Abstract
Spermidine (SPD) is a natural polyamine present in all living organisms and is involved in the maintenance of cellular homeostasis by inducing autophagy in different model organisms. Its role as a caloric restriction mimetic (CRM) is still being investigated. We have undertaken this study to investigate whether SPD, acting as a CRM, can confer neuroprotection in d-galactose induced accelerated senescence model rat and naturally aged rats through modulation of autophagy and inflammation. Young male rats (4 months), d-gal induced (500 mg/kg b.w., subcutaneously) aging and naturally aged (22 months) male rats were supplemented with SPD (10 mg/kg b.w., orally) for 6 weeks. Standard protocols were employed to measure prooxidants, antioxidants, apoptotic cell death and electron transport chain complexes in brain tissues. Gene expression analysis with reverse transcriptase-polymerase chain reaction (RT-PCR) was performed to assess the expression of autophagy and inflammatory marker genes. Our data demonstrate that SPD significantly (p ≤ 0.05) decreased the level of pro-oxidants and increased the level of antioxidants. SPD supplementation also augmented the activities of electron transport chain complexes in aged brain mitochondria thus proving its antioxidant potential at the level of mitochondria. RT-PCR data revealed that SPD up-regulated the expression of autophagy genes (ATG-3, Beclin-1, ULK-1 and LC3B) and down-regulated the expression of the inflammatory gene (IL-6) in aging brain. Our results provide first line of evidence that SPD provides neuroprotection against aging-induced oxidative stress by regulating autophagy, antioxidants level and also reduces neuroinflammation. These results suggest that SPD may be beneficial for neuroprotection during aging and age-related disorders.
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Andreazza AC, Shao L, Wang J-F, Young LT (2010) Mitochondrial complex I activity and oxidative damage to mitochondrial proteins in the prefrontal cortex of patients with bipolar disorder. Arch Gen Psychiatry 67:360–368. https://doi.org/10.1001/archgenpsychiatry.2010.22
Azman KF, Zakaria R (2019) d-Galactose-induced accelerated aging model: an overview. Biogerontology 20:763–782. https://doi.org/10.1007/s10522-019-09837-y
Bellé NAV, Dalmolin GD, Fonini G et al (2004) Polyamines reduces lipid peroxidation induced by different pro-oxidant agents. Brain Res 1008:245–251. https://doi.org/10.1016/j.brainres.2004.02.036
Benzie IFF, Strain JJ (1996) The ferric reducing ability of plasma (FRAP) as a measure of “antioxidant power”: the FRAP assay. Anal Biochem 239:70–76. https://doi.org/10.1006/abio.1996.0292
Bhukel A, Madeo F, Sigrist SJ (2017) Spermidine boosts autophagy to protect from synapse aging. Autophagy 13:444–445. https://doi.org/10.1080/15548627.2016.1265193
Budni J, Garcez ML, Mina F et al (2017) The oral administration of d-galactose induces abnormalities within the mitochondrial respiratory chain in the brain of rats. Metab Brain Dis 32:811–817. https://doi.org/10.1007/s11011-017-9972-9
Cakatay U, Telci A, Kayalì R et al (2001) Relation of oxidative protein damage and nitrotyrosine levels in the aging rat brain. Exp Gerontol 36:221–229
Castro MR, Suarez E, Kraiselburd E et al (2012) Aging increases mitochondrial DNA damage and oxidative stress in liver of rhesus monkeys. Exp Gerontol 47:29–37. https://doi.org/10.1016/j.exger.2011.10.002
Cebe T, Yanar K, Atukeren P et al (2014) A comprehensive study of myocardial redox homeostasis in naturally and mimetically aged rats. AGE. https://doi.org/10.1007/s11357-014-9728-y
Chen B, Zhong Y, Peng W et al (2010) Age-related changes in the central auditory system: comparison of d-galactose-induced aging rats and naturally aging rats. Brain Res 1344:43–53. https://doi.org/10.1016/j.brainres.2010.04.082
