Autophagy-related gene Rubicon deficiency inhibits cellular senescence in the heart

doi:10.12307/2022.573

Abstract

Abstract: BACKGROUND: Cardiac aging is the major risk factor for heart diseases. Cellular senescence is the key mechanism of cardiac aging, which plays an important role in age-related heart disease. Our previous work has shown that deletion of autophagy-related gene Uvrag promotes cellular senescence in the heart. Rubicon is an inhibitory interacting partner of Uvrag. However, the function of Rubicon in cellular senescence in the heart remains unknown.
OBJECTIVE: To determine the effect of Rubicon gene on cellular senescence in the heart.
METHODS: Wild type and Rubicon-deficient mice at 15 months of age were utilized. Uvrag-deficient mice were applied as observation objects. Uvrag-deficient mice of the same age were used as positive controls. Fluorescence quantitative PCR was used to detect changes in mRNA expression of senescence-related secreted phenotype-related factors in mouse heart tissue. Hematoxylin-eosin staining, Sirius red staining, and senescence-associated β-galactosidase staining were performed to observe myocardial histology. Western blot assay was conducted to measure the expression of p53 and p16 protein in the mouse heart. The protocols were approved by the Animal Experiment Ethics Committee of Shanghai Jiao Tong University.
RESULTS AND CONCLUSION: (1) Hematoxylin-eosin staining and Sirius red staining demonstrated that Rubicon deficiency ameliorated myocardial cell remodeling and cardiac fibrosis during aging. (2) Senescence-associated β-galactosidase staining displayed that Rubicon-deficient mice had significantly fewer senescence-associated β-galactosidase-positive cells in hearts from Rubicon-deficient mice than in control mice (P < 0.05). (3) Fluorescence quantitative PCR results suggested that expression of interleukin 1β, interleukin 6, transforming growth factor β, type III collagen α1 chain, and tissue inhibitor of matrix metalloproteinase 1 mRNA in hearts was significantly decreased in Rubicon-deficient mice than that in control mice (P < 0.05). (4) Western blot assay results showed that p53 protein abundance, a key protein regulating cellular senescence, was significantly lower in hearts from Rubicon-deficient mice than in control mice (P < 0.05). (5) It is indicated that Rubicon deficiency obviously inhibits cellular senescence in the heart and delays cardiac aging. Rubicon is a potential target for the treatment of cardiac aging and age-related heart disease.

Key words: Rubicon, cellular senescence, cardiac aging, senescence-associated secretory phenotype, p53, mice

CLC Number:

He Fan, Lai Shuaiwei, Zhang Shasha, Dong Fan, Amber Naz, Haniya Mazhar, He Lin, Zhu Hongxin. Autophagy-related gene Rubicon deficiency inhibits cellular senescence in the heart[J]. Chinese Journal of Tissue Engineering Research, 2022, 26(24): 3897-3902.

