AT1 Receptor Gene Research Articles

Correlation between AT1R gene polymorphism and epilepsy secondary to cerebral infarction.

To explore the correlation between angiotensin II type 1 receptor (AT1R) gene polymorphism and epilepsy secondary to cerebral infarction and its significance for the diagnosis of this disease. A total of 200 patients with epilepsy secondary to cerebral infarction were enrolled from our hospital as observation group, and 200 patients without epilepsy after cerebral infarction as control group. Genomic deoxyribonucleic acids (DNAs) were extracted from the peripheral blood of the subjects, and the polymorphic regions at AT1R gene loci rs380400, rs1799870, rs12721273, and rs55707609 were amplified via polymerase chain reaction (PCR) and sent to the company for sequencing. The concentration of farnesyl diphosphate synthase (FDPS) was determined using enzyme-linked immunosorbent assay (ELISA) kit, and activated partial thromboplastin time (APTT) and prothrombin time (PT) were measured in the Laboratory Department. There were no differences in the allele distributions at AT1R gene loci rs380400 (p=0.070), rs179987 (p=0.0.280), and rs55707609 (p=0.046), but in the allele distribution at rs12721273 (p=0.001) between control group and observation group, and observation group exhibited a significantly lower frequency of allele G in cerebral infarction patients than control group [153 (0.383) vs. 198 (0.495)]. The frequency of genotype GT at rs12721273 was lower [71 (0.355)] and that of genotype TT was evidently higher [88 (0.440)] in observation group (p=0.000). Control group showed a notably lower frequency of genotype AA [47 (0.235)] and a markedly higher frequency of genotype AT [110 (0.550)] at rs55707609 (p=0.000). Observation group exhibited a substantially lower frequency of recessive model AG+GG [128 (0.640)] (p=0.037), and a notably higher frequency of homozygous model AA [72 (0.360)] (p=0.048) at AT1R gene locus rs380400, a remarkably lower frequency of dominant model GG+GT [112 (0.560)] (p=0.002) at rs12721273, and a significantly lower frequency of recessive model AT+TT [126 (0.630)] (p=0.000) and a considerably lower frequency of heterozygous model AT [84 (0.420)] (p=0.026) at rs55707609. The frequencies of AT1R gene haplotypes ACGA (p=0.001), ACGT (p=0.045), ACTT (p=0.000), ATTT (p=0.048), GCTA (p=0.000), and GTGA (p=0.005) in observation group were distinctly higher than those in control group, and the frequencies of the haplotypes ACTA (p=0.000) and ATTA (p=0.029) were evidently lower than those in control group. The loci rs12721273 and rs1799870 showed a significant association (D'=0.783), and APTT was considerably correlated with genotype AG at rs380400 (p=0.042), PT with genotype CC at rs1799870 (p=0.002) and FDPS with genotype AA at rs55707609 (p=0.015). The polymorphisms of AT1R gene loci rs380400, rs1799870, rs12721273, and rs55707609 are correlated with the susceptibility to epilepsy secondary to cerebral infarction.

Correlation between gene polymorphism in angiotensin II type 1 receptor and type 2 diabetes mellitus complicated by hypertension in a population of Inner Mongolia

BackgroundThe role of angiotensin II type 1 receptor (AT1R) as a key player in type 2 diabetes mellitus (T2DM) complicated with hypertension remains controversial. The present case-control study systematically investigated the association between gene the correct variation type in the angiotensin II type 1 receptor (AT1R) gene and type 2 diabetes mellitus complicated with hypertension in the Han population from the Inner Mongolia region, China.MethodHere, state which variants were analysis, including age, occupation, triglyceride, systolic, diastolic, sex, culture, marital status, smoking, alcohol, BMI (body mass index), SBP (systolic blood pressure), DBP (diastolic blood pressure), TG (triglyceride), TC (total cholesterol), HDL-C (high-density lipoprotein cholesterol), LDL-C (low-density lipoprotein cholesterol), FPG (fasting plasma glucose). Genomic DNA was extracted from samples from 202 type 2 diabetic patients with hypertension and 216 type 2 diabetic patients without hypertension.ResultsNon-conditional regression analysis showed that in comparison with the TT genotype, the presence of the CC genotype for the T573 site of the AT1R gene increased the risk for diabetes mellitus complicated with hypertension by 3.219-fold (OR = 3.219, 95% CI: 1.042–9.941, P = 0.042). The results from multivariate linear regression analysis suggested the rs5182 polymorphism in the AT1R gene to be associated with diastolic blood pressure (P = 0.032). No other associations were found between the incidence of disease and the correct variation type at other sites of the AT1R gene.ConclusionsOur results suggest that the rs5182 polymorphism in the AT1R gene is associated with diabetes complicated by hypertension in the Han population of Inner Mongolia.