Cho KS, Lee JH, Cho J et al (2020) Autophagy Modulators and Neuroinflammation. Curr Med Chem 27:955–982. https://doi.org/10.2174/0929867325666181031144605
Cooper RL, Linnoila M (1977) Sexual behavior in aged, noncycling female rats. Physiol Behav 18:573–576. https://doi.org/10.1016/0031-9384(77)90054-3
Cui X, Zuo P, Zhang Q et al (2006) Chronic systemic d-galactose exposure induces memory loss, neurodegeneration, and oxidative damage in mice: protective effects of R-alpha-lipoic acid. J Neurosci Res 83:1584–1590. https://doi.org/10.1002/jnr.20845
Del Roso A, Vittorini S, Cavallini G et al (2003) Ageing-related changes in the in vivo function of rat liver macroautophagy and proteolysis. Exp Gerontol 38:519–527
Egan DF, Shackelford DB, Mihaylova MM et al (2011) Phosphorylation of ULK1 (hATG1) by AMP-activated protein kinase connects energy sensing to mitophagy. Science 331:456–461. https://doi.org/10.1126/science.1196371
Eisenberg T, Knauer H, Schauer A et al (2009) Induction of autophagy by spermidine promotes longevity. Nat Cell Biol 11:1305–1314. https://doi.org/10.1038/ncb1975
Eisenberg T, Abdellatif M, Schroeder S et al (2016) Cardioprotection and lifespan extension by the natural polyamine spermidine. Nat Med 22:1428–1438. https://doi.org/10.1038/nm.4222
Evans P, Lyras L, Halliwell B (1999) Measurement of protein carbonyls in human brain tissue. Methods Enzymol 300:145–156
Gabandé-Rodríguez E, de Las G, Heras MM, Mittelbrunn M (2019) Control of inflammation by calorie restriction mimetics: on the crossroad of autophagy and mitochondria. Cells. https://doi.org/10.3390/cells9010082
Galluzzi L, Bravo-San Pedro JM, Kroemer G (2016) Autophagy mediates tumor suppression via cellular senescence. Trends Cell Biol 26:1–3. https://doi.org/10.1016/j.tcb.2015.11.001
Garg G, Singh S, Singh AK, Rizvi SI (2017) Metformin alleviates altered erythrocyte redox status during aging in rats. Rejuvenation Res 20:15–24. https://doi.org/10.1089/rej.2016.1826
Green LC, Wagner DA, Glogowski J et al (1982) Analysis of nitrate, nitrite, and [15N]nitrate in biological fluids. Anal Biochem 126:131–138
Green DR, Galluzzi L, Kroemer G (2011) Mitochondria and the autophagy-inflammation-cell death axis in organismal aging. Science 333:1109–1112. https://doi.org/10.1126/science.1201940
Haider S, Liaquat L, Shahzad S et al (2015) A high dose of short term exogenous d-galactose administration in young male rats produces symptoms simulating the natural aging process. Life Sci 124:110–119. https://doi.org/10.1016/j.lfs.2015.01.016
Holloszy JO, Fontana L (2007) Caloric restriction in humans. Exp Gerontol 42:709–712. https://doi.org/10.1016/j.exger.2007.03.009
Hong I-S, Lee H-Y, Kim H-P (2014) Anti-oxidative effects of Rooibos tea (Aspalathus linearis) on immobilization-induced oxidative stress in rat brain. PLoS ONE 9:e87061. https://doi.org/10.1371/journal.pone.0087061
Igarashi K, Kashiwagi K (2010) Modulation of cellular function by polyamines. Int J Biochem Cell Biol 42:39–51. https://doi.org/10.1016/j.biocel.2009.07.009
Ingram DK, Roth GS (2015) Calorie restriction mimetics: can you have your cake and eat it, too? Ageing Res Rev 20:46–62. https://doi.org/10.1016/j.arr.2014.11.005
Jaeger PA, Wyss-Coray T (2010) Beclin 1 complex in autophagy and Alzheimer disease. Arch Neurol. https://doi.org/10.1001/archneurol.2010.258
Jamwal S, Kumar P (2016) Spermidine ameliorates 3-nitropropionic acid (3-NP)-induced striatal toxicity: Possible role of oxidative stress, neuroinflammation, and neurotransmitters. Physiol Behav 155:180–187. https://doi.org/10.1016/j.physbeh.2015.12.015
Jing Y-H, Yan J-L, Wang Q-J et al (2018) Spermidine ameliorates the neuronal aging by improving the mitochondrial function in vitro. Exp Gerontol 108:77–86. https://doi.org/10.1016/j.exger.2018.04.005
