Razi Journal of Medical Sciences

fa پلی‌مورفیسم TMPRSS6rs2111833 در مبتلایان به فقر آهن؛ یک مطالعه مورد – شاهدی TMPRSS6rs2111833 Polymorphism in Iron Deficiency Patients; A Case-Control Study ژنتیک Genetic پژوهشي Research <div style="text-align: justify;"> زمینه و هدف: آهن یک ماده معدنی ضروری برای زندگی است و تقریباً هر سازمان زنده‌ای به آهن نیاز دارد. فقر آهن شایع‌ترین کم خونی در سراسر جهان با پیامدهای مهم بالینی است. این بیماری ناشی از در دسترس نبودن آهن کافی برای تولید هموگلوبین به دلایلی از جمله فقر آهن در رژیم غذایی ،جذب ناکافی آهن، خونریزی مزمن و عوامل ژنتیکی است. چندین پلی مورفیسم ژنتیکی مرتبط با وضعیت آهن با استفاده از مطالعات مرتبط با ژنوم شناسایی شده است. هدف از این مطالعه بررسی پلی‌مورفیسم‌های TMPRSS6rs2111833 در مبتلایان به فقرآهن در غرب ایران است. روش کار: تحقیق حاضر یک مطالعه مورد – شاهدی است. نمونه‌گیری در یک فاصله زمانی 3 ماهه در زمستان 1399 و با همکاری آزمایشگاه‌های خصوصی در سطح شهر سنندح، دهگلان و کرمانشاه انجام شد. در مجموع 91 نفر با رضایت شخصی انتخاب شده و در این مطالعه قرار گرفتند. از این تعداد براساس نتایج فریتین و همچنین توجه به پارامترهای RBC، WBC‌ها و پلاکت‌ها  43 نفر افرادی با فقر آهن و 48 نفر کنترل سالم انتخاب شدند. از هر شرکت کننده 5 میلی لیتر خون اخذ شد و به لوله‌های CBC حاوی ماده ضد انعقاد انتقال یافت. بعد از استخراج DNA؛ فراوانی آللی پلی مورفیسم مورد نظر افراد با استفاده از تکنیک ARMS-PCR بررسی شد. در انتها آزمون آماری برای مقایسه فراوانی آللی در نمونه‌های شاهد و افرادی با فقر آهن با استفاده از ابزار SPSS ویندوز انجام شد و مقدار 05/0 p < از نظر آماری توصیف شد. یافته‌ها: افرادی با فقر آهن 25/47 و افراد سالم 75/52 درصد افراد مورد بررسی را تشکیل دادند. در مجموع  فراوانی GG، AG و AA  در کل شرکت کنندگان بترتیب 45، 22 و 33 درصد بود. از این میزان سهم افرادی با فقر آهن و سالم برای 3 ترادف آللی به ترتیب47 و 44 درصد،  23 و 21 درصد و 30 و 35 درصد بود. همچنین از مجموع 182 آلل، 102 آلل معادل 56 درصد آلل‌ها G و 80 آلل معادل 44 درصد آلل A بود. سهم آلل G در افرادی با فقر آهن و گروه کنترل به ترتیب 58 و 54 درصد و سهم آلل A 42 و 46 درصد بود. همچنین آزمون هاردی-وینبرگ نشان داد که جمعیت مورد ارزیابی در این تجزیه و تحلیل متعادل است. تمام مدل‌های ممکن هم بارز، بارز، مغلوب و فوق بارز بررسی شد و در تمام موارد میزان Pv بزرگ‌تر از  05/0 بود که نشان از عدم وابستگی این پلی مورفیسم با  فقر آهن در جمعیت مورد بررسی داشت. نتیجه‌گیری: پلی مورفیسم rs2111833 ارتباط معنی‌داری با فقر آهن در جمعیت مورد بررسی نشان نداد. علاوه بر این، هیچ ارتباط معنی‌داری بین این پلی مورفیسم و پارامترهای بالینی هموگلوبین، آهن، فریتین، RBC‌ها، WBC‌ها و پلاکت‌ها مشاهده نشد.</div> <div style="text-align: justify;">Background & Aims: Iron deficiency anemia is the most common anemia worldwide with significant clinical consequences. The disease is caused by not having enough iron to produce hemoglobin for reasons such as dietary iron deficiency, insufficient iron absorption, chronic bleeding, and genetic factors. Hepcidin protein is the main regulator of systemic iron homeostasis hormone ferroportin. The role of hepcidin is to control the expression of iron-releasing ferroportin levels in duodenal cells and macrophages, modulating iron uptake and recycling. Hepcidin inactivation causes severe iron overload, while increased hepcidin causes anemia. Several factors regulate hepcidin synthesis, including matriptase-2, which reduces hepcidin synthesis. Matryptase-2 is a type 2 membrane serum proteinase, encoded by the TMPRSS 6 gene. This gene is located on the long arm of chromosome 22 and is mainly expressed in the liver. In humans, inactivation of this protein leads to a rare autosomal recessive disease called IRIDA. Several genetic polymorphisms related to iron status have been identified using genome-related studies. The aim of this study was to investigate the polymorphism of TMPRSS6rs2111833 in patients with iron deficiency anemia in the west of the country. Methods: The present study is a case-control study. Using the sample size determination method available, sampling was performed at a time interval of 3 months with the cooperation of private laboratories in Sanandaj, Dehgolan and Kermanshah. A total of 91 people were randomly included with respect personal consent. Of these, according to the results of ferritin, hemoglobin, red blood cells, white blood cells and platelets 43(47.25%) with iron deficiency anemia and 48(52.75%) healthy controls were enrolled. Five ml of blood sample was collected from all participants and placed in CBC tubes containing anticoagulant and after inverting several times, it was placed in freezer -20. DNA extraction was performed using the extraction kit of Kwsar Company and according to the kit instructions. The quantity and quality of the extracted DNAs were determined using light absorption in spectrophotometer and electrophoresis on 1% agarose gel. The isolated DNA was stored in separate microtubes at -20 ° C until PCR. Then the genotype of individuals was obtained using ARMS-PCR technique and primer sequences used in previous studies. Amplification of the desired fragment was performed using Sinagene Company PCR kit by Rotor-Gene Q (Corbett Life Science) cyclotron device according to the protocol and the PCR product was loaded on 1.8% agarose