نقش رازیانه بر بیان ژن DLK1 در بافت قلب گوسفند

نوع مقاله : مقاله پژوهشی

نویسندگان

1 بخش علوم دامی، دانشکده کشاورزی، دانشگاه شهید باهنر کرمان

2 دانشجوی کارشناسی ارشد بخش علوم دامی، دانشکده کشاورزی، دانشگاه شهید باهنر کرمان، ایران.

3 بخش علوم دامی، دانشگاه شهید باهنر کرمان

4 استادیار، بخش زیست شناسی، دانشکده علوم، دانشگاه شهید باهنر کرمان، ایران

5 آزمایشگاه کلیدی اصلاح و تولید مثل ژنتیکی حیوانات، وزارت کشاورزی و امور روستایی، موسسه علوم دامی، آکادمی علوم کشاورزی چین، پکن 100193، چین

چکیده

هدف: رازیانه در رژیم‌های غذایی حیوانات برای اهداف مختلفی از جمله بهبود افزایش وزن، کیفیت اکسیداتیو گوشت، هضم و بازده رشد، حجم سلول‌های بسته، طول و وزن روده کوچک، کارایی لاشه، مصرف خوراک، گلبول‌های قرمز و تعداد هموگلوبین، تبدیل خوراک، کارایی و وضعیت سلامت و کاهش تعداد کل باکتری‌ها استفاده شده است. از طرفی ژن DLK1 نقش مه‌می‌در کنترل فرآیندهای مختلف در سرتاسر دوران جنینی و بزرگسالی دارد. لذا، هدف این مطالعه بررسی اثر رازیانه بر بیان ژن DLK1 در قلب گوسفند کرمانی با استفاده از real time PCR بود.
مواد و روش‌ها: در این پژوهش30 راس بره نر 6 ماهه از نژاد کرمانی با وزن تقریباً یکسان انتخاب و به طور تصادفی در سه گروه آزمایشی (10 راس در هر گروه) قرار گرفتند. حیوانات آزمایشی به مدت سه ماه  با سه سطح رازیانه (0، 1 و 2 درصد) به صورت آزاد و جداگانه تغذیه شدند. پس از کشتار وزن قلب و چربی اطراف قلب اندازه گیری و نمونه برداری از بافت‌ قلب انجام شد. استخراج RNA و ساخت cDNA با استفاده از کیت‌های استاندارد صورت گرفت. با استفاده از آغازگرهای مربوطه بیان ژن‌های DLK1 (ژن هدف) و  ژن بتا اکتین (ژن مرجع) با استفاده از روش real-time PCR انجام و نتایج با نرم‌افزارهای مربوطه آنالیز شد.
نتایج: نتایج بررسی کیفیت RNA­های استخراج شده روی ژل آگارز و در طول موج A260/A280 نشان دهنده کیفیت مناسب و مطلوب RNA کل بود. نتایج  real time PCR  و الکتروفورز محصولات PCR روی ژل آگارز نشان داد که ژن DLK1  در بافت قلب بیان شده است. افزودن رازیانه به جیره باعث تغییر معنی‌دار وزن قلب و چربی اطراف قلب نسبت به جیره شاهد نشد. نتایج نشان داد که تغذیه رازیانه در سطوح 1% و 2% به طور معنی داری (P<0.05) بیان ژن DLK1 را در بافت قلب افزایش می‌دهد.
نتیجه‌گیری: بر اساس نتایج می‌توان نتیجه گرفت که رازیانه ‌می‌تواند در رژیم غذایی گوسفند برای بهبود ساختار قلب از طریق اثرات مثبت بر بیان ژن DLK1 استفاده شود. از آنجایی که رازیانه سطح بیان ژن DLK1 را در بافت قلب افزایش داده است، می‌توان آن را برای بهبود عملکرد قلب در نظر گرفت. می‌توان نتیجه گرفت که رازیانه می‌تواند برای مصارف مختلفی در دام استفاده شود، اما برای هر اثر در هر بافت، تحقیقات بیشتری با در نظر گرفتن شرایط مختلف ژنتیکی، اپی‌ژنتیکی و فیزیولوژیکی انجام شود تا به نتیجه نهایی برسیم.

