Lack of association between endothelial nitric oxide synthase glu298Asp variation, visceral obesity and insulin related phenotypes in Turkish type 2 diabetic patients

Nitric oxide (NO) is an endothelium derived relaxing factor (EDRF) important in regulating heart-vessel physiology. The objective of this study was to investigate whether the eNOS gene Glu298Asp variation influenced the lipid parameters, visceral obesity, insulin related phenotypes and type 2 diabetes mellitus (T2DM) development, for the first time in a Turkish study group. We analyzed the the eNOS gene Glu298Asp genotype frequencies in 115 type 2 diabetic and 68 healthy control subjects. Serum lipids and insulin-related phenotypes were also analyzed. No significant difference for genotypic frequencies was observed for the Ban II (Eco241) restriction site in T2DM patients as compared to controls. eNOS Glu298Asp polymorphism was not found to affect visceral obesity and insulin related phenotypes. However, T2DM patients with Asp/Asp genotype were found to have lower hepatic insulin sensitivity (HIS) in comparison to Glu/Glu. In healthy controls, the insulin and HOMA levels were found to be lower in Glu/Asp genotype with respect to Glu/Glu genotype carriers (p>0.05). In T2DM patients, visceral obesity was observed in higher frequencies with Asp/Asp genotype, in comparison to Glu/Glu genotype. eNOS Glu298Asp polymorphism was not found to affect serum lipid levels in the T2DM group. However in the control group, lower serum apoB levels were observed in Asp/Asp genotype carriers in comparison to Glu/Glu genotype (p ≤ 0.05). The eNOS gene Glu298Asp polymorphism was not found to be associated with T2DM in the present study group. Although not significant, since the eNOS Glu298Asp genotypes were found to be related to HIS, insulin, HOMA and visceral obesity in the present study, further studies on larger samples are needed to explore the exact role of eNOS Glu298Asp polymorphism in insulin related phenotypes and visceral obesit

Investigation of the preventative roles of manganese superoxide dismutase Ala16Val gene polymorphism in coronary artery events

Amaç: Araştırmamızın amacı, mangan süperoksit dismutaz (MnSOD) genine ait Ala16Val polimorfizminin koroner arter hastalığı (KAH) olan kişilerdeki genotip sıklıklarını belirlemek ve genotip-kalp damar hastalığı ve genotip-biyokimyasal parametreler ve serum MnSOD aktivitesi ile etkileşimin değerlendirilmesidir. Gereç ve yöntemler: MnSOD geni Ala16Val genotipleri kantitatif polimeraz zincir reaksiyonu yöntemi ile saptandı. Serum MnSOD aktivitesi ELİSA yöntemi ile belirlendi. Bulgular: MnSOD geni Ala16Val polimorfizmi için genotip sıklıkları KAH grubunda C/C (Homozigot yabanıl tip), C/T (Heterozigot), T/T (Homozigot polimorfik tip) genotipler için sırasıyla %20.9, %41.9, %37.2 ve KAH olmayan grupta %15.5, %51.7, %32.8 olarak tespit edildi. Tüm çalışma grubundaki mitokondriyal MnSOD aktiviteleri, MnSOD geni Ala16Val genotiplerine göre karşılaştırıldığında CC genotip taşıyıcılarının en yüksek, TT genotip taşıyıcılarının en düşük MnSOD aktivitesine sahip olduğu belirlendi. Koroner arter hastalığı bulunmayan hastalarda MnSOD Ala16Val polimorfizminin yağsız vücut kütlesi (p=0.01) ve yağ kütlesi (p=0.05) üzerine anlamlı etkileri olduğu saptandı. Ala16Val polimorfizminin heterozigot genotipe sahip KAH hastalarında yağdan zengin gıdalarla beslenme alışkanlıkları diğer genotiplere kıyasla daha düşük iken, homozigot polimorfik genotipe sahip KAH hastalarının diğer genotiplere kıyasla proteinden fakir beslenme alışkanlıklarını benimsedikleri görüldü. Sonuç: Tüm çalışma grubu değerlendirildiğinde, MnSOD Ala16Val genotipleri ile beslenme alışkanlıkları arasında anlamlı bir ilişki bulunurken yağsız vücut kütlesi üzerinde etkili bulunmamıştır.Objectives: The aim of this study was to determine the genotypic frequencies of manganese superoxide dismutase (MnSOD) gene Ala16Val polymorphism frequencies and to evaluate the effects of MnSOD Ala16Val variation over coronary artery disease (CAD) in which people, biochemical parameters and MnSOD activity. Materials and methods: MnSOD gene Ala16Val polymorphisms were determined with quantitative polymerase chain reaction method. MnSOD activity in serum were determined by ELISA method. Results: The MnSOD gene Ala16Val genotype frequencies were determined respectively as 20.9%, 41.9%, 37.2% for homozygous wild genotype (C/C), heterozygous genotypes (C/T), homozygous polymorphic genotype (T/T) in the CAD patients; whereas 15.5%, 51.7% and 32.8% for the patients without CAD. In total study group, C/C genotype carriers were detected to have the highest, whereas T/T genotype carriers were detected to have the lowest mitochondrial MnSOD activities when compared according to MnSOD gene Ala16Val genotypes. In the patients without CAD, MnSOD Ala16Val polymorphism was found to have significant effects on lean body mass (p=0.01) and fat mass (p=0.05). In CAD patients with Ala16Val polymorphism heterozygous genotype oil-rich eating habits were lower to that of other genotypes, whereas CAD patients with homozygous polymorphic genotype was detected to prefer protein-poor diet. Conclusion: When all the study group was evaluated, MnSOD Ala16Val genotypes were not found to have statistically significant relationship with eating habits, but found to be effective only on lean body mass

