Aralıklı Oruç Koşullarında Kolon Dokusunda Apoptozisin Gen Düzeyinde Modülasyonu

Öz

Aralıklı açlık (IF), apoptoz dâhil olmak üzere hücresel stres yolları üzerinde belirgin etkiler gösteren bir metabolik strateji olarak giderek daha fazla kabul görmektedir. Bu çalışma, aralıklı açlık protokolü sonrasında kolon dokusunda temel apoptoz genlerinin transkripsiyonel düzenlenmesini incelemiştir. On iki aylık erkek Wistar sıçanları, 35 gün boyunca günlük 18 saatlik açlık rejimine tabi tutulmuştur. Kontrol grubu yem ve suya ad libitum erişime sahipti. Anti-apoptotik genler olan B-cell lymphoma 2 (BCL-2) ve B-cell lymphoma-extra-large (BCL-XL) ile pro-apoptotik düzenleyiciler olan BCL-2 homologous antagonist/killer (BAK), BCL-2-associated X protein (BAX) ve Cysteine-aspartic acid protease 3 (Caspase-3) genlerinin mRNA ekspresyon düzeyleri, kantitatif gerçek zamanlı PCR ile analiz edilmiştir. Aralıklı açlık, BCL-2 ve BCL-XL genlerinin ekspresyonunu anlamlı düzeyde artırırken, BAK, BAX ve Caspase-3 ekspresyonlarını ise belirgin şekilde azaltmıştır. Bu bulgular, aralıklı açlığın kolon dokusunda apoptotik duyarlılığı azaltan bir transkripsiyonel değişimi tetiklediğini ve böylece hücresel koruyucu bir fenotip oluşturduğunu göstermektedir. Sonuçlar, aralıklı açlığın yaşlanma sürecinde kolon dokusunun dayanıklılığını artırmaya yönelik farmakolojik olmayan bir müdahale olarak potansiyelini vurgulamakta ve moleküler düzeyde yaşa bağlı apoptoz ve metabolik düzensizliklerin hafifletilmesinde uygulanabilirliğini desteklemektedir.

Anahtar Kelimeler

• Aralıklı açlık
• Apoptoz
• Bcl-2 ailesi genleri
• Kolon gen ekspresyonu
• Wistar sıçanı

Gene-Level Modulation of Apoptosis in the Colon under Intermittent Fasting Conditions

Öz

Intermittent fasting (IF) has gained recognition as a metabolic strategy with significant impacts on cellular stress pathways, including apoptosis. This study examined the transcriptional regulation of key apoptotic genes in colon tissue following an intermittent fasting regimen. Twelve-month-old male Wistar rats were subjected to an 18-hour daily fasting protocol for 35 consecutive days. The control group had ad libitum access to food and water. Quantitative real-time PCR analysis was employed to evaluate mRNA expression levels of anti-apoptotic genes B-cell lymphoma 2 (BCL-2) and B-cell lymphoma-extra-large (BCL-XL) and pro-apoptotic mediators BCL-2 homologous antagonist/killer (BAK), BCL-2-associated X protein (BAX), and Cysteine-aspartic acid protease 3 (Caspase-3). IF significantly increased BCL-2 and BCL-XL expression while concurrently downregulating BAK, BAX, and Caspase-3. These findings indicate that intermittent fasting induces a transcriptional shift that reduces apoptotic susceptibility in colon, thereby promoting a cytoprotective phenotype. The results underscore IF as a potential non-pharmacological intervention to enhance colonic resilience during aging, supporting its application in mitigating age-related apoptotic and metabolic dysregulation at the molecular level.

Anahtar Kelimeler

• Intermittent fasting
• Apoptosis
• Bcl-2 family genes
• Colon gene expression
• Wistar rat

Kaynakça

1. Shetty, A. K., Kodali, M., Upadhya, R., et al. (2018). Emerging anti-aging strategies - scientific basis and efficacy. Aging and Disease, 9, 1165-1184.
2. Mattson, M. P., Longo, V. D., & Harvie, M. (2017). Impact of intermittent fasting on health and disease processes. Ageing Research Reviews, 39, 46-58.
3. de Cabo, R., & Mattson, M. P. (2019). Effects of intermittent fasting on health, aging, and disease. New England Journal of Medicine, 381, 2541-2551.
4. Hovik, F. S., & Longo, V. D. (2024). Fasting-mimicking diets in longevity and disease. In K. Varady, E. N. C. Manoogian, & V. D. Longo (Eds.), Intermittent and periodic fasting, aging and disease (pp. 225-242). Cham: Springer Nature Switzerland.
5. Rangan, P., Choi, I., Wei, M., et al. (2019). Fasting-mimicking diet modulates microbiota and promotes intestinal regeneration to reduce inflammatory bowel disease pathology. Cell Reports, 26, 2704-2719.e6.
6. Mohammadzadeh, A., Roshanravan, N., Mesri Alamdari, N., et al. (2021). The interplay between fasting, gut microbiota, and lipid profile. International Journal of Clinical Practice, 75, e14591.
7. Teker, H. T., & Ceylani, T. (2023). Intermittent fasting supports the balance of the gut microbiota composition. International Microbiology, 26, 51-57.
8. Ardahanlı, İ., Özkan, H. İ., Özel, F., et al. (2022). Infrared spectrochemical findings on intermittent fasting-associated gross molecular modifications in rat myocardium. Biophysical Chemistry, 289, 106873.