Associations Between Polymorphisms of Endothelial Nitric Oxide Synthase, Matrix Metalloproteinase 3, Angiotensinogen, and Angiotensin II Type 1 Receptor and Risk of Restenosis After Percutaneous Coronary Intervention: A Meta-analysis

PurposePrevious studies have reported controversial results regarding the risk of restenosis with polymorphisms of endothelial nitric oxide synthase (eNOS), matrix metalloproteinase 3 (MMP-3), angiotensinogen (AGT), and angiotensin II type 1 receptor (AT1R) after percutaneous coronary intervention (PCI). This study aimed to summarize the association between these polymorphisms and risk of restenosis after PCI. MethodsWe searched the electronic databases of PubMed, Embase, Cochrane's Library, and ClinicalTrials.gov for studies on the association of eNOS, MMP-3, AGT, and AT1R polymorphisms with restenosis. FindingsA total of 17 studies (7781 patients) were analyzed, including 5 studies on eNOS G298A (n = 912), 5 studies on MMP3 5A/6A (n = 4519), 6 studies on AGT M235T (n = 1801), and 7 studies on AT1R A1166C (n = 2477). For the G298A variant of the eNOS gene, the allele odds ratio (OR) was 1.685 (95% CI, 1.269–2.338; P < 0.001), the heterozygote OR was 2.144 (95% CI, 1.490–3.085; P < 0.001), the dominant OR was 2.078 (95% CI, 1.462–2.954; P < 0.001), and the overdominant OR was 0.496 (95% CI, 0.348–0.706; P < 0.001). For the 5A/6A variant of the MMP3 gene, the heterozygote OR was 0.839 (95% CI, 0.722–0.975; P = 0.022), the dominant OR was 0.846 (95% CI, 0.733–0.976; P = 0.022), and the overdominant OR was 1.141 (95% CI, 1.001–1.301; P = 0.049). For the M235T variant of the AGT gene, the heterozygote OR was 1.594 (95% CI, 1.179–2.155; P = 0.002), the dominant OR was 1.437 (95% CI, 1.077–1.918; P = 0.014), and the overdominant OR was 0.694 (95% CI, 0.555–0.869; P = 0.001). Positive results were observed in the AT1R gene A1166C polymorphism under 3 models (homozygote OR = 2.009; 95% CI, 1.433–2.816; P < 0.001; recessive OR 1.874; 95% CI, 1.353–2.595; P < 0.001; and dominant OR = 1.350; 95% CI, 1.105–1.649; P = 0.003). ImplicationsThe G298A variant of eNOS, the 5A/6A variant of MMP3, the M235T variant of AGT, and the A1166C variant of A1TR may increase the risk of restenosis after PCI.

Antagonism of Angiotensin II AT1 Receptor and Silencing of CD44 Gene Expression Inhibit Cardiac Fibroblast Activation via Modulating TGF-&amp;lt;i&amp;gt;β&amp;lt;/i&amp;gt;1/Smad Signaling Pathway

Angiotensin II (Ang II) is known to elicit cardiac fibrosis by activating the AT1 receptor and CD44 expression in the in vivo model. However, the cellular/molecular mechanisms underlying cardiac fibrosis are still not well understood. This study examines the roles of the AT1 receptor and CD44 gene expression in collagen synthesis through Ang II stimulated cardiac fibroblasts. Fibroblasts were isolated from the neonatal rat hearts; the activation of fibroblasts was evaluated using the assays of cell viability and migration, and silencing of CD44 gene expression was conducted with small interfering RNA (siRNA). Results showed that Ang II significantly increases the cell proliferation and migration in a dose-dependent manner. Upon activation, the protein levels of TGF-β1, Smad2, Smad4 and collagen I were significantly increased (all p by the AT1 receptor blocker, telmisartan (all p β1 as demonstrated by Pearson correlation analysis (r = 0.955, p < 0.01). Gene transfection of fibroblasts with Ad-CD44 siRNA, as evidenced by low levels of CD44 mRNA and protein, significantly reduced the production of collagen I. In summary, these results indicate that the proliferation, migration and collagen production from Ang II activated cardiac fibroblasts are potentially mediated by the AT1 receptor and CD44. Such a signaling mechanism could be crucial for the production of collagen and the development of tissue fibrosis in the heart.

Genetic Polymorphism of Angiotensin II Type 1 Receptors and Their Effect on the Clinical Outcome of Captopril Treatment in Arab Iraqi Patients with Acute Coronary Syndrome (Mid Euphrates).

Genetic variation in the angiotensin II type 1 receptor (AT1R) has an important effect on the outcome of acute coronary syndrome (ACS) initiated treatment with captopril. This study aims to investigate the impact of genetic polymorphism of AT1R (rs5186 and rs275651) on the ACS outcome in Iraqi patients treated with captopril. A total of 250 Iraqi individuals with ACS were included in this case-control study and they were divided into two study groups; Study group 1 included 125 participants who were prescribed captopril, 25mg twice daily and study group 2 included 125 participants who received no captopril as part of their ACS treatment (control study). The AT1R gene (rs5186) CC genotype was found to be associated with ST-elevation myocardial infarction (STEMI) (Odd's ratio (O.R) = 1.2, P = 0.7), while AC was associated with Non-ST-elevation myocardial infarction (NSTEMI) and unstable angina (UA) (O.R = 1.2, P = 0.8). AC genotype is more prone to have Percutaneous coronary intervention (PCI) after ACS attack (O.R = 1.2, P = 0.6). CC genotype had a risk to get less improvement (O.R = 1.6, P = 0.5), so might require higher doses of captopril during acute coronary insult. The AT1R gene (rs275651) AA genotype was associated with UA (O.R = 1.3, P = 0.9). AA and AT genotypes were more prone to have PCI after ACS attack (O.R = 3.9 P = 0.2, O.R = 3.5, P = 0.3 respectively) and thus requiring higher doses of captopril. We conclude that the AT1R rs5186, rs275651 genetic polymorphisms might partially affect the clinical outcome of ACS patients treated with captopril and might have captopril resistance which requires higher doses.

