Xi-Kun Yuan Pin-Shi Ni Zhen-Hao Yan Zhi Yu Zhuang-Zhi Wang Chen-Kai Zhang Fang-Hui Li Xiao-Ming Yu 1Sports Department, Nanjing University of Science and Technology ZiJin College, Nanjing, China, 2School of Sport Sciences, Nanjing Normal University, Nanjing, China, 3Shanghai Seventh People's Hospital, Shanghai, China To investigate the effects of life-long exercise (LLE) on age-related inflammatory cytokines, apoptosis, oxidative stress, ferroptosis markers, and the NRF2/KAEP 1/Klotho pathway in rats. Eight-month-old female Sprague-Dawley rats were divided into four groups: 1) LLE: 18-month LLE training starting at 8 months of age, 2) Old moderate-intensity continuous training (OMICT): 8 months of moderate-intensity continuous training starting at 18 months of age, 3) Adult sedentary (ASED): 8 month-old adult sedentary control group, and 4) Old sedentary (OSED): a 26-month-old sedentary control group. Hematoxylin eosin staining was performed to observe the pathological changes of kidney tissue injury in rats; Masson's staining to observe the deposition of collagen fibers in rat kidney tissues; and western blotting to detect the expression levels of IL-6, IL 1beta, p53, p21, TNF-alpha, GPX4, KAEP 1, NRF2, SLC7A11, and other proteins in kidney tissues. Results: Compared with the ASED group, the OSED group showed significant morphological changes in renal tubules and glomeruli, which were swollen and deformed, with a small number of inflammatory cells infiltrated in the tubules. Compared with the OSED group, the expression levels of inflammation-related proteins such as IL-1beta, IL-6, TNF alpha, and MMP3 were significantly lower in the LLE group. Quantitative immunofluorescence analysis and western blotting revealed that compared with the ASED group, KAEP 1 protein fluorescence intensity and protein expression levels were significantly enhanced, while Klotho and NRF2 protein fluorescence intensity and protein expression levels were reduced in the OSED group. Compared with the OSED group, KAEP 1 protein fluorescence intensity and protein expression levels were reduced in the LLE and OMICT groups. Klotho and KAEP 1 protein expression levels and immunofluorescence intensity were higher in the LLE group than in the OSED group. The expression levels of GPX4 and SLC7A11, two negative marker proteins associated with ferroptosis, were significantly higher in the LLE group than in the OSED group, while the expression of p53 a cellular senescence-associated protein that negatively regulates SLC7A11, and the downstream protein p21 were significantly decreased. LLE may ameliorated aging-induced oxidative stress, inflammatory response, apoptosis, and ferroptosis by regulating Klotho and synergistically activating the NRF2/KAEP 1 pathway. Keywords: Life-long exercise, Moderate intensity continuous training, Aging, Kidney tissue, Ferroptosis.

Shanghai Seventh People's Hospital Shanghai China College of Sports Science Nanjing Normal University Nanjing China

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Kimura T, Isaka Y, Yoshimori T. Autophagy and kidney inflammation. Autophagy. 2017 Jun 3;13(6):997–1003. doi: 10.1080/15548627.2017.1309485. PubMed DOI PMC

Zhou XJ, Rakheja D, Yu X, Saxena R, Vaziri ND, Silva FG. The aging kidney. Kidney Int. 2008 Sep;74(6):710–20. PubMed

Zeisberg Michael, Neilson Eric G. Mechanisms of tubulointerstitial fibrosis. JASN. 2010;21(11):1819–34. doi: 10.1681/ASN.2010080793. PubMed DOI

Jung KJ, Min KJ, Park JW, Park KM, Kwon TK. Carnosic acid attenuates unilateral ureteral obstruction-induced kidney fibrosis via inhibition of Akt-mediated Nox4 expression. Free Radic Biol Med. 2016 Aug;97:50–57. doi: 10.1016/j.freeradbiomed.2016.05.020. PubMed DOI

Denic A, Glassock RJ, Rule AD. Structural and functional changes with the aging kidney. Adv Chronic Kidney Dis. 2016 Jan;23(1):19–28. doi: 10.1053/j.ackd.2015.08.004. PubMed DOI PMC