gel and the quantity and quality were determined. Finally, a statistical test was performed to compare the frequency of genotypes in all samples using Windows SPSS tool and the value of p <0.05 was statistically described. Results: The results showed that the frequency of GG, AG and AA in all participants were 45%, 22% and 33%, respectively, and so for sick and healthy individuals were 47 and 44, 23 and 21 and 30 and 35, respectively. Also, out of 200 alleles, 120(56%) and 80(44%) alleles were G and A, respectively. The portion of G allele in patients and controls group was 58 and 54% and the share of A allele was 42 and 46%, respectively. Hardy Weinberg and heterozygosity equilibria were also examined for the two groups. The Hardy-Weinberg test showed that the assessed population was balanced. Heterozygosity for a gene locus is defined as the frequency of heterozygous individuals for that locus relative to the total population. A gene locus is polymorphic if its heterozygosity is greater than 0.1, and if it is greater than 0.7, it is highly polymorphic. Based on the results of this study, it was seen that the difference between observed and expected heterozygosity in the studied polymorphism is less than 0.1, so the gene position is not polymorphic. Also, co-dominant, dominant, recessive and super-dominant models were examined and in all cases, the Pv value was greater than 0.05. On the other hand, Sixty-nine percent of participants had normal hemoglobin, 41% normal iron, 51% normal ferritin, 83% normal red blood cells, 92% normal platelets and 82% normal white blood cells. On the other hand, 22% of participants had abnormal hemoglobin, 65% abnormal iron, 54% abnormal ferritin, 22% abnormal red blood cells, 2% abnormal platelets and 26% abnormal white blood cells. rs 2111833 showed no significant association with decreased serum hemoglobin, serum iron, serum ferritin, platelets, red and white blood cells. Conclusion: This results showed that the rs2111833 polymorphism were not significantly associated iron deficiency anemia in the studied population. In addition, no significant association was found between rs2111833 and hemoglobin, iron, ferritin, platelets, RBCs and WBCs clinical parameters. Beutler and colleagues investigated the role of TMPRSS6 gene polymorphisms in adults with iron deficiency anemia. Their observations highlighted the role of matriptase-2 in controlling iron metabolism and erythrocyte parameters. Sylvester et al. Reported the association of TMPRSS6 mutations with iron deficiency anemia. Previously, Feinberg et al. reported several mononucleotide polymorphisms associated with iron deficiency anemia, including rs 869320724, rs 767094129, rs 786205059, rs 137853120 and rs 137853119.7  Several studies have shown that the polymorphism rs 2111833 is associated with decreased iron and hemoglobin levels therefore, it related with an increased risk in iron deficiency anemia.  In 2015, Gichohi-Wainaina et al. Identified some changes in the partial frequencies of the rs 2111833 alleles in Asian populations compared to the Caucasian population, which matched our findings. Anemia remains a widespread and significant global health problem that needs to be adequately addressed. Although iron deficiency anemia is the leading cause of anemia in most areas, recent research shows that the cause of anemia is complex and specific to each region. To better understand how to help the underlying causes of anemia, including iron deficiency anemia and other nutritional deficiencies, diseases, and hemoglobin disorders, efforts need to be made to make appropriate interventions work under certain conditions. Further studies on a larger patient scale are also necessary to identify potential haplotypes and polymorphisms responsible for the low response to oral iron therapy and may be useful for planning a proper iron supplementation. There were some limitations to this study, such as an insufficient total number of participants. However, these results could enhance our understanding of the role of genetics, particularly single nucleotide polymorphisms, in increasing susceptibility to a variety of diseases.</div> فقر آهن, کم خونی,TMPRSS6, rs2111833 Iron Deficiency, Anemia, TMPRSS6, rs2111833 183 193 http://rjms.iums.ac.ir/browse.php?a_code=A-10-2112-6&slc_lang=fa&sid=1 Mahsa Amiri Hosseini مهسا امیری حسینی 3900319475328460074327 3900319475328460074327 No MSc, Department of Biology, Sanandaj Branch, Islamic Azad University, Sanandaj, Iran کارشناس ارشد بیوشیمی، گروه زیست شناسی، واحد سنندج، دانشگاه آزاد اسلامی، سنندج، ایران Fatemeh Keshavarzi فاطمه کشاورزی gol.keshavarzi@gmail.com 3900319475328460074326 3900319475328460074326 Yes Associate Professor, Department of Biology, Sanandaj Branch, Islamic Azad University, Sanandaj, Iran دانشیار، گروه زیست شناسی، واحد سنندج، دانشگاه آزاد اسلامی، سنندج، ایران Nahid Haghnazari ناهید حق نظری 3900319475328460074328 3900319475328460074328 No Assistant Professor, Department of Biology, Sanandaj Branch, Islamic Azad University, Sanandaj, Iran استادیار، گروه زیست شناسی، واحد سنندج، دانشگاه آزاد اسلامی، سنندج، ایران