Risk and severity of psoriasis vulgaris in relation to angiotensin II type 1 receptor gene polymorphism and metabolic syndrome.

BackgroundPsoriasis vulgaris is a chronic inflammatory and proliferative skin disease, characterized by the formation of itchy, erythematous skin patches or plaques. Patients with psoriasis are at an increased risk of developing metabolic syndrome, including obesity, hypertension, diabetes, and atherosclerosis. Recently, angiotensin II (Ang II) has been reported to be associated with the development of psoriasis. Ang II not only increases the blood pressure but is also a potent proinflammatory modulator and functions through interaction with angiotensin II type 1 receptor (AT1R). Moreover, it is hypothesized that the AT1R gene expression could be correlated with the severity of psoriasis and/or metabolic syndrome.AimWe examined the association of Ang II type 1 receptor (AT1R) A1166C gene polymorphisms and metabolic syndrome with the severity of psoriasis.Patients and methodsThe present case-control study included 25 patients with psoriasis vulgaris and 25 healthy subjects in Egypt. The psoriasis lesions in the patient group were assessed using the psoriasis area and severity index (PASI) score. The AT1R polymorphism A1166C (rs5186) was studied using restriction fragment length polymorphism (RFLP) and polymerase chain reaction (PCR) amplification of the gene from the whole blood sample in both groups. Serum lipid profile and blood sugar levels were assessed post 12 h and 8 h fasting, respectively, in both groups. The severity of metabolic syndrome was evaluated using the severity score.ResultsThe results of the present study demonstrated that the AT1R A1166C gene polymorphisms increased the risk of developing psoriasis in the Egyptian population. We found that 70% of patients with AC genotype and 100% of patients CC genotype reported a PASI score >20 and were considered to be severe cases with a statistically significant difference as compared with patients with AA genotype (p=0.003). In addition, a high statistically significant difference (p=0.001) existed among AT1R genotypes with respect to the percentage of metabolic syndrome in psoriasis patients. Similarly, a statistically significant difference (p=0.004) among AT1R genotypes with respect to metabolic score was found, with the highest level of score and percentage observed in patients with CC genotype than in patients with AC genotype. The lowest level was present among those with AA genotype.ConclusionPatients with psoriasis expressing the C allele of AT1R1166 are susceptible to developing metabolic syndrome and have higher PASI scores as compared with patients carrying the A allele.

Blockade of the AT1 receptor and silencing of CD44 gene expression are associated with inhibition of angiotensin II‐activated cardiac fibroblasts

ObjectiveWe have previously demonstrated that the stimulation of fibroblasts by angiotensin II (Ang II) contributes to cardiac fibrosis via transforming growth factor beta (TGF‐β1)‐mediated collagen production. The cellular/molecular mechanism responsible for fibroblast activation, however, is not well understood. CD44, a cell‐surface glycoprotein, is associated with cell‐cell interactions, cell adhesion and migration. This study examines the roles of Ang II AT1 receptor and CD44 gene expression in the activation of cardiac fibroblasts.MethodsCardiac fibroblasts were isolated from the neonatal rat hearts, the assays of cell viability and migration were selected to demonstrate the activation of fibroblasts, and silencing of CD44 gene expression on fibroblasts was conducted with small interfering RNA (siRNA).ResultsImmunofluorescence staining with vimentin and discoid in domain receptor‐2 revealed the purity of isolated fibroblasts. Ang □ significantly reduced the cell viability and increased the cell migration in a dose‐dependent manner with a maximal stimulation as seen at 50 nM. In coincidence with fibroblast activation, mRNA and protein level of CD44 were up‐regulated and the protein levels of TGF‐β1, Smad2/4 and collagen I were significantly increased (all p&lt;0.05 vs. unstimulated cells). Addition of Ang II AT1 receptor blocker, telmisartan into the cells abolished Ang II‐elicited stimulating effects on these parameters measured (all p&lt;0.05 vs. Ang II activated cells). Consistent with data showing that there is a linear correlation between CD44 and TGF‐β1 as demonstrated by Pearson correlation analysis (r=0.955, p&lt;0.01), gene transfection of fibroblasts with Ad‐CD44 siRNA as identified by mRNA and protein levels of CD44 demonstrated a significant attenuation in the production of collagen I, suggesting a role of CD44 in the activation of fibroblasts.ConclusionsThese results indicate that cardiac fibroblasts, in response to Ang II stimulation, express high levels of the AT1 receptor and CD44, which reduce the cell viability and facilitate cell migration. Such a signaling mechanism could be crucial for leading to the deposition of collagen and the production of cardiac fibrosis.Support or Funding InformationThis study was supported in part by grants from the Mercer University School of Medicine, the Medcen Community Health Foundation, Georgia and the National Natural Science Foundation of China (81470436).This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Angiotensin II type 2 receptor gene polymorphisms and serum angiotensin-converting enzyme level in Egyptian children with systemic lupus erythematosus.

