بررسی الگوی الکتروفورتیک پروتئینها و محافظت از استرس اکسیداتیو مایع فولیکولی زنان سالم و نازا

نوع مقاله: اصیل پژوهشی

نویسندگان

1 دانشجوی کارشناسی ارشد بیوشیمی بالینی، دانشگاه آزاد اسلامی، واحد شاهرود، شاهرود، ایران.

2 استاد گروه بیوشیمی، دانشکده پزشکی، دانشگاه آزاد اسلامی، واحد شاهرود، شاهرود، ایران.

3 استادیار گروه زنان و مامایی، دانشکده پزشکی، دانشگاه آزاد اسلامی، واحد شاهرود، شاهرود، ایران.

10.22038/ijogi.2019.14004

چکیده

مقدمه: مطالعات اپیدمیولوژی نشان می­دهد که 15-10% از زوجین در دنیا ناباروری را تجربه می­کنند. برآورد می­شود که تقریباً 50-40% از مشکلات نازایی منشأ زنانه و 30% موارد منشأ مردانه دارند و بقیه مربوط به هر دو نفر می­باشد. مایع فولیکولی، محیطی را برای توسعه تخمک‌گذاری ایجاد می­کند و منعکس‌کننده فعالیت­های بیوشیمیایی و هورمونی فولیکول­ها در اندازه­های متفاوت می­باشد. تأثیر استرس اکسیداتیو مایع فولیکول بر بلوغ تخمک، بارور شدن و بارداری به‌طور کامل مشخص نیست، لذا مطالعه حاضر با هدف بررسی و مقایسه خاصیت آنتی‌اکسیدانی مایع فولیکولی در زنان سالم و نازا و همچنین بررسی الگوی الکتروفورزی پروتئین­های مایع فولیکی در این دو گروه انجام شد.
روش‌کار: این مطالعه مورد- شاهدی و مقطعی در سال 1397 بر روی 17 زن نازا با اختلال تخمک‌گذاری و 17 زن سالم با نازایی به دلیل اختلال مردانه کاندید IVF مراجعه کننده به مرکز ناباروری فاطمه الزهراء بابل انجام شد. نمونه­های مایع فولیکولی از 34 زن نازا و سالم استخراج گردید. خاصیت آنتی‌اکسیدانی با روش‌های FRAP (بررسی توان آنتی اکسیدانی به روش احیای فریک) و DPPH (دی فنیل پیکریل هیدرازیل) سنجش شد و میزان پراکسیداسیون لیپیدی با سنجش مالون دی آلدئید (MDA) تعیین شد. جهت بررسی الگوی الکتروفورز پروتئین­ها از الکتروفورز عمودی SDS PAGE (الکتروفورز روی ژل پلی آکریل آمید در حضور سدیم دو- دسیل سولفات) و جهت سنجش پروتئین توتال از روش برادفورد استفاده شد. تجزیه و تحلیل داده­ها با استفاده از نرم‌افزار آماری SPSS (نسخه 16) و آزمون تی انجام شد. میزان p کمتر از 05/0 معنی‌دار در نظر گرفته شد.
یافته‌ها: بر اساس نتایج، خاصیت آنتی‌اکسیدانی (23/0=p) و پراکسیداسیون لیپیدی (45/0=p) در دو گروه اختلاف معناداری را نشان نداد. همچنین نتایج تست الکتروفورز و پروتئین توتال نیز در دو گروه اختلاف معنی‌داری نداشت (73/0=p). بیشترین میزان پروتئین­ها در هر دو گروه در باند 75-48 کیلو دالتون مشاهد شد.
نتیجه‌گیری: اگرچه بین دو گروه مورد مطالعه در شاخص­های استرس اکسیداتیو اختلاف وجود داشت، ولی این اختلاف از نظر آماری معنی‌دار نبود که دلیل آن را می­توان به کم بودن تعداد نمونه در هر گروه نسبت داد. جهت نشان دادن اختلاف در الگوی الکتروفورزی پروتئین­های مایع فولیکولی نیاز به بررسی دقیق‌تر با استفاده از الکتروفورز دوبٌعدی وجود دارد.

کلیدواژه‌ها


عنوان مقاله [English]

Protein Electrophoresis Pattern and Oxidative Stress Protection of Human Follicular Fluid in Fertile and Infertile Women

نویسندگان [English]

  • Maryam Hasanpour 1
  • Mohammad Taghi Goodarzi 2
  • Mahnaz Nouri 3
1 M.Sc. Student, Department of Clinical Biochemistry, Shahrood Branch, Islamic Azad University, Shahrood, Iran
2 Professor, Department of Biochemistry, Faculty of Medicine, Shahrood Branch, Islamic Azad University, Shahrood, Iran
3 Assistant Professor, Department of Obstetrics and Gynecology, Faculty of Medicine, Shahrood Branch, Islamic Azad University, Shahrood, Iran.
چکیده [English]

Introduction: Epidemiological studies show that 10 to 15% of couples experience infertility worldwide. It is estimated that 40-50% of infertilities are related to female factors and 30% are male factors and others are attributed to both partners. Follicular fluid supplies an environment for ovulation and it reflects hormonal and biochemical activities of follicles in different stages. The effects of follicular fluid oxidative stress on ovum maturation and fertility are not well explained. Therefore, this study was performed with aim to evaluate and compare antioxidant properties of follicular fluid in healthy and infertile women and also, to examine the protein electrophoresis pattern of follicular fluid in these two groups. 
Methods: This case-control and cross-sectional study was performed on 17 infertile women with ovulation disorder and 17 healthy women with infertility related to male factor who were candidate for IVF and had referred to Fatemeh-Zahra infertility center, Babol in 2018. Follicular fluid specimens were collected from 34 infertile and healthy women. To measure antioxidant properties, FRAP (Ferric reducing antioxidant power) and DPPH (1,1-Diphenyl-2-picryl-hydrazyl) tests were performed. Lipid peroxidation was assayed using MDA (Malondialdehyde) test. SDS-PAGE (sodium dodecyl sulphate-polyacrylamide gel electrophoresis) was used to evaluate protein electrophoresis pattern. Bradford method was used to assess total protein. Data were analyzed by SPSS software (version 16) and t-test. P<0.05 was considered statistically significant.
Results: Antioxidant properties and lipid peroxidation assay did not show any statistical significant differences between two groups (p=0.23 and p=0.45, respectively). Also, the results of electrophoresis test and total protein were not significantly different between two groups (p=0.73). Most amount of proteins in both groups was observed in heavy band between 48 -75 Kilo Dalton region.
Conclusion: Although there was difference in oxidative stress level between two groups, it was not statistically significant. It can be attributed to small sample size in each group. More close evaluation using 2-dimentional electrophoresis is needed to show difference in follicular fluid protein electrophoresis pattern.

کلیدواژه‌ها [English]

  • female infertility
  • follicular fluid
  • Oxidative stress
  • Protein electrophoresis
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