Kakkar P, Das B, Viswanathan PN (1984) A modified spectrophotometric assay of superoxide dismutase. Indian J Biochem Biophys 21:130–132
Kapoor N, Pant AB, Dhawan A et al (2006) Differences in sensitivity of cultured rat brain neuronal and glial cytochrome P450 2E1 to ethanol. Life Sci 79:1514–1522. https://doi.org/10.1016/j.lfs.2006.04.023
Kasote DM, Hegde MV, Katyare SS (2013) Mitochondrial dysfunction in psychiatric and neurological diseases: cause(s), consequence(s), and implications of antioxidant therapy. BioFactors Oxf Engl 39:392–406. https://doi.org/10.1002/biof.1093
Kiriyama Y, Nochi H (2015) The function of autophagy in neurodegenerative diseases. Int J Mol Sci 16:26797–26812. https://doi.org/10.3390/ijms161125990
Kou X, Li J, Liu X et al (1985) (2017) Swimming attenuates d-galactose-induced brain aging via suppressing miR-34a-mediated autophagy impairment and abnormal mitochondrial dynamics. J Appl Physiol Bethesda Md 122:1462–1469. https://doi.org/10.1152/japplphysiol.00018.2017
Kumar A, Prakash A, Dogra S (2010) Naringin alleviates cognitive impairment, mitochondrial dysfunction and oxidative stress induced by d-galactose in mice. Food Chem Toxicol 48:626–632. https://doi.org/10.1016/j.fct.2009.11.043
Le Noci V, Sommariva M, Bianchi F et al (2019) Local administration of caloric restriction mimetics to promote the immune control of lung metastases. J Immunol Res 2019:2015892. https://doi.org/10.1155/2019/2015892
Lei M, Hua X, Xiao M et al (2008) Impairments of astrocytes are involved in the d-galactose-induced brain aging. Biochem Biophys Res Commun 369:1082–1087. https://doi.org/10.1016/j.bbrc.2008.02.151
Leidal AM, Levine B, Debnath J (2018) Autophagy and the cell biology of age-related disease. Nat Cell Biol 20:1338–1348. https://doi.org/10.1038/s41556-018-0235-8
Li G, Ding H, Yu X et al (2020) Spermidine suppresses inflammatory DC function by activating the FOXO3 pathway and counteracts autoimmunity. iScience 23:100807. https://doi.org/10.1016/j.isci.2019.100807
Liang Y, Liu C, Lu M et al (2018) Calorie restriction is the most reasonable anti-ageing intervention: a meta-analysis of survival curves. Sci Rep. https://doi.org/10.1038/s41598-018-24146-z
Liapi C, Stolakis V, Zarros A et al (2013) Gestational exposure to cadmium alters crucial offspring rat brain enzyme activities: the role of cadmium-free lactation. Environ Toxicol Pharmacol 36:835–839. https://doi.org/10.1016/j.etap.2013.07.014
Liu H, Dong J, Song S et al (2019) Spermidine ameliorates liver ischaemia-reperfusion injury through the regulation of autophagy by the AMPK-mTOR-ULK1 signalling pathway. Biochem Biophys Res Commun 519:227–233. https://doi.org/10.1016/j.bbrc.2019.08.162
Long J, Wang X, Gao H et al (2007) d-galactose toxicity in mice is associated with mitochondrial dysfunction: protecting effects of mitochondrial nutrient R-alpha-lipoic acid. Biogerontology 8:373–381. https://doi.org/10.1007/s10522-007-9081-y
López-Otín C, Blasco MA, Partridge L et al (2013) The hallmarks of aging. Cell 153:1194–1217. https://doi.org/10.1016/j.cell.2013.05.039
Madeo F, Pietrocola F, Eisenberg T, Kroemer G (2014) Caloric restriction mimetics: towards a molecular definition. Nat Rev Drug Discov 13:727–740. https://doi.org/10.1038/nrd4391
Madeo F, Eisenberg T, Pietrocola F, Kroemer G (2018) Spermidine in health and disease. Science. https://doi.org/10.1126/science.aan2788
Mariño G, Pietrocola F, Madeo F, Kroemer G (2014) Caloric restriction mimetics: natural/physiological pharmacological autophagy inducers. Autophagy 10:1879–1882. https://doi.org/10.4161/auto.36413
Martins R, Lithgow GJ, Link W (2016) Long live FOXO: unraveling the role of FOXO proteins in aging and longevity. Aging Cell 15:196–207. https://doi.org/10.1111/acel.12427