There are no reports about the association of angiotensin II type 2 receptor ( AT2R) gene polymorphisms and susceptibility to systemic lupus erythematosus (SLE) in children. The objective of this research is to study AT2R gene polymorphisms in exon 3 (C1593A) and intron 1 (A1675G) in Egyptian children with SLE and its correlation with disease manifestations and serum angiotensin-converting enzyme (ACE) level. Typing of AT2R gene polymorphisms was conducted in 123 children with SLE in comparison with 100 healthy controls using the restriction fragment length polymorphism method. Significant differences were found between SLE patients and controls for A-containing genotypes (CA + AA) and A-allele frequencies of AT2R in exon 3 (C1593A) ( p = 0.01, odds ratio (OR) = 2.5, 95% confidence interval (CI) = 1.3-5.05; p = 0.01, OR = 2.2, 95% CI = 1.2-4.1, respectively). G-containing genotypes (AG + GG) and G allele of AT2R in intron 1 (A1675G) were more frequent in SLE patients compared to controls ( p = 0.01, OR = 2.3, 95% CI = 1.2-4.5; p = 0.02, OR = 2.1, 95% CI = 1.2-3.7, respectively). Serum ACE level was significantly higher in SLE patients than in controls ( p < 0.001). There was no association between AT2R gene polymorphisms and ACE level in serum. Moreover, there was no association between AT2R gene polymorphisms and SLE clinical manifestations. AT2R gene polymorphisms can be considered risk factors for SLE development in Egyptian children.

Association of angiotensin II type 1 receptor gene A1166C polymorphism with steroid sensitivity in children with nephrotic syndrome

Introduction: In children, nephrotic syndrome is usually idiopathic. Cases are considered as minimal change disease until proven otherwise. The children who respond to steroid are called steroid sensitive, and the children who do not respond to steroid are called steroid resistant. Objectives: According to the role of genetic in effectiveness of steroid therapy of children with idiopathic nephrotic syndrome, we designed this study to investigate the role of angiotensin II type 1 receptor (AT1R) gene A1166C polymorphism in such conditions. Patients and Methods: This single center study was conducted on Iranian children in Hazarat Ali Asghar hospital, Tehran, Iran. A total of 40 patients (including 18 steroid resistant and 22 steroid sensitive patients) were selected. Total DNA samples were taken from peripheral blood. Polymerase chain reaction (PCR) was used for genotyping. Results: In this study, the association of AT1R gene A1166C polymorphism with groups including steroid sensitive and steroid resistant groups was not significant (P>0.05). We also found a significant difference of systolic blood pressure between steroid sensitive and steroid resistant group with more values in steroid resistant group (P=0.0327). Conclusion: This low power single center study could not show any significant association for this polymorphism in Iranian population. Resistance to steroid therapy was associated with higher systolic blood pressure.

MAPK/AP-1 pathway activation mediates AT1R upregulation and vascular endothelial cells dysfunction under PM2.5 exposure

Acute and chronic exposure to particulate matter (PM) 2.5 is associated with adverse health effect upon the cardiovascular (CV) system. However, the molecular mechanism by which PM2.5 evokes CV injuries has not been fully clarified. In our recent report, we demonstrate that exposure to PM2.5 leads to elevation of circulating angiotensin II (ANGII) levels and local expressions of angiotensinogen (AGT, the precursor of ANGII), angiotensin-converting enzyme (ACE) and ANGII type 1 receptor (AT1R) in the vascular endothelial cells, which subsequently instigates the oxidative stress and proinflammatory response in the vascular endothelium. In the present study, we disclosed that PM2.5 exposure induced the activation of the transcriptional factor AP-1 and its components, c-Jun and ATF2, in the human vascular endothelial cells. Although the DNA-binding sites for AP-1 were identified within the promoter regions of AGT, ACE and AT1R genes, RT-PCR and immunoblot assays indicated that AP-1 transactivation was only involved in AT1R upregulation, but did not affect the induction of AGT and ACE expression under the same conditions. Furthermore, ERKs and p38K functioned as the upstream protein kinases involving in AP-1 transactivation and AT1R upregulation under PM2.5 stimulation. In addition, the oxidative stress and proinflammatory responses in the PM2.5-treated vascular endothelial cells were significantly reduced when MAPKs and AP-1 activation were inhibited. Therefore, we conclude that PM2.5 exposure induces MAPK/AP-1 cascade activation, which contributes to AT1R upregulation and vascular endothelial dysfunction. Identifying novel therapeutic targets to alleviate AP-1 transactivation and restore AT1R expression may be helpful for the management of PM2.5-induced CV burden.

