene, Ser447Ter in the LPL gene and Leu28Pro in the APOE gene, Trp212Ter and G681A in the CYP2C19 gene, and then we evaluated their influence on the prognosis.Results. A significant influence on the risk of developing combined endpoint was noticed for the polymorphism of CYP2C19 (G681A) gene. For the GA genotype of the CYP2C19 gene (G6881A), the OR of developing an unsuccessful outcome was 1.97 (95 % CI 1.05 — 3.69) (P = 0.03). For сarrier-state of A allele the OR was 1.46 (95 % CI 1.06 — 3.64) (P = 0.03). Conclusion. The results received indicate the need for individual approach for the choice of drugs from the group of inhibitors P2Y12-receptors for dual antiplatelet therapy, and if clopidogrel is chosen it is necessary to resolve the issue of pharmacogenetic testing for CYP2C19.

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.

Genomic Association Between Angiotensin-Converting Enzyme, Angiotensinogen And ARNT-Like Protein-1Genes With Susceptibility To Gestational Diabetes In Egyptian Women

Background and aims: Gestational diabetes mellitus (GDM) is well-defined as glucose intolerance first documented during pregnancy. In the present study, we examined the possible associations between I/D polymorphism of the angiotensin-converting enzyme (ACE) gene, the M235T variant of angiotensinogen (AGT) gene, and the rs7950226 polymorphism of the ARNT-like protein-1 (BMAL1) gene and the risk for GDM, in Egyptian pregnant women. Subjects and methods: This study recruited 160 GDM cases and165 controls. Genomic DNA was derived from peripheral blood leukocytes and ACE gene (I/D) genotyping was performed using the method of polymerase chain reaction (PCR) and the polymerase chain reaction-based restriction fragment length polymorphism (PCR-RFLP) was used for identifying the M235T variant of AGT gene and the rs7950226 polymorphism of the BMAL1. Results: The II, ID, and DD genotypes of the ACE gene have significant differences in GDM compared to controls (p= 0.000 and X2= 81.77). The M235T polymorphism of the AGT gene was increased with GDM risk. Furthermore, the AA genotype of the BMAL1 rs7950226 gene was significantly related to the GDM risk (P = 0.000 and X2= 52.82). The allele frequencies of the three variants vary significantly between women with gestational diabetes mellitus and control additionally; the risk factors of body mass index (BMI), family history, hypertension, and diabetes were significantly correlated with the GDM risk with a p-value of 0.001. Conclusion: The present study suggested significant associations between ACE (DD), AGT (TT), and BMAL1 rs7950226 (AA) gene polymorphisms with GDM susceptibility in Egyptian pregnant women and there was a possibility to identify that II + MM + GG as protective haplotypes and DD + TT + AA as risk haplotypes for GDM.

Associations Between M235T Polymorphism in the AGT Gene and Cancer: An Updated Systematic Review and A Meta-analysis

Background: We assessed the relationship between AGT gene M235T polymorphism and the susceptibility to cancer by performing an updated meta-analysis.Methods: This study retrospectively searched related articles in the electronic databases. Afterwards, we determined combined odds ratios (ORs) and related 95% confidence intervals (CIs) by the fixed- or random-effects model.Results: The present meta-analysis enrolled altogether 9 articles. On the whole, the relationship between AGT M235T polymorphism and the cancer risk was not significant among the entire population(TT vs MM:OR=1.28,95%CI=0.80-2.04;TM vs MM: OR=0.90, 95%CI = 0.53-1.52;Recessive model: OR= 1.13, 95%CI = 0.83-1.52; Dominant model: OR=0.93, 95%CI =0.55-1.57). But the relationship of AGT M235T polymorphism with the digestive cancer risk was significant upon subgroup analysis stratified according to cancer type (TT vs MM:OR=1.68,95%CI=1.11-2.54;TM vs MM: OR=1.34, 95%CI = 0.97-1.85;Recessive model: OR= 1.27, 95%CI = 0.95-1.70; Dominant model: OR=1.45, 95%CI =1.07-1.96).Conclusion: According to findings in the present meta-analysis, AGT M235T polymorphism may be possibly related to digestive cancer susceptibility.

Ethno-Territorial Distribution of the C174M and C235T Polymorphisms of the AGT Gene and C677T of MTGFR Gene in the Population of the Azerbaijan Republic

The prevention of hereditary diseases associated with gene and chromosomal disorders, in particular multifactorial-polygenic diseases is one of actual areas of medical genotyping. For the first time in the population of the Republic of Azerbaijan we have identified mutations C174T and C235T of the angiotensinogen gene and mutation C677T of the methylenetetrahydrofolate reductase gene both in the control group and among patients with diseases of the cardiovascular system. Reliable connections for the frequency of occurrence of polymorphism of the C174T and C235T alleles of the angiotensinogen gene and polymorphism of the C677T allele of the methylenetetrahydrofolate reductase gene were found with a statistical method. To identify the ethno-geographic relationship of the mutations C174T and C235T of the AGT gene for the population of the Azerbaijan Republic, we examined practically healthy individuals and patients with CVD. The composition of this group was multinational and corresponded to the main national and ethnic composition of the Republic. The distribution of the identified mutations C174T and C235T of the AGT gene, as well as the C677T polymorphism of the MTHFR gene among ethnic groups of the Azerbaijan Republic is identified as uneven.

