بررسی اثرات مصرف پروتئین بالا بر بیان ژن TCF7L2 و آنزیم‌های آنتی‌اکسیدانی فیزیولوژیک در کبد مدل رت

نوع مقاله : مقاله پژوهشی

نویسندگان

1 دانشکده علوم، دانشگاه القاسم سبز، ۵۱۰۱۳، بابل، عراق.

2 دانشکده دامپزشکی، دانشگاه القاسم سبز، ۵۱۰۱۳، بابل، عراق.

3 دانشکده دامپزشکی، دانشگاه القاسم سبز، ۵۱۰۱۳، بابل، عراق

چکیده

هدف: تغذیه با پروتئین بالا با تغییرات عمیق در متابولیسم کبدی همراه است، با این حال سازوکارهای مولکولی ارتباط‌دهنده مصرف زیاد پروتئین با این تغییرات متابولیکی هنوز به‌طور کامل روشن نشده‌اند. هدف از این پژوهش، بررسی تأثیر رژیم غذایی پرپروتئین بر بیان ژن کبدی TCF7L2 و فعالیت آنزیم‌های آنتی‌اکسیدانی در رت‌های نژاد ویستار بود.
مواد و روش‌ها: در مجموع ۴۸ رت نر ویستار (سن ۸–۱۰ هفته، وزن اولیه ۲۰۰–۲۵۰ گرم) به‌صورت تصادفی به چهار گروه (هر گروه ۱۲ سر) تقسیم شدند: گروه شاهد (رژیم حاوی 10% پروتئین)، گروه کم‌پروتئین (5%)، گروه پروتئین متوسط (20%) و گروه پرپروتئین (40%). حیوانات به مدت ۱۲ هفته با رژیم‌های تعیین‌شده تغذیه شدند. فعالیت آنزیم‌های آنتی‌اکسیدانی کبدی شامل سوپراکسید دیسموتاز (SOD)، گلوتاتیون پراکسیداز (GPx) و نیتریک‌اکسید سنتاز اندوتلیال (eNOS) به روش اسپکتروفتومتری اندازه‌گیری شد. بیان ژن TCF7L2 با استفاده از PCR کمی در زمان واقعی (qRT-PCR) ارزیابی گردید. همچنین، میزان آلبومین و پروتئین تام سرم و وزن بدن حیوانات اندازه‌گیری شد. داده‌ها به‌صورت میانگین ± انحراف معیار گزارش شدند و تحلیل آماری با آزمون آنالیز واریانس یک‌طرفه (ANOVA) و آزمون تعقیبی توکی انجام گرفت.
نتایج: بیان mRNA ژن TCF7L2 در گروه شاهد به‌عنوان خط پایه (تغییر بیان = 1) در نظر گرفته شد. در گروه پرپروتئین، بیان این ژن نسبت به گروه شاهد به میزان 9/0±8/6 برابر افزایش یافت (p<0.001). یافته‌ها نشان داد که رژیم غذایی پرپروتئین به‌طور معنی‌داری موجب افزایش بیان ژن TCF7L2 شد (p<0.001). فعالیت آنزیم SOD در گروه پرپروتئین نسبت به شاهد 45% افزایش داشت (p<0.01). فعالیت GPx به میزان 38% افزایش یافت (p<0.01) و فعالیت eNOS نیز 52% افزایش نشان داد (p<0.001). همچنین، مقادیر پروتئین تام و آلبومین سرم در رت‌های دریافت‌کننده رژیم پرپروتئین به‌طور معنی‌داری بیشتر بود (p<0.05). کاهش معنی‌دار افزایش وزن بدن در گروه پرپروتئین نسبت به گروه‌های شاهد و کم‌پروتئین مشاهده شد (p<0.05).
نتیجه‌گیری: نتایج این پژوهش نشان می‌دهد که پروتئین غذایی می‌تواند از طریق تنظیم رونویسی وابسته به TCF7L2، سازوکارهای دفاع آنتی‌اکسیدانی کبد را تعدیل کند. رژیم پرپروتئین ممکن است از طریق فعالیت رونویسی ژن TCF7L2 در پیشگیری از بروز بیماری‌های متابولیک نقش داشته باشد. با این حال، انجام مطالعات بالینی بیشتر برای بررسی مزایای متابولیکی احتمالی این یافته‌ها در انسان ضروری است.

کلیدواژه‌ها

عنوان مقاله [English]

Study of the effects of high protein intake on TCF7L2 gene expression and physiological antioxidant enzymes in liver of rat model

نویسندگان [English]

  • Rawaa S. A. Al-Azawi 1
  • Hamzah H. K. Al-Shukri 2
  • Shaimaa Hussein Ali 1
  • Shatha M. Abbas 3
  • Nadya J. Ibraheem 3
  • Ekhlas Abid Hamza Al-Alwany 2
1 College of Science, Al-Qasim Green University, 51013, Babylon, Iraq.
2 College of Veterinary Medicine, Al-Qasim Green University,51013, Babylon, Iraq.
3 College of Veterinary Medicine, Al-Qasim Green University,51013, Babylon, Iraq.
چکیده [English]

Objective
High-protein feeding has been linked to profound changes in hepatic metabolism, although little has been shown regarding molecular mechanisms connecting high protein intake to these metabolic alterations. The present experiment examined the impact of high dietary protein on hepatic TCF7L2 gene expression and antioxidant enzyme activities in Wistar rats.
Materials and method
A total of 48 male Wistar rats (8-10 weeks old, initial body weight 200-250 g) were randomly assigned to four groups (12 per group): control (10% protein diet), low-protein (5% protein diet), moderate-protein (20% protein diet) and high-protein (40% protein diet) groups. Animals were then switched to their assigned diet and fed for 12 weeks. Hepatic levels of antioxidant enzyme activities (superoxide dismutase, SOD; glutathione peroxidase, GPx; endothelial nitric oxide synthase, eNOS) were determined spectrophotometrically. TCF7L2 gene expression was detected by quantitative real-time PCR. Levels of serum albumin and total protein were evaluated, as was body weight. Results are expressed as the mean ± S.D. Statistical analysis One-way ANOVA, followed by Tukey's multiple comparisons
Results
TCF7L2 mRNA expression in the control group was set as the baseline (fold-change = 1.0). The high-protein group demonstrated a 6.8±0.9-fold increase in TCF7L2 expression compared to control (p<0.001). The findings revealed that feeding a high protein diet significantly up-regulated the mRNA expression of TCF7L2 (p<0.001) compared to the control animals. The SOD activity was higher in the high protein group that CON, by 45% (p<0.01), the GPx activity increase by 38% (p<0.01) and eNOS activity showed a 52% increase (p<0.001). Serum total protein and albumin were markedly increased in rats fed high-protein diet (p<0.05). An important body weight gain reduction was evident in the high-protein group versus both control and low-protein groups (p<0.05).
Conclusion
These results imply that dietary protein might modulate hepatic antioxidant defense mechanisms mediated by TCF7L2-dependent transcriptions, and roles of high-protein in preventing the development of metabolic diseases mediating through transcriptional activity of TCF7L2. Further clinical studies are needed to investigate the potential metabolic benefits in humans as reported here from this rat model of disease.

کلیدواژه‌ها [English]

  • antioxidant enzymes
  • high protein diet
  • liver function
  • qRT-PCR
  • rat model

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مجله بیوتکنولوژی کشاورزی
دوره 18، شماره 2
خرداد 1405
صفحه 395-414

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