Hans-Joachim Anders, Schaefer Liliana. Beyond tissue injury-damage-associated molecular patterns, toll-like receptors, and inflammasomes also drive regeneration and fibrosis. J Am Soc Nephrol: JASN. 2014;25(7):1387–400. doi: 10.1681/ASN.2014010117. PubMed DOI PMC

Baar MP, Brandt RMC, Putavet DA, Klein JDD, Derks KWJ, Bourgeois BRM, Stryeck S, et al. Targeted apoptosis of senescent cells restores tissue homeostasis in response to chemotoxicity and aging. Cell. 2017;169(1):132–147.e16. doi: 10.1016/j.cell.2017.02.031. PubMed DOI PMC

Tower J. Programmed cell death in aging. Ageing Research Reviews. 2015;23(Part A):90–100. doi: 10.1016/j.arr.2015.04.002. PubMed DOI PMC

Balaban RS, Nemoto S, Finkel T. Mitochondria, oxidants, and aging. Cell. 2005 Feb 25;120(4):483–95. doi: 10.1016/j.cell.2005.02.001. PubMed DOI

Takahashi A, Kimura T, Takabatake Y, Namba T, Kaimori J, Kitamura H, Matsui I, et al. Autophagy guards against cisplatin-induced acute kidney injury. Am J Pathol. 2012;180(2):517–25. doi: 10.1016/j.ajpath.2011.11.001. PubMed DOI

Jiang X, Stockwell BR, Conrad M. Ferroptosis: mechanisms, biology and role in disease. Nat Rev Mol Cell Biol. 2021 Apr;22(4):266–282. doi: 10.1038/s41580-020-00324-8. PubMed DOI PMC

Wang J, Liu Y, Wang Y, Sun L. The cross-link between ferroptosis and kidney diseases. Oxid Med Cell Longev. 2021 May 3;2021:6654887. doi: 10.1155/2021/6654887. PubMed DOI PMC

Mora JC, Valencia WM. Exercise and Older Adults. Clin Geriatr Med. 2018;34(1):145–162. doi: 10.1016/j.cger.2017.08.007. PubMed DOI

Ruegsegger GN, Booth FW. Health benefits of exercise. Cold Spring Harb Perspect Med. 2018;8(7):a029694. doi: 10.1101/cshperspect.a029694. PubMed DOI PMC

Wang HT, Yang WQ, Liu YQ. Effects of aerobic exercise on Nrf2/GPX4/Ferroptosis pathway in myocardial injury in high-fat diet mice (in Chinese) Zhongguo Ying Yong Sheng Li Xue Za Zhi. 2022;38(2):143–148. PubMed

Padilha CS, Figueiredo C, Minuzzi LG, Chimin P, Deminice R, Krüger K, Rosa-Neto JC, et al. Immunometabolic responses according to physical fitness status and lifelong exercise during aging: New roads for exercise immunology. Ageing Res Rev. 2021;68:101341. doi: 10.1016/j.arr.2021.101341. PubMed DOI

Bellezza I, Giambanco I, Minelli A, Donato R. Nrf2-Keap1 signaling in oxidative and reductive stress. Biochim Biophys Acta Mol Cell Res. 2018;1865(5):721–733. doi: 10.1016/j.bbamcr.2018.02.010. PubMed DOI

Chen K, Wang S, Sun QW, Zhang B, Ullah M, Sun Z. Klotho Deficiency Causes Heart Aging via Impairing the Nrf2-GR Pathway. Circ Res. 2021;128(4):492–507. doi: 10.1161/CIRCRESAHA.120.317348. PubMed DOI PMC

Gao HE, Wu DS, Sun L, Yang LD, Qiao YB, Ma S, Wu ZJ, et al. Effects of lifelong exercise on age-related body composition, oxidative stress, inflammatory cytokines, and skeletal muscle proteome in rats. Mech Ageing Dev. 2020;189:111262. doi: 10.1016/j.mad.2020.111262. PubMed DOI