Telmisartan Protects a Microglia Cell Line from LPS Injury Beyond AT1 Receptor Blockade or PPARγ Activation.

The Angiotensin II Receptor Blocker (ARB) Telmisartan reduces inflammation through Angiotensin II AT1 receptor blockade and peroxisome proliferator-activated receptor gamma (PPARγ) activation. However, in a mouse microglia-like BV2 cell line, imitating primary microglia responses with high fidelity and devoid of AT1 receptor gene expression or PPARγ activation, Telmisartan reduced gene expression of pro-injury factors, enhanced that of anti-inflammatory genes, and prevented LPS-induced increase in inflammatory markers. Using global gene expression profiling and pathways analysis, we revealed that Telmisartan normalized the expression of hundreds of genes upregulated by LPS and linked with inflammation, apoptosis and neurodegenerative disorders, while downregulating the expression of genes associated with oncological, neurodegenerative and viral diseases. The PPARγ full agonist Pioglitazone had no neuroprotective effects. Surprisingly, the PPARγ antagonists GW9662 and T0070907 were neuroprotective and enhanced Telmisartan effects. GW9226 alone significantly reduced LPS toxic effects and enhanced Telmisartan neuroprotection, including downregulation of pro-inflammatory TLR2 gene expression. Telmisartan and GW9662 effects on LPS injury negatively correlated with pro-inflammatory factors and upstream regulators, including TLR2, and positively with known neuroprotective factors and upstream regulators. Gene Set Enrichment Analysis (GSEA) of the Telmisartan and GW9662 data revealed negative correlations with sets of genes associated with neurodegenerative and metabolic disorders and toxic treatments in cultured systems, while demonstrating positive correlations with gene sets associated with neuroprotection and kinase inhibition. Our results strongly suggest that novel neuroprotective effects of Telmisartan and GW9662, beyond AT1 receptor blockade or PPARγ activation, include downregulation of the TLR2 signaling pathway, findings that may have translational relevance.

Angiotensin II type 1 receptor gene polymorphism and serum angiotensin-converting enzyme level in Egyptian children with systemic lupus erythematosus.

Angiotensin II, the major effective molecule of the renin-angiotensin system, plays a vital role in the development of systemic lupus erythematosus (SLE). To study angiotensin II type 1 receptor (AT1R) gene polymorphism at (A1166C) in Egyptian children with SLE and its correlation with serum ACE level and SLE manifestations. AT1R gene polymorphism (A1166C) was done in 123 children with SLE in comparison to 100 healthy controls using polymerase chain reaction-based restriction fragment length polymorphism method (PCR-RFLP) and the tetra-primer amplification refractory mutation system-polymerase chain reaction (T-ARMS-PCR) to confirm the results of the genotyping. Serum ACE level measurement was done using ELISA technique. The frequencies of C-containing genotypes (AC + CC) and C-allele of AT1R (A1166C) were significantly higher in SLE patients compared to controls (p < 0.0001, OR = 4.9, 95% CI = 2.7-8.8; p˂0.0001, OR = 3.6, 95% CI = 2.2-5.9, respectively). Lupus nephritis (LN) patients had significantly higher frequency of (AC + CC) genotypes and C-allele compared with controls (p˂0.0001, OR = 5.1, 95% CI = 2.7-9.7; p˂0.0001, OR = 3.5, 95% CI = 2.1-6.02, respectively). Mean serum ACE levels were significantly higher in SLE patients compared to controls (p˂0.0001). There were no associations between AT1R gene polymorphism and serum ACE level and the clinical manifestations of SLE. The AT1R gene polymorphism can be considered a risk factor for the development of SLE in Egyptian children.

The expressional disorder of the renal RAS mediates nephrotic syndrome of male rat offspring induced by prenatal ethanol exposure

This study aimed to prove that prenatal ethanol exposure (PEE) can induce nephrotic syndrome in male rat offspring and to explore the underlying intrauterine programming mechanisms. Pregnant Wistar rats were intragastrically administered ethanol (4 g/kg d) from gestational day (GD) 9 to GD 20, and the male fetuses were delivered by cesarean section at GD20 and the male adult offspring were euthanized at postnatal week (PW) 24. In vitro, the primary metanephric mesenchyme cells were treated with ethanol at concentrations of 15–60 mM. The results indicated that the kidneys of adult offspring in the PEE group exhibited glomerulosclerosis as well as interstitial fibrosis. The levels of serum creatinine and urine protein were elevated; the serum total cholesterol level was increased and the serum albumin concentration was reduced. In the fetal kidney, developmental retardation was presented in the PEE group via pathological examinations, accompanied by the expressional inhibition of the glial-cell-line-derived neurotrophic factor/c-ret tyrosine kinase receptor (GDNF/c-ret) signaling pathway. Although serum angiotensin II (Ang II) level and the gene expression of renal angiotensin-converting enzyme (ACE) were increased in the PEE group, the expression of renal angiotensin II type 2 receptor (AT2R) was significantly inhibited, accompanied by a reduction in the H3K27ac level on the AT2R gene promoter. In the non-classical renin-angiotensin system (RAS), the expression of renal angiotensin converting enzyme 2 (ACE2) and Mas receptor (MasR) were inhibited in the PEE group. The above changes of the classical and non-classical RAS all sustained from utero to adulthood. In vitro, ethanol elevated the gene expression of ACE and angiotensin II type 1a receptor (AT1aR) whereas it reduced the expression of AT2R, ACE2, and MasR, accompanied by a reduction in the H3K27ac level on AT2R gene promoter. Taken together, these results suggested that PEE can induce fetal kidney developmental retardation and adult nephrotic syndrome, and direct regulation of ethanol to the renal RAS was involved in the mechanism of nephrotic syndrome induced by PEE.

