fa بررسی متیلاسیون پروموتر ژن TNF-𝛼 در بیماران مبتلا به دیابت نوع دو عمومى General پژوهشي Research <strong><span style="font-family:b lotus;"><span style="font-size:12.0pt;">مقدمه</span></span></strong><span style="font-family:b lotus;"><span style="font-size:12.0pt;">: دیابت نوع دو شرایط التهابی مزمنی است که آمیزه‌ای از عوامل ژنتیکی و محیطی در آن نقش دارد. فاکتور نکروز توموری آلفا یا </span></span><span dir="LTR"><span style="font-family:times new roman,serif;"><span style="font-size:10.0pt;">TNF- &alpha;</span></span></span><span style="font-family:b lotus;"><span style="font-size:12.0pt;"> به‌عنوان یک سایتوکاین پیش التهابی مترشحه از بافت چربی با تأثیر بر مسیر سیگنالینگ انسولین می‌تواند در ایجاد مقاومت به انسولین در بیماران دیابت نوع دو نقش داشته باشد. با توجه اهمیت تغییرات اپی ژنتیک در ایجاد بیماری‌های چند عاملی، این مطالعه با هدف بررسی متیلاسیون پروموتر ژن </span></span><em><span dir="LTR"><span style="font-family:times new roman,serif;"><span style="font-size:10.0pt;">TNF-&alpha;</span></span></span></em><span style="font-family:b lotus;"><span style="font-size:12.0pt;"> در بیماران مبتلا به دیابت نوع دو و مقایسه‌ی آن با افراد غیر دیابتی انجام گردید. </span></span><br> <strong><span style="font-family:b lotus;"><span style="font-size:12.0pt;">روش‌ها: </span></span></strong><span style="font-family:b lotus;"><span style="font-size:12.0pt;">این مطالعه بر روی 61 بیمار دیابت نوع دو و 31 فرد غیر دیابتی انجام شد. بیماران </span></span><span style="font-family:b lotus;"><span style="font-size:12.0pt;">شامل دو </span></span><span style="font-family:b lotus;"><span style="font-size:12.0pt;">گروه مبتلایان کمتر از 5 سال و مبتلایان بیشتر از 5 سال بودند. گروه‌ها از نظر ویژگی‌های دموگرافیک همسان بودند. پروفایل لیپیدی </span></span><span style="font-family:b lotus;"><span style="font-size:12.0pt;">با کیت‌های استاندارد </span></span><span style="font-family:b lotus;"><span style="font-size:12.0pt;">اندازه‌گیری شد. سپس ناحیه‌ی پروموتر ژن </span></span><span dir="LTR"><span style="font-family:times new roman,serif;"><span style="font-size:10.0pt;">TNF-ɑ</span></span></span><span style="font-family:b lotus;"><span style="font-size:12.0pt;"> به روش تیمار با بیسولفیت، انجام </span></span><span dir="LTR"><span style="font-family:times new roman,serif;"><span style="font-size:10.0pt;">Nested PCR</span></span></span><span style="font-family:b lotus;"><span style="font-size:12.0pt;"> و نهایتاً تعین توالی بررسی گردید.</span></span><br> <strong><span style="font-family:b lotus;"><span style="font-size:12.0pt;">یافته‌ها</span></span></strong><span style="font-family:b lotus;"><span style="font-size:12.0pt;">: در مطالعه‌ی ما ارتباطی بین متیلاسون مناطق </span></span><span dir="LTR"><span style="font-family:times new roman,serif;"><span style="font-size:10.0pt;">CpG</span></span></span><span style="font-family:b lotus;"><span style="font-size:12.0pt;"> پروموتر ژن </span></span><em><span dir="LTR"><span style="font-family:times new roman,serif;"><span style="font-size:10.0pt;">TNF-ɑ</span></span></span></em><span style="font-family:b lotus;"><span style="font-size:12.0pt;"> در گروه‌های مورد مطالعه مشاهده نشد. همچنین بین زنان و مردان در ارتباط با متیلاسیون پروموتر ژن </span></span><em><span dir="LTR"><span style="font-family:times new roman,serif;"><span style="font-size:10.0pt;">TNF-ɑ</span></span></span></em><span style="font-family:b lotus;"><span style="font-size:12.0pt;"> درگروههای دیابتی و غیر دیابتی تغییری مشاهده نگردید. </span></span><br> <strong><span style="font-family:b lotus;"><span style="font-size:12.0pt;">نتیجه‌گیری: </span></span></strong><span style="font-family:b lotus;"><span style="font-size:12.0pt;">به‌نظر می‌رسد تغییرات اپی ژنتیکی سایتوکاین‌هایی که در ایجاد مقاومت به انسولین در بیماران دیابتی نوع دو نقش دارند در نمونه‌ی خون محیطی </span></span><span style="font-family:b lotus;"><span style="font-size:12.0pt;">محسوس نیست </span></span><span style="font-family:b lotus;"><span