Changes in Histological Features, Apoptosis and Necroptosis, and Inflammatory Status in the Livers and Kidneys of Young and Adult Rats

Year 2024, , 85 - 96, 30.05.2024

Emine Rümeysa Hekimoğlu , Mukaddes Eşrefoğlu , Birsen Elibol , Seda Kırmızıkan

https://doi.org/10.26650/EurJBiol.2024.1471005

Abstract

Objective: Aging entails a gradual rise in low-grade inflammation affected by cellular degeneration and death. Inflammaging refers to the chronic, low-grade inflammation that occurs alongside the aging process. This study attempts to evaluate the hepatic and renal histological changes, apoptosis and necroptosis rates, and inflammaging status of 6-week-old and 10-month-old rats.
Materials and Methods: This study uses 12 male rats separated into two groups: Young Group (6-week-old rats; n = 6), and Adult Group (10-month-old rats; n = 6). Animals were sacrificed under anesthesia. The rats’ livers and kidneys were removed, and each organ tissue was divided into two parts: one for the microscopic examination (H&E and TUNEL immunohistochemistry) and the other for biochemical determination (tumor necrosis factor-alpha [TNF-α], nuclear factor-kappa beta [NF-𝜅B], interleukin1-Beta [IL-1β], IL6, receptor-interacting serine/threonine protein kinase [RIP], and RIP3).
Results: The histological features of the livers and kidneys of the 6-week-old rats were consistent with healthy mammalian organ features, while some histological changes were detected in sections of the 10-month-old rats. The apoptosis rate indicated by TUNEL immunohistochemistry was seen to have increased in the 10-month-old rats, while the necroptosis rate indicated by RIP3 Western-blotting analysis was conversely determined to have decreased. Significant increases in TNF-α and NF-𝜅B levels were consistent with the increased apoptosis rate in the 10-month-old rats compared to the 6-week-old rats.
Conclusion: One of the striking results of this study is that the degenerative changes related to aging began to be seen even in 10-month-old rats. The researchers used healthy rats of this age as control subjects as well as to create experimental models.

Keywords

Apoptosis, Necroptosis, Young rats, Adult rats, Liver, Kidney

Ethical Statement

The Animal Care and Experiment Committee of Bezmialem Vakif University, School of Medicine approved the animal care and research procedure (No: 2024/10).