Omentum adiposity is linked with resistin gene expression

Background: This study demonstrated site-specific adipose tissue resistin gene expression differences in individuals with and without type 2 diabetes mellitus. The relationship between conventional drug therapy and adipose tissue resistin gene expression was also determined. Paired omental and subcutaneous adipose tissues were excised during elective surgery from morbidly obese and obese patients. Methods: Resistin mRNA expressions were determined by qPCR. All tissue sections also were also analyzed for their resistin and CD68 protein expressions by immunohistochemistry. Results: No significant difference for omental and subcutaneous adipose tissue resistin mRNA expression levels were found among morbidly obese and obese study groups. The omental adipocytes resistin mRNA expressions increased with macrophage number both in the omental and subcutaneus fat. Resistin mRNA expressions of the omental and subcutaneous fat were in positive correlation. As the omental adipocytes radius decreased, the macrophage number increased in subcutaneous fat. In the omentum the adipocytes diameter and areas increased, in correlation with macrophage number. The antidiabetic drug use was found to increase adipocyte size both in the omentum and subcutaneous fat. Conclusions: The higher resistin gene expression in the omental fat may induce the increase in size and number of adipocytes, thus leading to elavation in omental fat mass

Alterations in niban gene expression as a response to stress conditions in 3T3-L1 adipocytes

Adipocyte death is important in obesity development. Understanding and prevention of adipocyte deaths may be a molecular approach in the treatment. In the study, we aimed to understand role of Niban gene, which acts as an anti-apoptotic molecule as a response to stress conditions, in adipocytes. 3T3-L1 adipocytes were treated with different doses of linoleic acid, hydrogen peroxide and ethanol; and proliferation of the cells examined with real time monitoring iCELLingence system. Gene expression levels were measured by q-PCR. As a response to 24h 480 mu M linoleic acid treatment, Niban gene expression was found to be higher than control group (p = 0.008), whereas 24 h 90 mM ethanol treatment was determined to be lower than control group (p = 0.008). The highest value of Niban gene expression among H2O2 treatment groups was detected in 4h 600 mu M H2O2 in comparison to control group (p = 0.008). To understand role of Niban in adipogenesis, Niban gene expressions were compared between pre-adipocytes and advanced fat accumulated adipocytes and determined to be significantly different (p = 0.042). Our results suggest that Niban might be involved in stress response process in adipocytes. However, the exact molecular role of Niban needs to be investigated in further studies

The effects of glipizide on DNA damage and nuclear transport in differentiated 3T3-L1 adipocytes

Background Despite commonly use for treatment of type II diabetes, possible effects of glipizide on nuclear transport and DNA damage in cells are unknown. Since clinical response of glipizide may change with aging, the aim of the study was to investigate the effect of glipizide by comparing mature and senescent adipocytes. Methods and results The effects of glipizide were investigated in 3T3-L1 adipocytes. Effective and lethal doses were determined by real-time monitoring iCELLigence system. Comet assay was performed to determine DNA damage and quantitative PCR was conducted to detect gene expression levels. RAN expressions were found to be up regulated in mature 180 mu M glipizide treated adipocytes compared to control group (p < 0.05); whereas down regulated in senescent 180 mu M glipizide treated adipocytes compared to their control adipocytes (p < 0.05). Olive Tail Moment values were significantly higher in mature 180 mu M glipizide treated adipocytes (MTG) and senescent 180 mu M glipizide treated adipocytes (STG) comparing their untreated controls (p < 0.001 and p < 0.001 respectively). Also class 5 comets that shows severe DNA damage were found to be higher in both MTG and STG groups than their controls (p < 0.001 and p < 0.001, respectively). OTM values were higher in STG than MTG (p < 0.001). Conclusions This is the first study that reports glipizide caused DNA damage increasing with senescence in adipocytes. As a response to glipizide treatment Ran gene expression increased in mature; and decreased in senescent adipocytes. Further studies are needed to reveal the effect of glipizide on DNA and nuclear interactions in molecular level