Morselli E, Galluzzi L, Kepp O, et al (2009) Autophagy mediates pharmacological lifespan extension by spermidine and resveratrol. Aging 1:961–970. https://doi.org/10.18632/aging.100110
Nakamura S, Yoshimori T (2018) Autophagy and longevity. Mol Cells 41:65–72. https://doi.org/10.14348/molcells.2018.2333
Nakamura S, Oba M, Suzuki M et al (2019) Suppression of autophagic activity by Rubicon is a signature of aging. Nat Commun 10:847. https://doi.org/10.1038/s41467-019-08729-6
Navarro A, Gómez C, Sánchez-Pino M-J et al (2005) Vitamin E at high doses improves survival, neurological performance, and brain mitochondrial function in aging male mice. Am J Physiol Regul Integr Comp Physiol 289:R1392–1399. https://doi.org/10.1152/ajpregu.00834.2004
Omata Y, Lim Y-M, Akao Y, Tsuda L (2014) Age-induced reduction of autophagy-related gene expression is associated with onset of Alzheimer’s disease. Am J Neurodegener Dis 3:134–142
Parameshwaran K, Irwin MH, Steliou K, Pinkert CA (2010) d-galactose effectiveness in modeling aging and therapeutic antioxidant treatment in mice. Rejuvenation Res 13:729–735. https://doi.org/10.1089/rej.2010.1020
Parihar MS, Nazarewicz RR, Kincaid E et al (2008) Association of mitochondrial nitric oxide synthase activity with respiratory chain complex I. Biochem Biophys Res Commun 366:23–28. https://doi.org/10.1016/j.bbrc.2007.11.056
Pierzynowska K, Gaffke L, Cyske Z et al (2018) Autophagy stimulation as a promising approach in treatment of neurodegenerative diseases. Metab Brain Dis 33:989–1008. https://doi.org/10.1007/s11011-018-0214-6
Qi Y, Qiu Q, Gu X et al (2016) ATM mediates spermidine-induced mitophagy via PINK1 and Parkin regulation in human fibroblasts. Sci Rep 6:24700. https://doi.org/10.1038/srep24700
Qun VEJ, Moghaddas S et al (2003) Production of reactive oxygen species by mitochondria: central role of complex III. J Biol Chem 278:36027–36031. https://doi.org/10.1074/jbc.M304854200
Rider JE, Hacker A, Mackintosh CA et al (2007) Spermine and spermidine mediate protection against oxidative damage caused by hydrogen peroxide. Amino Acids 33:231–240. https://doi.org/10.1007/s00726-007-0513-4
Ross KS, Smith C (2020) d-galactose: a model of accelerated ageing sufficiently sensitive to reflect preventative efficacy of an antioxidant treatment. Biogerontology. https://doi.org/10.1007/s10522-020-09891-x
Rubinsztein DC, Mariño G, Kroemer G (2011) Autophagy and aging. Cell 146:682–695. https://doi.org/10.1016/j.cell.2011.07.030
Sastre J, Pallardó FV, García de la Asunción J, Viña J (2000) Mitochondria, oxidative stress and aging. Free Radic Res 32:189–198. https://doi.org/10.1080/10715760000300201
Sedlak J, Lindsay RH (1968) Estimation of total, protein-bound, and nonprotein sulfhydryl groups in tissue with Ellman’s reagent. Anal Biochem 25:192–205
Sharma S, Kumar P, Deshmukh R (2018) Neuroprotective potential of spermidine against rotenone induced Parkinson’s disease in rats. Neurochem Int 116:104–111. https://doi.org/10.1016/j.neuint.2018.02.010
Shwe T, Pratchayasakul W, Chattipakorn N, Chattipakorn SC (2018) Role of d-galactose-induced brain aging and its potential used for therapeutic interventions. Exp Gerontol 101:13–36. https://doi.org/10.1016/j.exger.2017.10.029
Simsek B, Yanar K, Kansu AD et al (2019) Caloric restriction improves the redox homeostasis in the aging male rat heart even when started in middle-adulthood and when the body weight is stable. Biogerontology 20:127–140. https://doi.org/10.1007/s10522-018-9781-5
Singh AK, Bissoyi A, Kashyap MP et al (2017) Autophagy activation alleviates amyloid-β-induced oxidative stress, apoptosis and neurotoxicity in human neuroblastoma SH-SY5Y cells. Neurotox Res. https://doi.org/10.1007/s12640-017-9746-5