9. Ceylani, T., Teker, H. T., Samgane, G., et al. (2022). Intermittent fasting-induced biomolecular modifications in rat tissues detected by ATR-FTIR spectroscopy and machine learning algorithms. Analytical Biochemistry, 654, 114825.
10. Elmore, S. (2007). Apoptosis: A review of programmed cell death. Toxicologic Pathology, 35, 495-516.
11. Cory, S., & Adams, J. M. (2002). The Bcl2 family: Regulators of the cellular life-or-death switch. Nature Reviews Cancer, 2, 647-656.
12. Rifaai, R. A., El-Tahawy, N. F. G., Abozaid, S. M. M., et al. (2025). Intermittent fasting ameliorates age-induced morphological changes in aged albino rat kidney via autophagy activation and reduction of apoptosis and inflammation. Microscopy and Microanalysis, 31, 1-12.
13. Mendonça, M. L. M., Carvalho, M. R., Romanenghi, R. B., et al. (2024). Impact of combined intermittent fasting and high-intensity interval training on apoptosis and atrophy signaling in rat fast- and slow-twitch muscles. Physiological Reports, 12, 1-15.
14. Hafez, S. M. N. A., & Elbassuoni, E. (2022). Dysfunction of aged liver of male albino rats and the effect of intermitted fasting: Biochemical, histological, and immunohistochemical study. International Immunopharmacology, 103, 108465.
15. Strilbytska, O., Klishch, S., Storey, K. B., et al. (2024). Intermittent fasting and longevity: From animal models to implication for humans. Ageing Research Reviews, 96, 102274.
16. Ceylani, T., Taner, H., Samgane, G., et al. (2022). Intermittent fasting-induced biomolecular modifications in rat tissues detected by ATR-FTIR spectroscopy and machine learning algorithms. Analytical Biochemistry, 654, 114825.
17. Livak, K. J., & Schmittgen, T. D. (2001). Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods, 25, 402-408.
18. Schmittgen, T. D., & Livak, K. J. (2008). Analyzing real-time PCR data by the comparative CT method. Nature Protocols, 3, 1101-1108.
19. Aba, N., Koçpınar, E. F., & Ceylani, T. (2023). The hepatic antioxidant system damage induced with the cafeteria (CAF) diet is largely counteracted using SCD probiotics during development of male Wistar rats. Nutrients, 15, 4557.
20. Baba, B., Ceylani, T., Gurbanov, R., et al. (2024). Promoting longevity in aged liver through NLRP3 inflammasome inhibition using tauroursodeoxycholic acid (TUDCA) and SCD probiotics. Archives of Gerontology and Geriatrics, 125, 105517.
21. Guo, R., & Li, G. (2018). Tanshinone modulates the expression of Bcl-2 and Bax in cardiomyocytes and has a protective effect in a rat model of myocardial ischemia-reperfusion. Hellenic Journal of Cardiology, 59, 323-328.
22. Valks, D. M., Kemp, T. J., & Clerk, A. (2003). Regulation of Bcl-xL expression by H2O2 in cardiac myocytes. Journal of Biological Chemistry, 278, 25542-25547.
23. Zhang, J., He, J., Xia, J., et al. (2012). Delayed apoptosis by neutrophils from COPD patients is associated with altered Bak, Bcl-xl, and Mcl-1 mRNA expression. Diagnostic Pathology, 7, 65.
24. Lin, H. H., Lin, S. M., Chung, Y., et al. (2014). Dynamic involvement of ATG5 in cellular stress responses. Cell Death & Disease, 5, e1478.
25. Kwon, N., Lee, K. E., Singh, M., et al. (2021). Suitable primers for GAPDH reference gene amplification in quantitative RT-PCR analysis of human gene expression. Gene Reports, 24, 101272.