Li FH, Sun L, Wu DS, Gao HE, Min Z. Proteomics-based identification of different training adaptations of aged skeletal muscle following long-term high-intensity interval and moderate-intensity continuous training in aged rats. Aging (Albany NY) 2019;11(12):4159–4182. doi: 10.18632/aging.102044. PubMed DOI PMC

Wang WJ, Cai GY, Ning YC, Cui J, Hong Q, Bai XY, Xu XM, et al. Hydrogen sulfide mediates the protection of dietary restriction against renal senescence in aged F344 rats. Sci Rep. 2016;6:30292. doi: 10.1038/srep30292. PubMed DOI PMC

Kooman JP, van der Sande FM, Leunissen KM. Kidney disease and aging: A reciprocal relation. Exp Gerontol. 2017;87(Pt B):156–159. doi: 10.1016/j.exger.2016.02.003. PubMed DOI

Gao Q, Chen F, Zhang L, Wei A, Wang Y, Wu Z, Cao W. Inhibition of DNA methyltransferase aberrations reinstates antioxidant aging suppressors and ameliorates renal aging. Aging Cell. 2022;21(1):e13526. doi: 10.1111/acel.13526. PubMed DOI PMC

Chen Q, Cai J, Li X, Song A, Guo H, Sun Q, Yang C, Yang P. Progranulin promotes regeneration of inflammatory periodontal bone defect in rats via anti-inflammation, osteoclastogenic inhibition, and osteogenic promotion. Inflammation. 2019;42(1):221–234. doi: 10.1007/s10753-018-0886-4. PubMed DOI

Baker RG, Hayden MS, Ghosh S. NF-κB, inflammation, and metabolic disease. Cell Metab. 2011;13(1):11–22. doi: 10.1016/j.cmet.2010.12.008. PubMed DOI PMC

Sun L, Li FH, Li T, Min Z, Yang LD, Gao HE, Wu DS, et al. Effects of high-intensity interval training on adipose tissue lipolysis, inflammation, and metabolomics in aged rats. Pflugers Arch. 2020;472(2):245–258. doi: 10.1007/s00424-020-02351-y. PubMed DOI

Xu TT, Li H, Dai Z, Lau GK, Li BY, Zhu WL, Liu XQ, et al. Spermidine and spermine delay brain aging by inducing autophagy in SAMP8 mice. Aging (Albany NY) 2020;12(7):6401–6414. doi: 10.18632/aging.103035. PubMed DOI PMC

Lu J, Temp U, Müller-Hartmann A, Esser J, Grönke S, Partridge L. Sestrin is a key regulator of stem cell function and lifespan in response to dietary amino acids. Nat Aging. 2021;1(1):60–72. doi: 10.1038/s43587-020-00001-30. PubMed DOI

Su LJ, Zhang JH, Gomez H, Murugan R, Hong X, Xu D, Jiang F, et al. Reactive oxygen species-induced lipid peroxidation in apoptosis, autophagy, and ferroptosis. Oxid Med Cell Longev. 2019;2019:5080843. doi: 10.1155/2019/5080843. PubMed DOI PMC

Vucicevic K, Jakovljevic V, Colovic N, Tosic N, Kostic T, Glumac I, Pavlovic S, et al. Association of bax expression and bcl2/bax ratio with clinical and molecular prognostic markers in chronic lymphocytic leukemia. J Med Biochem. 2016;35(2):150–157. doi: 10.1515/jomb-2015-0017. PubMed DOI PMC

Zhi-Hui Huang, Liu Hao. Change of apoptosis level of brain and kidney in aged rats and the intervention effects of aerobic exercise. J Shandong Inst Physical Edu and Sports. (91) 2008;(08):47–50.