Singh S, Singh AK, Garg G, Rizvi SI (2018) Fisetin as a caloric restriction mimetic protects rat brain against aging induced oxidative stress, apoptosis and neurodegeneration. Life Sci 193:171–179. https://doi.org/10.1016/j.lfs.2017.11.004
Singh AK, Singh S, Tripathi VK et al (2019) Rapamycin confers neuroprotection against aging-induced oxidative stress, mitochondrial dysfunction, and neurodegeneration in old rats through activation of autophagy. Rejuvenation Res 22:60–70. https://doi.org/10.1089/rej.2018.2070
Soda K, Dobashi Y, Kano Y et al (2009) Polyamine-rich food decreases age-associated pathology and mortality in aged mice. Exp Gerontol 44:727–732. https://doi.org/10.1016/j.exger.2009.08.013
Soto-Heredero G, Baixauli F, Mittelbrunn M (2017) Interorganelle communication between mitochondria and the endolysosomal system. Front Cell Dev Biol 5:95. https://doi.org/10.3389/fcell.2017.00095
Srividhya R, Zarkovic K, Stroser M et al (2009) Mitochondrial alterations in aging rat brain: effective role of (−)-epigallo catechin gallate. Int J Dev Neurosci 27:223–231. https://doi.org/10.1016/j.ijdevneu.2009.01.003
Su Y, Sun H, Fang J et al (2010) Brain mitochondrial dysfunction in ovariectomized mice injected with d-galactose. Neurochem Res 35:399–404. https://doi.org/10.1007/s11064-009-0068-8
Sun N, Youle RJ, Finkel T (2016) The mitochondrial basis of aging. Mol Cell 61:654–666. https://doi.org/10.1016/j.molcel.2016.01.028
von Kobbe C (2018) Cellular senescence: a view throughout organismal life. Cell Mol Life Sci 75:3553–3567. https://doi.org/10.1007/s00018-018-2879-8
Webb AE, Brunet A (2014) FOXO transcription factors: key regulators of cellular quality control. Trends Biochem Sci 39:159–169. https://doi.org/10.1016/j.tibs.2014.02.003
Wirawan E, Berghe TV, Lippens S et al (2012) Autophagy: for better or for worse. Cell Res 22:43–61. https://doi.org/10.1038/cr.2011.152
Wirth M, Benson G, Schwarz C et al (2018) The effect of spermidine on memory performance in older adults at risk for dementia: a randomized controlled trial. Cortex J Devoted Study Nerv Syst Behav 109:181–188. https://doi.org/10.1016/j.cortex.2018.09.014
Xu T-T, Li H, Dai Z, et al (2020) Spermidine and spermine delay brain aging by inducing autophagy in SAMP8 mice. Aging 12:6401–6414. https://doi.org/10.18632/aging.103035
Yanar K, Aydın S, Cakatay U et al (2011) Protein and DNA oxidation in different anatomic regions of rat brain in a mimetic ageing model. Basic Clin Pharmacol Toxicol 109:423–433. https://doi.org/10.1111/j.1742-7843.2011.00756.x
Yanar K, Simsek B, Atukeren P et al (2019a) Is d-Galactose a useful agent for accelerated aging model of gastrocnemius and soleus muscle of Sprague-Dawley rats? Rejuvenation Res 22:521–528. https://doi.org/10.1089/rej.2019.2185
Yanar K, Simsek B, Çaylı N et al (2019b) Caloric restriction and redox homeostasis in various regions of aging male rat brain: Is caloric restriction still worth trying even after early-adulthood?: Redox homeostasis and caloric restriction in brain. J Food Biochem 43:e12740. https://doi.org/10.1111/jfbc.12740
Acknowledgements
This work was also supported by a research grant to SIR from SERB-DST, Govt of India (EMR/2016/006470). Sandeep Singh is the recipient of a Senior Research Fellowship (3/1/2(3)/GER/2018-NCD-II) from Indian Council of Medical Research, India. The Department of Biochemistry is supported by FIST grant of DST, New Delhi, and SAP DRS I from UGC, India.
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Singh, S., Kumar, R., Garg, G. et al. Spermidine, a caloric restriction mimetic, provides neuroprotection against normal and d-galactose-induced oxidative stress and apoptosis through activation of autophagy in male rats during aging. Biogerontology 22, 35–47 (2021). https://doi.org/10.1007/s10522-020-09900-z
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DOI: https://doi.org/10.1007/s10522-020-09900-z