Effects of Food Restriction on PTGS2 and NRF2 Genes Expression Levels in Rat Testis

Abstract

This study aimed to investigate the effects of food restriction on oxidative status at the molecular levels of the testis in rats. A total of 16 male Wistar rats were allocated to two groups (n=8) and fed for four weeks. The first group was control (Cont) and consumed food and water ad libitum. The second one, food-restricted group (FR) was presented half of the regular consumption. But the water was presented ad libitum. The feeding period was maintained for four weeks. At the end of the feeding period, rats were euthanized by cardiac blood sampling under anesthesia. Malondialdehyde (MDA) levels in testis tissue were determined and calculated as nmol/mg protein. In addition, Prosta-glandin-Endoperoxide Synthase 2 (PTGS2) and Nuclear factor erythroid 2–related factor 2 (NRF2) genes expression levels were determined in testis tissue. While MDA and gene expression levels were found similar in groups, a positive correlation was found between PTGS2 and NRF2 genes (r=0.629; P<0.05). The molecular regulation of oxidative sta-tus was found strongly related with PTGS2 and NRF2 genes molecular activity in testis of rats. The obtained results were shown that the feeding period and restriction rate factors were mainly responsible for the oxidative status of testis tissue.

Keywords

• Food restriction
• NRF2
• oxidative stress
• PTGS2
• testis

Yem Kısıtlamasının Rat Testisinde PTGS2 ve NRF2 Genlerinin Ekspresyon Seviyelerine Etkileri

Abstract

Bu çalışmanın amacı, yem kısıtlamasının rat testisinde oksidatif duruma etkilerinin moleküler seviyede belirlenme-sidir. Toplam 16 erkek Wistar rat iki gruba ayrılmıştır (n=8) ve dört hafta boyunca beslenmiştir. Birinci grup olan Kontrol (Cont) grubuna yem ve su ad libitum olarak verilmiştir. İkinci grup olan, yem kısıtlaması (FR) grubuna Cont grubunun yarısı kadar yem sunulurken su ad libitum olarak sunulmuştur. Beslenme periyodunun sonunda ratlar anestezi altında kardiyak kan alınarak ötenazi edilmiştir. Testis dokusunda Malondialdehit (MDA) seviyeleri belirlenmiş ve nmol/mg protein olarak hesaplanmıştır. Ayrıca testis dokusunda Prostaglandin-Endoperoxide Synthase 2 (PTGS2) ve Nuclear factor erythroid 2–related factor 2 (NRF2) genlerinin ekspresyon seviyeleri tespit edilmiştir. Gruplarda MDA ve gen ekspresyon seviyeleri benzer bulunurken, PTGS2 ve NRF2 genleri arasında pozitif korelasyon bulunmuştur (r=0.629; P<0.05). Rat testisinde oksidatif durumun moleküler regülasyonu, PTGS2 ve NRF2 genlerinin moleküler aktivitesi ile güçlü bir şekilde ilişkili bulunmuştur. Elde edilen sonuçlar, testis dokusunun oksidatif durumundan esas olarak beslen-me süresi ve kısıtlama oranı faktörlerinin sorumlu olduğunu göstermiştir.

Keywords

• NRF2
• Oksidatif stres
• PTGS2
• testis
• yem kısıtlaması

References

1. Ajuogu PK, Al-Aqbi MA, Hart RA, Wolden M, Smart NA, McFarlane JR. The effect of dietary protein intake on factors associated with male infertility: A systematic literature review and meta-analysis of animal clinical trials in rats. Nutr Health 2020; 26(1): 53-64.
2. Alberts B, Johnson A, Lewis J, Roberts K, Walter P. Molecular Biology of the Cell. Fourth Edition. New York: Garland Science, 2002; p. 2568
3. Bozkaya F, Atli MO, Guzeloglu A, Kayis SA, Yildirim ME, Kurar E, Yilmaz R, Aydilek N. Effects of long‐term heat stress and dietary restriction on the ex-pression of genes of steroidogenic pathway and small heat‐shock proteins in rat testicular tissue. Andrologia 2017; 49(6): e12668.
4. Bruss MD, Khambatta CF, Ruby MA, Aggarwal I, Hellerstein MK. Calorie restriction increases fatty acid synthesis and whole body fat oxidation rates. Am J Physiol Metab 2010; 298(1): 108-16.
5. Filaire E, Rouveix M, Massart A, Gladine C, Davicco MJ, Durand D. Lipid peroxidation and antioxidant status in rat: effect of food restriction and wheel running. Eur J Appl Physiol 2009; 107(2): 243-50.
6. Gardner CD, Kim S, Bersamin A, Dopler-Nelson M, Otten J, Oelrich B, Cherin R. Micronutrient quality of weight-loss diets that focus on macronutrients: results from the A TO Z study. Am J Clin Nutr 2010; 92(2): 304-12.
7. Güvenç M, Cellat M, Özkan H, Tekeli İO, Uyar A, Gökçek İ, İşler CT, Yakan A. Protective effects of tyrosol against DSS-induced ulcerative colitis in rats. Inflammation 2019; 42(5): 1680-91.
8. Hamden K, Silandre D, Delalande C, ElFeki A, Car-reau S. Protective effects of estrogens and caloric restriction during aging on various rat testis param-eters. Asian J Androl 2008; 10(6): 837-45.