Done AJ, Traustadóttir T. Nrf2 mediates redox adaptations to exercise. Redox Biol. 2016;10:191–199. doi: 10.1016/j.redox.2016.10.003. PubMed DOI PMC

Alves FM, Ayton S, Bush AI, Lynch GS, Koopman R. Age-related changes in skeletal muscle iron homeostasis. J Gerontol A Biol Sci Med Sci. 2023 Jan 26;78(1):16–24. doi: 10.1093/gerona/glac139. PubMed DOI

Fathi R, Nasiri K, Akbari A, Ahmadi-KaniGolzar F, Farajtabar Z. Exercise protects against ethanol-induced damage in rat heart and liver through the inhibition of apoptosis and activation of NRF2/Keap-1/HO-1 pathway. Life Sci. 2020;256:117958. doi: 10.1016/j.lfs.2020.117958. PubMed DOI

Liu F, Wu S, Ren H, Gu J. Klotho suppresses RIG-I-mediated senescence-associated inflammation. Nat Cell Biol. 2011;13(3):254–62. doi: 10.1038/ncb2167. PubMed DOI

Angulo J, El Assar M, Rodríguez-Mañas L. Frailty and sarcopenia as the basis for the phenotypic manifestation of chronic diseases in older adults. Mol Aspects Med. 2016;50:1–32. doi: 10.1016/j.mam.2016.06.001. PubMed DOI

Guo Yue, Zou Jing, Wu Lingling, Zhuang Xiaodong, Liao Xinxue. Klotho Attenuates Hyperglycemia-Induced Cardiac Hypertrophy and Dysfunction by Concomitantly Activating Nrf2 and Brg1. Circulation. 2019;140:A14694. PubMed

Maltese G, Psefteli PM, Rizzo B, Srivastava S, Gnudi L, Mann GE, Siow RC. The anti-ageing hormone klotho induces Nrf2-mediated antioxidant defences in human aortic smooth muscle cells. J Cell Mol Med. 2017;21(3):621–627. doi: 10.1111/jcmm.12996. PubMed DOI PMC

Gao W, Guo L, Yang Y, Wang Y, Xia S, Gong H, Zhang BK, et al. Dissecting the crosstalk between NRF2 and NF-κB response pathways in drug-induced toxicity. Front Cell Dev Biol. 2022;9:809952. doi: 10.3389/fcell.2021.809952. PubMed DOI PMC

Bishop NC, Burton JO, Graham-Brown MPM, Stensel DJ, Viana JL, Watson EL. Exercise and chronic kidney disease: potential mechanisms underlying the physiological benefits. Nat Rev Nephrol. 2023;19(4):244–256. doi: 10.1038/s41581-022-00675-9. PubMed DOI

Shiyu Han. Research of relationship between kidney and aging from theory of “Man Eight and Woman Seven” in Suwen·Shanggu Tianzhen. Clin J Trad Chinese Med. 2017;29:76–78.

Dhillon RJ, Hasni S. Pathogenesis and management of sarcopenia. Clin Geriatr Med. 2017;33(1):17–26. doi: 10.1016/j.cger.2016.08.002. PubMed DOI PMC

Volpato S, Bianchi L, Cherubini A, Landi F, Maggio M, Savino E, Bandinelli S, et al. Prevalence and clinical correlates of sarcopenia in community-dwelling older people: application of the EWGSOP definition and diagnostic algorithm. J Gerontol A Biol Sci Med Sci. 2014;69(4):438–46. doi: 10.1093/gerona/glt149. PubMed DOI PMC

Kirchengast S, Huber J. Gender and age differences in lean soft tissue mass and sarcopenia among healthy elderly. Anthropol Anz. 2009;67(2):139–151. doi: 10.1127/0003-5548/2009/0018. PubMed DOI

Shin MJ, Jeon YK, Kim IJ. Testosterone and sarcopenia. World J Mens Health. 2018;36(3):192–198. https://doi.org/10.5534/wjmh.2012.30.3.192, https://doi.org/10.5534/wjmh.180001. PubMed DOI PMC

Wu R, Delahunt E, Ditroilo M, Lowery M, De Vito G. Effects of age and sex on neuromuscular-mechanical determinants of muscle strength. Age. 2016;38:57. doi: 10.1007/s11357-016-9921-2. PubMed DOI PMC

Bin You, Xiuzhu Huang, Yu Lin, Xi Huang. Changes in bone morphology, bone mineral density and bone metabolism in different parts of natural aging rats. Chinese J Tissue Eng Res. 2021;25:4118–4122. https://www.cjter.com/EN/Y2021/V25/I26/4118 .