Figures/Tables 6

References

[1] FLORA GD, NAYAK MK. A Brief Review of Cardiovascular Diseases, Associated Risk Factors and Current Treatment Regimes. Curr Pharm Des. 2019;25(38):4063-4084.
[2] SHAKERI H, LEMMENS K, GEVAERT AB, et al. Cellular Senescence Links Aging and Diabetes in Cardiovascular Disease. Am J Physiol Heart Circ Physiol. 2018;315(3): H448-H462.
[3] CALCINOTTO A, KOHLI J, ZAGATO E, et al. Cellular Senescence: Aging, Cancer, and Injury. Physiol Rev. 2019;99(2):1047-1078.
[4] COLLADO M, SERRANO M. The Power and the Promise of Oncogene-induced Senescence Markers. Nat Rev Cancer. 2006;6(6):472-476.
[5] HERNANDEZ-SEGURA A, NEHME J, DEMARIA M. Hallmarks of Cellular Senescence. Trends Cell Biol. 2018;28(6):436-453.
[6] MIJIT M, CARACCIOLO V, MELILLO A, et al. Role of P53 in the Regulation of Cellular Senescence. Biomolecules. 2020;10(3):420.
[7] ZHU P, ZHANG C, GAO Y, et al. The Transcription Factor Slug Represses P16(Ink4a) and Regulates Murine Muscle Stem Cell Aging. Nat Commun. 2019;10(1):2568.
[8] ZHU Y, TCHKONIA T, PIRTSKHALAVA T, et al. The Achilles’ Heel of Senescent Cells: from Transcriptome to Senolytic Drugs. Aging Cell. 2015;14(4):644-658.
[9] WALASZCZYK A, DOOKUN E,REDGRAVE R, et al. Pharmacological Clearance of Senescent Cells Improves Survival and Recovery in Aged Mice Following Acute Myocardial Infarction. Aging Cell. 2019;18(3):e12945.
[10] SHIMIZU I, MINAMINO T. Cellular Senescence in Cardiac Diseases. J Cardiol. 2019; 74(4):313-319.
[11] WONG SQ, KUMAR AV, MILLS J, et al. Autophagy in Aging and Longevity. Hum Genet. 2020;139(3):277-290.
[12] GUO F,LIU X,CAI H, et al. Autophagy in Neurodegenerative Diseases:Pathogenesis and Therapy. Brain Pathol. 2018;28(1):3-13.
[13] LUO F, SANDHU AF, RUNGRATANAWANICH W, et al. Melatonin and Autophagy in Aging-Related Neurodegenerative Diseases. Int J Mol Sci. 2020;21(19):7174.
[14] ALLAIRE M, RAUTOU PE, CODOGNO P, et al. Autophagy in Liver Diseases: Time for Translation? J Hepatol. 2019;70(5):985-998.
[15] KE PY. Mitophagy in the Pathogenesis of Liver Diseases. Cells. 2020;9(4):831.
[16] DU J, LIU Y, FU J. Autophagy and Heart Failure. Adv Exp Med Biol. 2020;1207:223-227.
[17] CORSETTI G, PASINI E, ROMANO C, et al. How Can Malnutrition Affect Autophagy in Chronic Heart Failure? Focus and Perspectives. Int J Mol Sci. 2021;22(7):3332.
[18] YOUNG A, NARITA R, FERREIRA M, et al. Autophagy Mediates the Mitotic Senescence Transition. Genes Dev. 2009;23(7):798-803.
[19] KWON Y, KIM JW, JEOUNG JA, et al. Autophagy Is Pro-Senescence When Seen in Close-Up, but Anti-Senescence in Long-Shot. Mol Cells. 2017;40(9):607-612.
[20] PULAKAT L, CHEN HH. Pro-Senescence and Anti-Senescence Mechanisms of Cardiovascular Aging: Cardiac MicroRNA Regulation of Longevity Drug-Induced Autophagy. Front Pharmacol. 2020;11:774.
[21] ZHONG Y, WANG QJ, LI XT, et al. Distinct Regulation of Autophagic Activity by Atg14L and Rubicon Associated with Beclin 1-phosphatidylinositol-3-kinase Complex. Nature Cell Biol. 2009;11(4):468-476.
[22] MATSUNAGA K, SAITOH T, TABATA K, et al. Two Beclin 1-binding Proteins, Atg14L and Rubicon,Reciprocally Regulate Autophagy at Different Stages. Nat Cell Biol. 2009;11(4):385-396.
[23] KANG R, ZEH HJ, LOTZE MT, et al. The Beclin 1 Network Regulates Autophagy and Apoptosis. Cell Death and Differentiation. 2011;18(4):571-580.
[24] SUN, QM, ZHANG J, FAN, WL, et al. The RUN Domain of Rubicon Is Important for hVps34 Binding, Lipid Kinase Inhibition, and Autophagy Suppression. Journal of Biological Chemistry. 2011;286(1):185-191.
[25] SUN QM, ZHANG J, FAN WL, et al. Rubicon Controls Endosome Maturation as A Rab7 Effector. Proc Natl Acad Sci U S A. 2010;107(45):19338-19343.
[26] NAKAMURA S, OBA M, SUZUKI M, et al. Suppression of Autophagic Activity by Rubicon is A Signature of Aging. Nat Commun. 2019;10(1):847.
[27] SONG Z, AN L, YE Y, et al. Essential Role for UVRAG in Autophagy and Maintenance of Cardiac Function. Cardiovasc Res. 2014;101(1):48-56.
[28] 来帅威,张沙沙,刘晓芸,等. 心脏细胞衰老与Uvrag基因的缺失[J]. 中国组织工程研究,2021,25(14):2241-2246.
[29] ZI Z, SONG Z, ZHANG S, et al. Rubicon Deficiency Enhances Cardiac Autophagy and Protects Mice from Lipopolysaccharide-Induced Lethality and Reduction in Stroke Volume. J Cardiovasc Pharmacol. 2015;65(3):252-261.
[30] THOMAS N, GURVICH C, KULKARNI J. Sex Differences in Aging and Associated Biomarkers. Adv Exp Med Biol. 2019;1178:57-76.
[31] DODIG S, CEPELAK I, PAVIC I. Hallmarks of Senescence and Aging. Biochem Med (Zagreb). 2019;29(3):030501.
[32] ZHANG J, CHENG P, PU K. Recent Advances of Molecular Optical Probes in Imaging of beta-Galactosidase. Bioconjug Chem. 2019;30(8):2089-2101.
[33] LOPES-PACIENCIA S, SAINT-GERMAIN E, ROWELL MC, et al. The Senescence-associated Secretory Phenotype and its Regulation. Cytokine. 2019;117:15-22.
[34] HOSHINO A, MITA Y, OKAWA Y, et al. Cytosolic P53 Inhibits Parkin-Mediated Mitophagy and Promotes Mitochondrial Dysfunction in The Mouse Heart. Nat Commun. 2013;4:2308.
[35] REN X, CHEN L, XIE J, et al. Resveratrol Ameliorates Mitochondrial Elongation via Drp1/Parkin/PINK1 Signaling in Senescent-Like Cardiomyocytes. Oxid Med Cell Longev. 2017;2017:4175353.
[36] MADEO F, TAVERNARAKIS N, KROEMER G. Can autophagy promote longevity? Nat Cell Biol. 2010;12(9):842-846.
[37] FERNÁNDEZ ÁF, SEBTI S, WEI Y, et al. Disruption of The Beclin 1-BCL2 Autophagy Regulatory Complex Promotes Longevity in Mice. Nature. 2018;558(7708):136-140.
[38] XU S, CAI Y, WEI Y. mTOR Signaling from Cellular Senescence to Organismal Aging. Aging Dis. 2014;5(4):263-273.
[39] NISHIMURA A, SHIMAUCHI T, TANAKA T, et al. Hypoxia-Induced Interaction of Filamin with Drp1 Causes Mitochondrial Hyperfission-Associated Myocardial Senescence. Sci Signal. 2018;11(556):eaat5185.
[40] ANDERSON R, LAGNADO A, MAGGIORANI D, et al. Length-Independent Telomere Damage Drives Post-Mitotic Cardiomyocyte Senescence. EMBO J. 2019;38(5): e100492.
[41] LIU XY, ZHANG S, AN L, et al. Loss of Rubicon Ameliorates Doxorubicin-Induced Cardiotoxicity through Enhancement of Mitochondrial Quality. Int J Cardiol. 2019; 296:129-135.