style="font-size:12.0pt;">و در این رابطه احتمالاً بافت‌های دیگر باید مورد بررسی قرار گیرند. </span></span> <strong>Background</strong>: Type II diabetes is a chronic inflammatory condition that is associated with a combination of genetic and environmental factors. Tumor necrosis factor alpha or TNF-&alpha; as an adipocyte cytokine, which affects the signaling pathway of insulin, can contribute to insulin resistance in type 2 diabetes patients. Considering the importance of epigenetic changes in multifactorial diseases, this study aimed to investigate TNF-&alpha; promoter methylation in patients with type 2 diabetes.<br> <strong>Methods</strong>: This study was performed on 61 patients with type 2 diabetes and 31 non-diabetic patients. The Groups were matched in terms of demographic characteristics. The lipid profiles were measured by standard kits. TNF-&alpha; promoter methylation levels were measured by bisulphite treatment method, Nested PCR and sequencing.<br> <strong>Results</strong>: There was no association between TNF-&alpha; promoter methylation gene promoter and type 2 diabetes in the studied groups. Also, there was no association between TNF-&alpha; promoter methylation in diabetic and non-diabetic groups between males and females.<br> <strong>Conclusion: </strong>The epigenetic changes in cytokines that contribute to insulin resistance in type 2 diabetic patients seem to be ineffective in peripheral blood samples, and other tissues may need to be investigated in this regard. دیابت نوع دو, اپی ژنتیک, TNF-α, متیلاسیونDNA Type 2 diabetes mellitus, Epigenetic, TNF-α, DNA methylation 44 50 http://ijdld.tums.ac.ir/browse.php?a_code=A-10-1133-1&slc_lang=fa&sid=1 Zahra Moradi زهرا مرادی No Wu Y, Ding Y, Tanaka Y, Zhang W. Risk factors contributing to type 2 diabetes and recent advances in the treatment and prevention. Int J Med Sci 2014; 11(11):1185-200. 2. Zheng Y, Ley SH, Hu FB. Global aetiology and epidemiology of type 2 diabetes mellitus and its complications. Nature Reviews Endocrinology [Review Article]. 2017; 14:88. 3. Kameswaran V, Bramswig NC, McKenna LB, Penn M, Schug J, Hand NJ, et al. Epigenetic regulation of the DLK1-MEG3 microRNA cluster in human type 2 diabetic islets. Cell Metab 2014; 19(1):135-45. 4. Kwak SH, Park KS. Recent progress in genetic and epigenetic research on type 2 diabetes. Exp Mol Med 2016; 48:e220. 5. Nilsson E, Ling C. DNA methylation links genetics, fetal environment, and an unhealthy lifestyle to the development of type 2 diabetes. Clin Epigenetics 2017; 9:105. 6. Makki K, Froguel P, Wolowczuk I. Adipose tissue in obesity-related inflammation and insulin resistance: cells, cytokines, and chemokines. ISRN Inflamm 2013; 2013:139239 7. Kahn SE, Hull RL, Utzschneider KM. Mechanisms linking obesity to insulin resistance and type 2 diabetes. Nature 2006; 444(7121):840-6. 8. Nicholas DA, Andrieu G, Strissel KJ, Nikolajczyk BS, Denis GV. BET bromodomain proteins and epigenetic regulation of inflammation: implications for type 2 diabetes and breast cancer. Cell Mol Life Sci 2017; 74(2):231-43. 9. Sommese L, Zullo A, Mancini FP, Fabbricini R, Soricelli A, Napoli C. Clinical relevance of epigenetics in the onset and management of type 2 diabetes mellitus. Epigenetics 2017; 12(6):401-15. 10. Al Muftah WA, Al-Shafai M, Zaghlool SB, Visconti A, Tsai P-C, Kumar P, et al. Epigenetic associations of type 2 diabetes and BMI in an Arab population. Clinical epigenetics 2016; 8: 13. 11. Akash MSH, Rehman K, Liaqat A. Tumor Necrosis Factor-Alpha: Role in Development of Insulin Resistance and Pathogenesis of Type 2 Diabetes Mellitus. J Cell Biochem 2018; 119(1):105-10. 12. Miao F, Gonzalo IG, Lanting L, Natarajan R. In vivo chromatin remodeling events leading to inflammatory gene transcription under diabetic conditions. J Biol Chem 2004; 279(17):18091-7. 