References

• Acosta JC, Gil J. Senescence: A new weapon for cancer therapy. Trends Cell Biol. 2012;22(4):211-219. google scholar
• Coppe JP, Desprez PY, Krtolica A, Campisi J. The senescence-associated secretory phenotype: The dark side of tumor suppres-sion. Annu Rev Pathol. 2010;5:99-118. google scholar
• Childs BG, Durik M, Baker DJ, van Deursen JM. Cellular senes-cence in aging and age-related disease: from mechanisms to ther-apy. Nat Med. 2015;21(12):1424-1435. google scholar
• Yuan J, Ofengeim D. A guide to cell death pathways. Nat Rev Mol Cell Biol. 2024;25(5):379-395. google scholar
• Han J, Zhong CQ, Zhang DW. Programmed necrosis: Backup to and competitor with apoptosis in the immune system. Nat Immunol. 2011;12(12):1143-1149. google scholar
• Sanz AB, Sanchez-Nino MD, Ramos AM, Ortiz A. Regu-lated cell death pathways in kidney disease. Nat Rev Nephrol. 2023;19(5):281-299. google scholar
• Franceschi C, Garagnani P, Parini P, Giuliani C, Santoro A. In-flammaging: A new immune-metabolic viewpoint for age-related diseases. Nat Rev Endocrinol. 2018;14(10):576-590. google scholar
• Ferrucci L, Fabbri E. Inflammageing: Chronic inflammation in ageing, cardiovascular disease, and frailty. Nat Rev Cardiol. 2018;15(9):505-522. google scholar
• Hotamisligil GS. Inflammation and metabolic disorders. Nature. 2006;444(7121):860-867. google scholar
• Salminen A, Kaarniranta K, Kauppinen A. Inflammaging: Disturbed interplay between autophagy and inflammasomes. Aging-Us. 2012;4(3):166-175. google scholar
• Esrefoglu M, Kalkan TK, Karatas E, et al. Hepatoprotective actions of melatonin by mainly modulating oxidative status and apoptosis rate in lipopolysaccharide-induced liver damage. Immunopharm Immunot. 2024;46(2):161-171. google scholar
• Lopez-Otm C, Blasco MA, Partridge L, Serrano M, Kroemer G. The hallmarks of aging. Cell. 2013;153(6):1194-1217. google scholar
• Mehdizadeh M, Aguilar M, Thorin E, Ferbeyre G, Nattel S. The role of cellular senescence in cardiac disease: basic biology and clinical relevance. Nat Rev Cardiol. 2022;19(4):250-264. google scholar
• Ishaq A, Tchkonia T, Kirkland JL, Siervo M, Saretzki G. Palmitate induces DNA damage and senescence in human adipocytes in vitro that can be alleviated by oleic acid but not inorganic nitrate. Exp Gerontol. 2022;163:111798. doi:10.1016/j.exger.2022.111798 google scholar
• Lagnado A, Leslie J, Ruchaud-Sparagano MH, et al. Neu-trophils induce paracrine telomere dysfunction and senes-cence in ROS-dependent manner. Embo J. 2021;40(9):e106048. doi:10.15252/embj.2020106048 google scholar
• Palmer AK, Gustafson B, Kirkland JL, Smith U. Cellular senes-cence: At the nexus between ageing and diabetes. Diabetologia. 2019;62(10):1835-1841. google scholar
• Miwa S, Kashyap S, Chini E, von Zglinicki T. Mitochon-drial dysfunction in cell senescence and aging. J Clin Invest. 2022;132(13):e158447. doi:10.1172/JCI158447 google scholar
• Xu M, Pirtskhalava T, Farr JN, et al. Senolytics improve physical function and increase lifespan in old age. Nat Med. 2018;24(8):1246-1256. google scholar
• Acosta JC, Banito A, Wuestefeld T, et al. A complex secretory program orchestrated by the inflammasome controls paracrine senescence. Nat Cell Biol. 2013;15(8):978-990. google scholar
• Franceschi C, Campisi J. Chronic inflammation (inflammag-ing) and its potential contribution to age-associated diseases. J Gerontol A Biol Sci Med Sci. 2014;69 Suppl 1:S4-S9. doi:10.1093/gerona/glu057 google scholar
• Stahl EC, Delgado ER, Alencastro F, et al. Inflammation and ectopic fat deposition in the aging murine liver is influenced by CCR2. Am J Pathol. 2020;190(2):372-387. google scholar
• Marquez-Exposito L, Tejedor-Santamaria L, Santos-Sanchez L, et al. Acute kidney injury is aggravated in aged mice by the exacerbation of proinflammatory processes. Front Pharmacol. 2021;12:662020. doi:10.3389/fphar.2021.662020 google scholar
• Tower J. Programmed cell death in aging. Ageing Res Rev. 2015;23(Pt A):90-100. google scholar
• Walczak H. TNF and ubiquitin at the crossroads of gene activation, cell death, inflammation, and cancer. Immunol Rev. 2011;244:9-28. google scholar
• Bertheloot D, Latz E, Franklin BS. Necroptosis, pyroptosis and apoptosis:An intricate game of cell death. Cell Mol Immunol. 2021;18(5):1106-1121. google scholar
• Vandenabeele P, Galluzzi L, Vanden Berghe T, Kroemer G. Molecular mechanisms of necroptosis: an ordered cellular ex-plosion. Nat Rev Mol Cell Biol. 2010;11(10):700-714. google scholar
• Newton K, Manning G. Necroptosis and inflammation. Annu Rev Biochem. 2016;85:743-763. google scholar
• Murphy JM, Czabotar PE, Hildebrand JM, et al. The pseudokinase MLKL mediates necroptosis via a molecular switch mechanism. Immunity. 2013;39(3):443-453. google scholar
• Sun L, Wang H, Wang Z, et al. Mixed lineage kinase domain-like protein mediates necrosis signaling downstream of RIP3 kinase. Cell. 2012;148(1-2):213-227. google scholar
• Silke J. The regulation of TNF signalling: What a tangled web we weave. Curr Opin Immunol. 2011;23(5):620-626. google scholar
• Kanayama A, Seth RB, Sun LJ, et al. TAB2 and TAB3 activate the NF-kB pathway through binding to polyubiquitin chains. Mol Cell. 2004;15(4):535-548. google scholar
• Mohammed S, Thadathil N, Selvarani R, et al. Necroptosis con-tributes to chronic inflammation and fibrosis in aging liver. Aging Cell. 2021;20(12):e13512. doi:10.1111/acel.13512 google scholar