Assessment of NF-κB Inhibitor (SN50) Effect on Adipose Tumor Necrosis Factor-Alpha and Angiotensinogen Secretion and Expression

Central adiposity is one of the significant determinants of obesity-related hypertension risk, which may arise due to the abdominal fat depot's pathogenic inflammatory nature. Pro-inflammatory cytokines and adipokines up-regulation through nuclear factor-kappa B (NF-κB) activation in adipose tissue has been considered an essential function in the pathogenesis of obesity-related hypertension. This study aimed to ascertain the NF-κB inhibitor (SN50) effect on TNF-α and angiotensinogen (AGT) secretion and expression in mediating the anti-inflammatory effect through its impact on NF-κB activity in humans adipose tissue. Primary human adipocytes were isolated from 20 subjects among 10 overweight and 10 obese with and without hypertension and treated with 10ng/ml LPS in the presence and absence of NF-κB inhibitor, SN50 (50μg/ml). TNF-α secretion and NF-κB p65 activity were detected in supernatants extracted from cultured cells treated and untreated with LPS (10ng/ml) and SN50 (50μg/ml) using enzyme-linked immunosorbent assay (ELISA). The western blot technique detected the protein of NF-κB p65 and AGT. Gene expression of TNF-α and AGT was detected in cells and performed using quantitative real-time polymerase chain reaction (RT-PCR). Treatment of AbdSc adipocytes with LPS (10ng/ml) caused a significant increase in NF-κB p65 among overweight and obese subjects with and without hypertension (P= 0.001) at 24 hours incubation. In contrast, SN50-NF-κB inhibitor causes a reduction of NF-κB p65 in overweight (P= <0.001) and obese subjects with and without hypertension (P= 0.001) at 24 hours incubation. Treatment of AbdSc adipocytes with 10ng/ml LPS caused a significant increase in TNF-α secretion in overweight and obese subjects at all-time points (P= <0.001), whereas SN50 leads to a decrease in TNF-α secretion at 3 and 12 hours incubation. Treatment of AbdSc adipocytes with LPS (10ng/ml) caused increased TNF-α and AGT gene expression twofold compared with untreated cells, whereas, in the presence of SN50, it reduces mRNA AGT levels in both groups. Taken together, these adipokines with NF-κB activation may represent essential biomarkers to evaluate hypertension risk and to provide insight into the pathogenesis of obesity-related hypertension.

CRISPR/Cas9 Mediated Deletion of the Angiotensinogen Gene Reduces Hypertension

Hypertension is a major contributor to the global burden of disease. Unfortunately, hypertension is controlled in less than one-fifth of patients worldwide due to either failure to treat or lack of compliance to medication. An ideal therapy would be administered one time only and yield lifelong blood pressure control. We investigated our hypothesis that CRISPR/Cas9 (clustered regularly interspaced short palindromic repeats/clustered regularly interspaced short palindromic repeat–associated 9)-mediated disruption of a key gene in the renin-angiotensin system, AGT (angiotensinogen), specifically in the liver, would result in sustained and possibly lifelong reduction in blood pressure. We demonstrated in vitro that the CRISPR/Cas9 system led to a significant reduction in AGT expression in hepatocytes. Delivery of the CRISPR/Cas9 system into the liver via the hepatocyte-targeting adeno-associated virus 8 reduced both AGT expression (40% decrease) and circulating AGT levels (30% decrease). In the SHR (spontaneously hypertensive rat) model of hypertension, CRISPR/Cas9-mediated loss of AGT expression reduced blood pressure in adult animals with established hypertension and prevented the spontaneous development of hypertension in young SHR. Moreover, reductions in blood pressure were prolonged and sustained up to 1 year of follow-up. In addition, the partial disruption of the hepatic AGT gene was sufficient to control hypertension but did not affect the homeostatic response to cardiovascular stress such as sodium depletion and furosemide. In summary, we have demonstrated that targeting the CRISPR/Cas9 system to hepatic AGT results in sustained reduction of blood pressure and is a potential therapy to achieve sustained and possibly lifelong control of human hypertension.

AGT M235T Polymorphism and Arterial Hypertension in Patients with Chronic Obstructive Pulmonary Disease

Background: Chronic obstructive pulmonary disease (COPD) represents an important public health challenge. It is a major cause of chronic morbidity and mortality throughout the world. Arterial hypertension (AH) is considered one of the principal COPD-associated comorbidities. Objective: The study was aimed to assess the role of angiotensinogen (AGT) gene polymorphism in occurrence of AH in patients with COPD. Methods: The study was conducted among 96 patients of them, Group 1 (25 individuals with COPD), Group 2 (23 individuals with AH), Group 3 (28 individuals with COPD and AH). The control group consisted of the 20 healthy subjects. I/D genotypes of AGT were determined by polymerase chain reaction amplification. Plasma AGT activity was determined photometrically by a commercially available kit. Results: The results of the study have not demonstrated any significant impact of alleles of AGT genes on occurrence of such disease as COPD, AH and combinations thereof. However, analysis of odds ratio has demonstrated the presence of a trend towards a protective role of the M allele of the AGT gene concerning occurrence of COPD, AH and their combinations (OR=0.90, OR=0.71 and OR=0.56, respectively). At the same time, the presence of the T allele of the AGT gene may increase the risk for occurrence of the above mentioned disease (OR=1.11, OR=1.4 and OR=1.79, respectively). Conclusion: The study suggests that the presence of Т allele of the AGT gene at position 235 of the peptide chain both in homozygous and heterozygous state may increase the risk for AH in patients with COPD. Bangladesh Med Res Counc Bull 2020; 46(3): 176-183