Sorbitane Monostearate and Cholesterol based Niosomes for Oral Delivery of Telmisartan.

Niosomes are non-ionic surfactant vesicles used as drug carriers for encapsulating both hydrophobic and hydrophilic drugs. The aim of the present study was to prepare and characterize niosomes formulations for oral delivery of telmisartan and evaluated for its antihypertensive activity. Telmisartan loaded niosomes were prepared using thin film hydration method by varying the Span 60 and cholesterol at several molar ratios and characterized for vesicles size, polydispersity index, zeta potential, entrapment efficiency. The in vivo antihypertensive study of optimized formulation and molecular impact of angiotensin II type-1 receptor (AT1R) messenger Ribonucleic acid (mRNA) and protein expression on smooth vascular muscles of aorta was determined by real-time polymerase chain reaction (RT-PCR) and western blot analysis in Wistar albino rats. The optimized niosomes formulation NS6 presented vesicles size of 618.47 nm, polydispersity index of 0.86, with entrapment efficiency of 83.83% and possesses negative charge. In vivo study showed that the optimized formulation could reduce the systolic blood pressure in methyl prednisolone acetate induced hypertensive rats in close proximity to normal range of systolic blood pressure and maintain it over an extended period. In addition, telmisartan loaded niosomes treatment to hypertensive rats significantly attenuates the raised mRNA level and protein level of AT1R gene in comparison to hypertensive rats. Results of present study confer the potential of developed niosomes as suitable carriers for improved oral delivery of telmisartan.

Анализ ассоциаций полиморфизмов G1444A гена PPARGC1A и A1166C гена AGTR1 с преморбидными особенностями личности при шизофрении

Background: it is assumed that a number of metabolic changes and initial psychopathological anomalies of patients with endogenous psychoses are formed long before the manifestation of the disease, is associated with a hereditary, genetic component and can be reflected in the constitutional features of the patients already in the premorbid period of the disease and be a promising method for early recognition. Previously, the participation of PPARGC1A (Peroxisome proliferator-activated receptor gamma coactivator 1-alpha) and AGTR1(Angiotensin II Receptor Type 1) genes in the formation of both metabolic and morphofunctional, neurodegenerative, cognitive and other mental disorders was repeatedly demonstrated. The aim of study was to study the frequency distribution of the polymorphic markers G1444A of the PPARGC1A gene (rs8192678) and A1166C of the AGTR1 gene (rs5186) in patients with schizophrenia (ICD-10: F20, F25), depending on their afliation to the subgroups isolated according to the severity of premorbid personality anomalies: accentuated, with personality disorder (or psychopathy), with pseudopsychopathy. Material: a sample of patients was formed of in-patients (all men) of clinical unit of FSBSI «Mental health research centre». The control group consisted of 290 healthy men. DNA for the study was isolated from the venous blood of the recipients. Methods: genotyping was performed by standard PCR with further analysis of MspI restriction fragments (for rs8192678) and DstDEI fragments (for rs5186). Statistical processing of data was carried out using the program Statistica 6.0. Results: the association of studied markers with the severity of premorbid personality anomalies in patients was revealed. Thus, in the subgroup of patients with personality disorder, an increase in the frequency of the CC genotype (A1166C of the AGTR1 gene) and a decrease in the frequency of the GG genotype (G1444A of the PPARGC1A gene) compared to other subgroups; and in the subgroup of patients with pseudopsychopathy — a decrease in the frequency of the genotype CC (A1166C gene AGTR1) and an increase in the frequency of the genotype GG (G1444A gene PPARGS1A). This may indicate the possible involvement of these genes in the formation of psychopathological and biological changes in patients with schizophrenia at the stages preceding the manifestation of the disease.