13. Hermsdorff HH, Mansego ML, Campión J, Milagro FI, Zulet MA, Martínez JA. TNF-alpha promoter methylation in peripheral white blood cells: Relationship with circulating TNFα, truncal fat and n-6 PUFA intake in young women. Cytokine 2013; 64(1):265-71. 14. Liu C, Feng X, Li Q, Wang Y, Li Q, Hua M. Adiponectin, TNF-alpha and inflammatory cytokines and risk of type 2 diabetes: A systematic review and meta-analysis. Cytokine 2016; 86:100-9. 15. Pinzon-Cortes JA, Perna-Chaux A, Rojas-Villamizar NS, Diaz-Basabe A, Polania-Villanueva DC, Jacome MF, et al. Effect of diabetes status and hyperglycemia on global DNA methylation and hydroxymethylation. Endocr Connect 2017; 6(8):708-25 مرکز تحقیقات سلولی و مولکولی، پژوهشکده‌ی علوم پایه‌ی سلامت Mehrnaz Sadat Ravari مهرناز السادات راوری No Wu Y, Ding Y, Tanaka Y, Zhang W. Risk factors contributing to type 2 diabetes and recent advances in the treatment and prevention. Int J Med Sci 2014; 11(11):1185-200. 2. Zheng Y, Ley SH, Hu FB. Global aetiology and epidemiology of type 2 diabetes mellitus and its complications. Nature Reviews Endocrinology [Review Article]. 2017; 14:88. 3. Kameswaran V, Bramswig NC, McKenna LB, Penn M, Schug J, Hand NJ, et al. Epigenetic regulation of the DLK1-MEG3 microRNA cluster in human type 2 diabetic islets. Cell Metab 2014; 19(1):135-45. 4. Kwak SH, Park KS. Recent progress in genetic and epigenetic research on type 2 diabetes. Exp Mol Med 2016; 48:e220. 5. Nilsson E, Ling C. DNA methylation links genetics, fetal environment, and an unhealthy lifestyle to the development of type 2 diabetes. Clin Epigenetics 2017; 9:105. 6. Makki K, Froguel P, Wolowczuk I. Adipose tissue in obesity-related inflammation and insulin resistance: cells, cytokines, and chemokines. ISRN Inflamm 2013; 2013:139239 7. Kahn SE, Hull RL, Utzschneider KM. Mechanisms linking obesity to insulin resistance and type 2 diabetes. Nature 2006; 444(7121):840-6. 8. Nicholas DA, Andrieu G, Strissel KJ, Nikolajczyk BS, Denis GV. BET bromodomain proteins and epigenetic regulation of inflammation: implications for type 2 diabetes and breast cancer. Cell Mol Life Sci 2017; 74(2):231-43. 9. Sommese L, Zullo A, Mancini FP, Fabbricini R, Soricelli A, Napoli C. Clinical relevance of epigenetics in the onset and management of type 2 diabetes mellitus. Epigenetics 2017; 12(6):401-15. 10. Al Muftah WA, Al-Shafai M, Zaghlool SB, Visconti A, Tsai P-C, Kumar P, et al. Epigenetic associations of type 2 diabetes and BMI in an Arab population. Clinical epigenetics 2016; 8: 13. 11. Akash MSH, Rehman K, Liaqat A. Tumor Necrosis Factor-Alpha: Role in Development of Insulin Resistance and Pathogenesis of Type 2 Diabetes Mellitus. J Cell Biochem 2018; 119(1):105-10. 12. Miao F, Gonzalo IG, Lanting L, Natarajan R. In vivo chromatin remodeling events leading to inflammatory gene transcription under diabetic conditions. J Biol Chem 2004; 279(17):18091-7. 13. Hermsdorff HH, Mansego ML, Campión J, Milagro FI, Zulet MA, Martínez JA. TNF-alpha promoter methylation in peripheral white blood cells: Relationship with circulating TNFα, truncal fat and n-6 PUFA intake in young women. Cytokine 2013; 64(1):265-71. 14. Liu C, Feng X, Li Q, Wang Y, Li Q, Hua M. Adiponectin, TNF-alpha and inflammatory cytokines and risk of type 2 diabetes: A systematic review and meta-analysis. Cytokine 2016; 86:100-9. 15. Pinzon-Cortes JA, Perna-Chaux A, Rojas-Villamizar NS, Diaz-Basabe A, Polania-Villanueva DC, Jacome MF, et al. Effect of diabetes status and hyperglycemia on global DNA methylation and hydroxymethylation. Endocr Connect 2017; 6(8):708-25 مرکز تحقیقات سلولی و مولکولی، پژوهشکده‌ی علوم پایه‌ی سلامت Effat Farrokhi عفت فرخی e_farrokhi_k@yahoo.com Yes Department of Molecular Medicine, School of Advanced Technologies, Shahrekord University of Medical Sciences, Shahrekord, Iran گروه پزشکی مولکولی، دانشکده‌ی فناوری‌های نوین Morteza Hashemzadeh Chaleshtori مرتضی هاشم زاده چالشتری No Modeling in Health Center, Shahrekord University of Medical Sciences, Shahrekord, Iran مرکز تحقیقات مدل‌سازی در سلامت