9. Harper M, Bevilacqua L, Hagopian K, Weindruch R, Ramsey JJ. Ageing, oxidative stress, and mito-chondrial uncoupling. Acta Physiol Scand 2004; 182(4): 321-31.
10. Ichikawa M, Fujita Y, Ebisawa H, Ozeki T. Effects of long-term, light exercise under restricted feeding on age-related changes in physiological and metabolic variables in male Wistar rats. Mech Ageing Dev 2000; 113(1); 23-35.
11. Jayroe J, Soulsby M, Chowdhury P. Attenuation of tissue oxidative stress by dietary restriction in rats on simulated microgravity. Ann Clin Lab Sci 2012; 42(2): 140-4.
12. Kim JD, McCarter RJM, Yu BP. Influence of age, exercise, and dietary restriction on oxidative stress in rats. Aging Clin Exp Res 1996; 8(2): 123-9.
13. Li Yinlam, Zhang L, Zheng X, Qian J, Li Yanquan, Xie C, Zhang X, Zhou Y, Huang H. Dietary restriction and/or exercise training impairs spermatogenesis in normal rats. Appl Physiol Nutr Metab 2021; 46(3): 229-37.
14. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the Folin phenol rea-gent. J Biol Chem 1951; 193(1): 265-75.
15. Mladenovic Djordjevic A, Loncarevic-Vasiljkovic N, Gonos ES. Dietary restriction and oxidative stress: Friends or enemies? Antioxid Redox Signal 2021; 34(5): 421-38.
16. Mortazavi M, Salehi I, Alizadeh Z, Vahabian M, Roushandeh AM. Protective effects of antioxidants on sperm parameters and seminiferous tubules epithelium in high fat-fed rats. J Reprod Infertil 2014; 15(1): 22.
17. Moszak M, Szulińska M, Bogdański P. You are what you eat-the relationship between diet, microbiota, and metabolic disorders-A review. Nutrients 2020; 12(4): 1096.
18. NRC. Nutrient Requirements of Laboratory Animals. Fourth Revised Edition. Washington (DC): National Academies Press (US) 1995; 11-58.
19. Onodera Y, Teramura T, Takehara T, Shigi K, Fuku-da K. Reactive oxygen species induce PTGS2 ex-pression via TAK1 activation in synovial fibroblast cells. FEBS Open Bio 2015; 5: 492-501.
20. Özkan H, Kutlu T. The relationship of fructose con-sumption with MDA levels in rat liver and its effect on the expression levels of PTGS2 and NRF-2 genes. Ankara Üniv Vet Fak Derg 2020; 67(4): 387-92.
21. Özkan H, Yakan A. Dietary high calories from sun-flower oil, sucrose and fructose sources alters lipo-genic genes expression levels in liver and skeletal muscle in rats. Ann Hepatol 2019; 18(5): 715-24.
22. Ross D, Siegel D. Functions of NQO1 in cellular pro-tection and CoQ10 metabolism and its potential role as a redox sensitive molecular switch. Front Physiol 2017; 8: 1-10.
23. Smyers ME, Bachir KZ, Britton SL, Koch LG, Novak CM. Physically active rats lose more weight during calorie restriction. Physiol Behav 2015; 139: 303-13.
24. Wang X, Shen CL, Dyson MT, Eimerl S, Orly J, Hut-son JC, Stocco DM. Cyclooxygenase-2 regulation of the age-related decline in testosterone biosyn-thesis. Endocrinology 2005; 146(10): 4202-8.
25. Wasowicz W, Neve J, Peretz A. Optimized steps in fluorometric determination of thiobarbituric acid-reactive substances in serum: importance of ex-traction pH and influence of sample preservation and storage. Clin Chem 1993; 39(12): 2522-6.
26. Yang J, Hou X, Gao A, Wang H. Effect of dietary energy and protein restriction followed by realimen-tation on pituitary mRNA expression of growth hor-mone and related genes in lambs. Small Rumin Res 2014; 119(1-3): 39-44.
27. Yang SH, Yu LH, Li L, Guo Y, Zhang Y, Long M, Li P, He JB. Protective mechanism of sulforaphane on cadmium-induced sertoli cell injury in mice testis via Nrf2/ARE signaling pathway. Molecules 2018; 23(7): 1774.