AGTR1 rs5186 variants in patients with type 2 diabetes mellitus and nephropathy

Diabetic nephropathy (DN) is one of the leading causes of death in patients with type 2 diabetes mellitus (DM) and genetic factor can influence development of this complication. Association of Angiotensin II receptor type 1 (AGTR1) gene and DN has been evaluated in recent years. This study aim was to assess the association of AGTR1 gene polymorphisms with DN in a group of Iranian diabetic patients. In the present study, 97 patients with DM, 94 patients with DN and 100 healthy controls were included. Allele and genotype frequencies of AGTR1A1166C (rs5186) were determined by ARMS PCR technique. In all groups, blood glucose concentration, creatinine, urea and HbA1c were measured and urine albumin to creatinine ratio (ACR) as well as glomerular filtration rate (GFR) was calculated. A statistically significant association was found between C allele of rs5186 and DN (odds ratio (OR)=1.84 [95% CI: 1.10–3.08]; p_value=0.03). Among patients with DM, carriers of C allele (AC+CC) had significantly higher ACR (p_value=0.04) compared to AA homozygote patients. This study showed the association of AGTR1 gene polymorphisms (rs5186) with increased urinary albumin excretion in a group of Iranian patients with DM. Also, C allele can be introduced as a risk allele for DN.

Different protein kinase C isoenzymes mediate inhibition of cardiac rapidly activating delayed rectifier K+ current by different G-protein coupled receptors.

Elevated angiotensin II (Ang II) and sympathetic activity contributes to a high risk of ventricular arrhythmias in heart disease. The rapidly activating delayed rectifier K+ current (IKr ) carried by the hERG channels plays a critical role in cardiac repolarization, and decreased IKr is involved in increased cardiac arrhythmogenicity. Stimulation of α1A -adrenoreceptors or angiotensin II AT1 receptors is known to inhibit IKr via PKC. Here, we have identified the PKC isoenzymes mediating the inhibition of IKr by activation of these two different GPCRs. The whole-cell patch-clamp technique was used to record IKr in guinea pig cardiomyocytes and HEK293 cells co-transfected with hERG and α1A -adrenoreceptor or AT1 receptor genes. A broad spectrum PKC inhibitor Gö6983 (not inhibiting PKCε), a selective cPKC inhibitor Gö6976 and a PKCα-specific inhibitor peptide, blocked the inhibition of IKr by the α1A -adrenoreceptor agonist A61603. However, these inhibitors did not affect the reduction of IKr by activation of AT1 receptors, whereas the PKCε-selective inhibitor peptide did block the effect. The effects of angiotensin II and the PKCε activator peptide were inhibited in mutant hERG channels in which 17 of the 18 PKC phosphorylation sites were deleted, whereas a deletion of the N-terminus of the hERG channels selectively prevented the inhibition elicited by A61603 and the cPKC activator peptide. Our results indicated that inhibition of IKr by activation of α1A -adrenoreceptors or AT1 receptors were mediated by PKCα and PKCε isoforms respectively, through different molecular mechanisms.

BP.01.07] SEVERITY OF HEMODYNAMIC AND METABOLIC DISORDERS IN PATIENTS WITH ESSENTIAL HYPERTENSION AND TYPE 2 DIABETES DEPENDING ON GENETIC POLYMORPHISM OF AGTR1 GENE

Objective: The aim of the study was to investigate the severity of hemodynamic and metabolic disorders in patients with essential hypertension (EH) and type 2 diabetes (DM2) in A1166C polymorphism of angiotensin II receptor type 1 (AGTR1) gene in Ukrainian population. Design and method: The main group consisted of 320 patients with EH and DM2; the comparison group consisted of 90 patients with EH without DM2; the control group consisted of 31 healthy individuals. We determined carbohydrate and lipid profiles, levels of adiponectin and leptin, conducted genotyping of A1166C polymorphism of angiotensin II receptor type 1 gene. Results: It was established that 61.6% of patients with EH and DM2 and 57.8% with EH without DM2 had A/C and C/C genotypes of AGTR1. As to the spectrum of these genotypes the main group and the comparison group significantly differed from the control group (p < 0.01). The patients with EH and DM2 in presence of A/C and C/C genotypes of AGTR1 gene had significantly higher blood pressure (p < 0.001), higher levels of glucose, HbA1c, insulin, HOMA-IR, leptin (p < 0.01) and significantly lower levels of high-density lipoprotein cholesterol (p < 0.001) and adiponectin (p < 0.01) as compared to A/A genotype. The difference of adypokines levels can be explained by the influence of angiotensin II activation to the change of expression of gene encoding adypokines. However in patients with EH without DM2 polymorphism of AGTR1 gene was associated with the difference blood pressure (p < 0.001), carbohydrate (p < 0.01) and lipid (p < 0.001) spectra but did not affect adipokines levels. The absence of differences adypokines levels in the comparison group depending on AGTR1 polymorphism, unlike the main group, can indicate that the association of metabolic parameters with polymorphism of AGTR1 gene is more pronounced in presence of DM2. Conclusions: In Ukrainian population of patients with EH polymorphism of AGTR1 gene was associated with the severity of hemodynamic and metabolic disorders. In comorbidity of EH and DM2 A/C and C/C genotypes of AGTR1 gene were associated with more pronounced adypokines disbalance as compared to A/A genotype of AGTR1 gene.