کلیدواژه‌ها


عنوان مقاله [English]

The role of fennel on DLK1 gene expression in sheep heart tissue

نویسندگان [English]

  • Mohammadreza Mohammadabadi 1
  • Diana Shaban Jorjandy 2
  • Zahra Arabpoor Raghabadi 3
  • Fatemeh Abareghi 2
  • Hosein Ali Sasan 4
  • Farhad Bordbar 5
1 Department of Animal Science, Faculty of Agriculture, College of Agriculture, Shahid Bahonar University of Kerman
2 MSc Student, Animal Science Department, Faculty of Agriculture, Shahid Bahonar University of Kerman, Kerman, Iran.
3 MSc Student, Animal Science Department, Faculty of Agriculture, Shahid Bahonar University of Kerman, Kerman, Iran
4 Assistant professor, Department of Biology, Faculty of Science, Shahid Bahonar University of Kerman, Kerman, Iran.
5 Key Laboratory of Animal Genetics Breeding and Reproduction, Ministry of Agriculture and Rural Affairs, Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, Beijing 100193, China
چکیده [English]

Objective
Fennel in animal diets for various purposes including improving weight gain, oxidative quality of meat, digestion and growth efficiency, closed cell volume, small intestine length and weight, carcass efficiency, feed intake, red blood cells and hemoglobin count, feed conversion, efficiency and health status and reduction of the total number of bacteria have been used. On the other hand, DLK1 gene plays an important role in controlling various processes throughout the fetus and adulthood. Therefore, the aim of this study was to investigate the effect of fennel on DLK1 gene expression in Kermani sheep heart using real-time PCR.
Materials and Methods
In this study, 30 male Kermani lambs (6-month-old) with almost the same weight were selected and randomly divided into three experimental groups (10 in each group). Experimental animals were fed freely and separately with three levels of fennel (0, 1 and 2%) for three months. After slaughtering, the heart weight and fat around the heart were measured and the heart tissue was sampled. RNA extraction and cDNA synthesis were performed using standard kits. The expression of DLK1 genes (target gene) and beta-actin gene (reference gene) was performed using proper primers using real-time PCR method and the results were analyzed with the appropriate software.
Results
The results of quality assay of extracted RNAs on agarose gel and at A260 / A280 wavelength showed good quality of total RNA. The results of real-time PCR and electrophoresis of PCR products on agarose gel showed that DLK1 gene was expressed in heart tissue. Adding fennel to the diet did not significantly change the weight of the heart and the weight of fat around the heart compared to the control diet. The results showed that fennel nutrition at 1% and 2% levels significantly (P <0.05) increased DLK1 gene expression in heart tissue.
Conclusions
Based on the results, it can be concluded that fennel can be used in sheep diet to improve heart structure through positive effects on DLK1 gene expression. Since fennel has increased the expression level of DLK1 gene in heart tissue, it can be considered to improve heart function. It can be concluded that fennel can be used for different purposes in livestock, but for each effect in each tissue, more research should be done considering different genetic, epigenetic and physiological conditions to reach the distinct conclusion.

کلیدواژه‌ها [English]

  • DLK1 gene expression
  • fennel
  • diet
  • heart
  • heart muscle
محمدآبادی محمدرضا (1399) بیان ژن ESR1 در بز کرکی راینی با استفاده از real time PCR‎. مجله بیوتکنولوژی کشاورزی 12(1)، 192-177.
محمدآبادی محمدرضا (1399) پروفایل بیانی mRNA مختص بافت ژن ESR2  در بز‎. مجله بیوتکنولوژی کشاورزی 12(2)، 184-169.
محمدآبادی محمدرضا، کرد محبوبه، نظری محمود (1397) مطالعه بیان ژن لپتین در بافت‌های مختلف گوسفند کرمانی با استفاده از real time PCR. مجله بیوتکنولوژی کشاورزی 10(3)، 122-111. 
محمدی فر آمنه، محمدآبادی محمدرضا (1390) کاربرد نشانگرهای ریزماهواره برای مطالعه ژنوم گوسفند کرمانی. مجله علوم دا‌می‌ایران 42(4)، 344-337.
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