بهینه نمودن رنگ‌آمیزی اکریدین اورنج جهت ارزیابی تأثیر حفاظتی زعفران و ویتامین E بر ساختار DNA هسته‌ی اسپرم رت

نوع مقاله : Original Article(s)

نویسندگان

1 استاد، گروه علوم تشریح و بیولوژی مولکولی، دانشکده‌ی پزشکی، دانشگاه علوم پزشکی اصفهان، اصفهان، ایران

2 دانشجوی دکتری، گروه مهندسی پزشکی، دانشکده‌ی پزشکی، دانشگاه علوم پزشکی تهران، تهران، ایران

3 دانشیار، گروه علوم تشریح و بیولوژی مولکولی، دانشکده‌ی پزشکی، دانشگاه علوم پزشکی اصفهان، اصفهان، ایران

چکیده

مقدمه: در دهه‌های اخیر، ارتباط بین غلظت ROS (Reactive oxygen species) و کیفیت مایع Semen مورد توجه قرار گرفته و تأثیر آن بر ناباروری در مطالعات متعددی بررسی شده است. زعفران نه تنها به عنوان یک افزودنی غذایی، بلکه به طور سنتی به عنوان یک گیاه دارویی که دارای خاصیت آنتی‌اکسیدانی است، همواره مورد توجه بوده است. ویتامین E نیز آنتی‌اکسیدان بسیار قوی است و قادر است سلول و اجزای آن را در مقابل هجوم آنتی‌اکسیدان‌ها محافطت نماید. هدف از این مطالعه، بررسی اثر آنتی‌اکسیدانی زعفران و ویتامین E در حفاظت از دناتوراسیون ساختار کروماتین اسپرم در مواجهه با اسید بود.روش‌ها: تعداد 30 رت نر بالغ ویستار به صورت اتفاقی و مساوی به سه گروه شاهد (cc/day 5/0آب مقطر روزانه)، زعفران (mg/kg/day 100) و ویتامین E (mg/kg/day 100) تقسیم شدند. پس از 60 روز دوره‌ی تیمار، اپیدیدیم جدا شد و از اسپرم‌های ناحیه‌ی دم آن، جهت بررسی مقاومت ساختار DNA در مقابل اسید دترجنت با استفاده از رنگ‌آمیزی آکریدین اورنج (OA یا Acridine orange) استفاده شد.یافته‌ها: زعفران و ویتامین E باعث کاهش حساسیت DNA به آسیب توسط اسید دترجنت می‌شوند (001/0 > P).نتیجه‌گیری: احتمال می‌رود زعفران و ویتامین E به دلیل پتانسیل آنتی‌اکسیدانی بالای خود، قادر باشند ساختار کروماتین را از اثرات تخریبی اسید دترجنت محافظت نمایند. 

کلیدواژه‌ها


عنوان مقاله [English]

Optimizing Acridine Orange Staining for Assessment of Protective Effects of Saffron and Vitamin E on Rat Sperm DNA Structure

نویسندگان [English]

  • Shahnaz Razavi 1
  • Sayyed Ahmad Vaez 2
  • Mohammad Mardani 3
1 Professor, Department of Anatomical Sciences and Molecular Biology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran
2 PhD student, Department of Tissue Engineering, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran
3 Associate Professor, Department of Anatomical Sciences and Molecular Biology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran
چکیده [English]

Background: In recent decades, relation between reactive oxygen species (ROS) concentration and semen quality was noted. Traditionally, saffron has been not only considered as a food additive but also as a medicinal herb, which has good antioxidant properties. Vitamin E is considered as a endogenous and supplementary antioxidant which can protects the cell. The aim of this study was to evaluate the protection influence of saffron and vitamin E on sperm DNA against acid.Methods: Thirty adult male Wistar rats randomly divided into 3 equal groups of saffron, vitamin E and controlwhich received saffron (100 mg/kg/day), vitamin E (100 mg/kg/day) and distilled water (0.5 cc/day), respectively. After 60 days, cauda epididymis dissected and sperm were used for analysis of sperm chromatin susceptibility to acid denaturation via acridine orange (AO) staining. Acridine orange staining was carried out with or without acid detergent incubation.Findings: Both, saffron and vitamin E, decreased sperm DNA damage against acid (P < 0.001).Conclusion: Saffron and vitamin E protect DNA against denaturation, probably because of their powerful antioxidant properties.

کلیدواژه‌ها [English]

  • Antioxidants
  • Saffron
  • Vitamin E
  • DNA damage
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