Cardiovascular Protective Effects of AT2R Activation by Peptide Drug NP‐6A4

Introduction and ObjectiveLiterature shows that elevated Angiotensin II Type 2 receptor (AT2R) expression is cardiovascular reparative and mediates protection from hypertension, as well as vascular and renal injury. AT2R is a member of the anti‐inflammatory branch of the renin‐angiotensin system (RAS). Clinically, loss of AT2R expression in men due to the intronic G1675A or A1818T polymorphism is associated with impaired kidney function, pulse pressure and increased arterial stiffness. Given the pathological consequences of the loss of AT2R expression there is need for a drug that can increase AT2R gene (Agtr2) expression to mitigate cardiovascular damage. Currently there are no FDA‐approved drugs that meet these parameters. NP‐6A4 is a peptide agonist for AT2R, recently developed by Novopyxis Inc. We have shown that NP‐6A4 increases the viability of nutrient‐stressed mouse and human cardiovascular cells. Chemotherapeutic agents such as anthracyclins are inducers of cardiotoxicity. Therefore we investigated whether NP‐6A4 can protect cardiac cells from Doxorubicin (Dox; an anthracyclin) induced toxicity. To test the translational potential of a treatment paradigm that utilizes NP‐6A4 to improve cardiac functions and mitigate cardiac structural damage, we investigated the effects of NP‐6A4 treatment in Male Zucker Obese (ZO‐M) rats, a model for heart failure with preserved ejection fraction and cardiomyopathy.MethodsMTS assay (BioVision) was used to determine the effect of Dox treatment (0.5–2 μM) with and without co‐treatment with NP‐6A4 (1μM) on the survival of H9c2 cardiomyoblasts. To evaluate the effect of NP‐6A4 on cardiac structure and function, we treated 11‐week‐old ZO‐M rats that had fully developed cardiovascular disease with NP‐6A4 (1.8mg/kg/day by subcutaneous delivery). Echocardiography was used to assess cardiac function parameters. Cardiac architecture, hypertrophy, capillary density and fibrosis were assessed by staining using hematoxylin and eosin, Helix pomatia agglutinin (HPA), isolectin B4 (IB4) and Picrosirius Red (PSR) respectively. Expression levels of AT2R in the heart were assessed by quantitative RT‐PCR.Results24‐hour Dox treatment of H9c2 cardiomyoblasts suppressed cell viability (78% for 2μM vs untreated) and co‐treatment with NP‐6A4 restored cell survival. NP‐6A4 improved several cardiac parameters including endocardial circumferential strain (p≤0.05), myocardial performance index (MPI) (p≤0.005), and E/E' ratio (p≤0.002). NP‐6A4 reduced cardiomyocyte hypertrophy and fibrosis, and increased capillary density (p≤0.05). Importantly, cardiac Agtr2 (AT2R) mRNA levels were increased by about 9‐fold in NP‐6A4 treated rats vs. saline treated rats. Histopathology staining revealed regions of micro‐infarcts present in the saline group which were largely absent in the NP‐6A4 treated animals.ConclusionNP‐6A4 could mitigate Dox‐induced cardiac cell toxicity in vitro. NP‐6A4 also improved cardiac functions in a murine model of heart failure with preserved ejection fraction and cardiomyopathy, decreased risk of cardiac event (as measured by E/E'), mitigated myocardial structural damage, and improved cardiac vascularization. To our knowledge, NP‐6A4 is the first AT2R agonist that can increase cardiac AT2R expression. Based on these observations, we propose that this therapeutic candidate has the potential to treat chemotherapy‐induced cardiotoxicity as well as heart failure.Support or Funding InformationThis work was supported by NIH grant 1R01HL118376‐01 (LP), a small grant from Novopyxis Inc (LP), and Research Services facilities of HSTMVH.

Analysis of Association of Angiotensin II Type 1 Receptor Gene A1166C Gene Polymorphism with Essential Hypertension.

The A/C transversion at 1166 of the angiotensin II Type 1 Receptor (AT1R) gene per se does not characterize any functional diversity but has been associated with expression of the AT1R, consequently molecular variants of the gene may modulate the possible risk of essential hypertension. The present study was performed to determine the genotypic frequency of the A1166C polymorphism of the AT1R gene in essential hypertensive patients with the aim to assess the effect of variants of this polymorphism in hypertension. AT1R gene amplification was performed by PCR and A1166C polymorphism was determined by enzyme digestion methodologies in 224 consecutively enrolled essential hypertensive patients and 257 controls. Suitable descriptive statistics was used for different variables. Results revealed that genotype and allele distribution of the A1166C variant differed significantly in hypertensives and normotensives. Allele frequency at the A1166C position was 61%A and 39%C for control and 52%A and 48%C for patients. Observed frequencies were compatible with HWE expected frequencies in cases as well as in controls. rs5186 was found to be associated with hypertension (95% CI 1.1453-2.7932, p: 0.0106). The difference remained statistically significant after the multivariate adjustment (p<0.05), with C/C variant conferring a risk of 1.74-fold of essential hypertension. This association was confirmed by inter-genotypic variations in the mean systolic and diastolic blood pressure in patients. In conclusion, genetic variation at the AT1R gene influences the risk of hypertension stratification and might serve as a predictive marker for the susceptibility to hypertension among affected families.