The effects of SNAI1 rs6125849 gene polymorphism on metastasis and survival in colorectal cancer: Preliminary results from Turkish subjects

Background: Since metastasis is one of the leading causes of high mortality in colorectal cancer, uncovering the molecular mechanisms underlying metastatic process has become important to find new treatment approaches. With this perspective, in this study we investigated effects of epithelial-mesenchymal transition (EMT) inducer SNAI1 rs6125849 gene polymorphism on different features of colorectal cancer. Material and methods: A total of 141 Turkish subjects consisting of 72 colorectal cancer patients and 69 healthy controls were included in the study. SNAI1 rs6125849 genotype analyses were performed with Agena MassARRAY platform. Results: We did not find any significant difference between case and control groups for rs6125849 genotypes. All metastatic patients were detected to have homozygous mutant (AA) genotype. Heterozygous and homozygous mutant genotypes (GA + AA) were more common in metastatic colorectal cancer patients compared to those with homozygous wild type (GG) genotype (p = 0.066). We also detected positive, albeit low but significant correlation between rs6125849 genotypes in codominant and dominant models (r = 0.250, p = 0.034 and r = 0.234, p = 0.048, respectively). Overall survival times was found to be considerably higher patients with A allele (Log rank: 3.04, p = 0.081). Conclusion: According to our preliminary results, we may speculate that SNAI1 gene G > A variation may result with a metastatic phenotype and shorter overall survival. Due to limited number of patients in our study group, further in vitro and in vivo studies are required to understand the role of SNAI1 rs6125849 variation on metastasis and survival functions

DNA Damage in AML-12 Hepatocytes and 3T3-L1 Adipocytes Treated with Clopidogrel

Background: Clopidogrel has been commonly prescribed as a selective P2Y(12) receptor antagonist to reduce heart attack and stroke risk. Nearly 10% of absorbed clopidogrel is metabolized to active forms by cytochrome P450 (CYP) enzymes in the liver and 90% to inactive clopidogrel carboxylate by esterases. Objective: Since different forms of clopidogrel have cytotoxic potential, our aim was to determine the effect of 7.5, 40, and 75 mu M clopidogrel over DNA damage in adipocytes and hepatocytes. Methods: In the present study, DNA damage was investigated by Comet analysis using 3T3-L1 adipocytes and Alpha Mouse 12 (AML-12) hepatocytes. Results: DNA fragmentation was found to be increased as a response to 7.5 mu M, 40 mu M, and 75 mu M clopidogrel treatment compared to non-treated control groups in AML-12 hepatocytes (p<0.01, p<0.001, p<0.01 respectively) and 3T3-L1 adipocytes (p<0.001, p<0.001 and p<0.001respectively). DNA damage levels as a response to clopidogrel treatment were found to be higher in 3T3-L1 adipocytes than AML-12 hepatocytes. Also, DNA damage levels in adipocytes and hepatocytes were found to increase dose-dependently for 7.5 and 40 mu M clopidogrel, whereas decreased as a response to 75 mu M. Conclusion: According to our results, clopidogrel results in more DNA damage in adipocytes than in hepatocytes. The molecular mechanism of clopidogrel genotoxicity needs to be further investigated especially in adipose tissue

Association of PD-1 and PDL-1 gene polymorphisms with colorectal cancer risk and prognosis

Background Programmed Cell Death-1 (PD-1) together with Programmed Death Ligand 1 (PDL-1) have crucial roles in anti-tumor immune response, cancer susceptibility and prognosis. Since PD-1 and PDL-1 have been considered as important genetic risk factors in cancer development and their functions can be affected by polymorphic sites, we investigated the effects of PD-1 rs2227981, rs2227982, rs36084323 and PDL-1 rs2282055, rs822336 gene polymorphisms on colorectal cancer (CRC) risk and prognosis in Turkish subjects. Methods and results Our study group consisted of 5-FU or Capacitabine prescribed CRC diagnosed patients and healthy controls. Genotype analyses of PD1 and PDL-1 polymorphisms were performed with Agena MassARRAY platform. rs36084323 CT genotype frequency was found to be higher in controls compared to cases (p C polymorphism might be useful in predicting CRC prognosis. PDL-1 rs2282055 T > G polymorphism might be useful in predicting both CRC risk and prognosis. Further studies should be conducted in larger and different populations to clear the roles of PD-1 and PDL-1 polymorphisms in CRC risk and prognosis