Alireza Azani1, Asghar Hosseinzadeh2, Roya Azadkhah2, Ali Akbar Poursadegh Zonouzi3, Ahmad Poursadegh Zonouzi4, Younes Aftabi5, Hourieh Khani6, Leida Heidary6, Shahla Danaii6, Nasrin Bargahi4, Nasser Pouladi7, Sayed Mostafa Hosseini8. 1. Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran. 2. Department of Biology, Tabriz Branch, Islamic Azad University, Tabriz, Iran. 3. Department of Genetics and Medical Biotechnology, Faculty of Medicine, Hormozgan University of Medical Sciences, Bandar Abbas, Iran. 4. Biotechnology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran. 5. Drug Applied Research Center, Tabriz University of Medical Sciences, Tabriz, Iran. 6. Laboratory of Medical Genetic, ART and Stem Cell Research Center (ACECR), Tabriz, Iran. 7. Department of Cellular and Molecular Biology, Azarbaijan Shahid Madani University, Tabriz, Iran. 8. Human Genetic Research Center, Baqiyatallah university of Medical Science, Tehran, Iran. Electronic address: smhosseini@bmsu.ac.ir.
Abstract
OBJECTIVE(S): Many lines of evidence suggest that reduced production of nitric oxide (NO) due to single nucleotide polymorphisms in endothelial nitric oxide synthase (eNOS) gene may affect the implantation and maintenance of pregnancy. Accordingly, our objective was to investigate whether the eNOS polymorphisms (-786 T>C, intron 4 b/a VNTR and 894 G>T) and haplotypes may be associated with increased susceptibility to recurrent pregnancy loss (RPL). STUDY DESIGN: A total of 130 women with a history of two or more unexplained consecutive first trimester miscarriages and 110 ethnically matched women with at least two normal pregnancies and no history of pregnancy loss were included in the study as cases and controls, respectively. To identify the genotypes, we used polymerase chain reaction (PCR) and PCR-restriction fragment length polymorphism (PCR-RFLP) methods In addition, an in silico analysis was conducted to predict the possible effects of the eNOS 894 G>T polymorphism on the structure and function of eNOS mRNA and protein using prediction servers. RESULTS: Our findings revealed that the prevalence of eNOS -786 T>C polymorphism, eNOS -786C allele and TC+CC genotype in cases were significantly higher than those in healthy controls (p<0.05). Also, the combination genotypes -786TT/4b4a and -786TT/894GG were significantly associated with reduced risk of RPL. We also found that the C-4a-G haplotype of the eNOS gene studied polymorphisms was significantly associated with a predisposition to RPL (odds ratio, 3.219; 95% confidence interval, 1.649-6.282; p=0.0003). The in silico analysis showed that the eNOS 894 G>T polymorphism couldn't affects eNOS mRNA and protein significantly. CONCLUSION: Our findings provide evidence to support the hypothesis that eNOS -786 T>C polymorphism and the -786C-4a-894G haplotype are associated with the high risk of RPL.
OBJECTIVE(S): Many lines of evidence suggest that reduced production of nitric oxide (NO) due to single nucleotide polymorphisms in endothelial nitric oxide synthase (eNOS) gene may affect the implantation and maintenance of pregnancy. Accordingly, our objective was to investigate whether the eNOS polymorphisms (-786 T>C, intron 4 b/a VNTR and 894 G>T) and haplotypes may be associated with increased susceptibility to recurrent pregnancy loss (RPL). STUDY DESIGN: A total of 130 women with a history of two or more unexplained consecutive first trimester miscarriages and 110 ethnically matched women with at least two normal pregnancies and no history of pregnancy loss were included in the study as cases and controls, respectively. To identify the genotypes, we used polymerase chain reaction (PCR) and PCR-restriction fragment length polymorphism (PCR-RFLP) methods In addition, an in silico analysis was conducted to predict the possible effects of the eNOS 894 G>T polymorphism on the structure and function of eNOS mRNA and protein using prediction servers. RESULTS: Our findings revealed that the prevalence of eNOS -786 T>C polymorphism, eNOS -786C allele and TC+CC genotype in cases were significantly higher than those in healthy controls (p<0.05). Also, the combination genotypes -786TT/4b4a and -786TT/894GG were significantly associated with reduced risk of RPL. We also found that the C-4a-G haplotype of the eNOS gene studied polymorphisms was significantly associated with a predisposition to RPL (odds ratio, 3.219; 95% confidence interval, 1.649-6.282; p=0.0003). The in silico analysis showed that the eNOS 894 G>T polymorphism couldn't affects eNOS mRNA and protein significantly. CONCLUSION: Our findings provide evidence to support the hypothesis that eNOS -786 T>C polymorphism and the -786C-4a-894G haplotype are associated with the high risk of RPL.
Authors: Yazeed A Al-Sheikh; Hazem K Ghneim; Adel F Alharbi; Mashael M Alshebly; Feda S Aljaser; Mourad A M Aboul-Soud Journal: Exp Ther Med Date: 2019-10-07 Impact factor: 2.447