SpringerLink
Log in

Impaired autophagic function in rat islets with aging

  • Published:
AGE Aims and scope Submit manuscript

Abstract

Type 2 diabetes is characterized by a deficit in β-cell function and mass, and its incidence increases with age. Autophagy is a highly regulated intracellular process for degrading cytoplasmic components, particularly protein aggregates and damaged organelles. Impaired or deficient autophagy is believed to cause or contribute to aging and age-related disease. Autophagy may be necessary to maintain structure, mass, and function of pancreatic β-cells. In this study, we investigated the effects of age on β-cell function and autophagy in pancreatic islets of 4-month-old (young), 14-month-old (adult), and 24-month-old (old) male Wistar rats. We found that islet β-cell function decreased gradually with age. Protein expression of the autophagy markers LC3/Atg8 and Atg7 exhibited a marked decline in aged islets. The expression of Lamp-2, a good indicator of autophagic degradation rate, was significantly reduced in the islets of old rats, suggesting that autophagic degradation is decreased in the islets of aged rats. However, protein expression of beclin-1/Atg6, which plays an important role in the induction and formation of the pre-autophagosome structure by associating with a multimeric complex of autophagy regulatory proteins (Atg14, Vps34/class 3 PI3 kinase, and Vps15), was most prominent in the islets of adult rats, and was higher in 24-month-old islets than in 4-month-old islets. The levels of p62/SQSTM1 and polyubiquitin aggregates, representing the functions of autophagy and proteasomal degradation, were increased in aging islets. 8-Hydroxydeoxyguanosine, a marker of mitochondrial and nuclear DNA oxidative damage, exhibited strong immunostaining in old islets. Analysis by electron microscopy demonstrated swelling and disintegration of cristae in the mitochondria of aged islets. These results suggest that β-cell and autophagic function in islets decline simultaneously with increasing age in Wistar rats, and that impaired autophagy in the islets of older rats may cause accumulation of misfolded and aggregated proteins and reduce the removal of abnormal mitochondria in β-cells, leading to reduced β-cell function. Dysfunctional autophagy in islets during the aging process may be an important mechanism leading to the development of type 2 diabetes.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Subscribe and save

Springer+ Basic
EUR 32.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or Ebook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Price includes VAT (United Kingdom)

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8
Fig. 9

Similar content being viewed by others

References

  • Angelopoulou R, Lavranos G, Manolakou P (2009) ROS in the aging male: model diseases with ROS-related pathophysiology. Reprod Toxicol 28:167–171

    Article  PubMed  CAS  Google Scholar 

  • Beckman KB, Ames BN (1998) The free radical theory of aging matures. Physiol Rev 78:547–581

    PubMed  CAS  Google Scholar 

  • Bjorkoy G, Lamark T, Brech A, Outzen H, Perander M, Overvatn A, Stenmark H, Johansen T (2005) p62/SQSTM1 forms protein aggregates degraded by autophagy and has a protective effect on huntingtin-induced cell death. J Cell Biol 171:603–614

    Article  PubMed  Google Scholar 

  • Bjorkoy G, Lamark T, Johansen T (2006) p62/SQSTM1: a missing link between protein aggregates and the autophagy machinery. Autophagy 2:138–139

    PubMed  Google Scholar 

  • Blagosklonny MV (2008) Aging: ROS or TOR. Cell Cycle 7:3344–3354

    Article  PubMed  CAS  Google Scholar 

  • Cadenas E, Davies KJ (2000) Mitochondrial free radical generation, oxidative stress, and aging. Free Radic Biol Med 29:222–230

    Article  PubMed  CAS  Google Scholar 

  • Cao Y, Klionsky DJ (2007) Physiological functions of Atg6/Beclin 1: a unique autophagy-related protein. Cell Res 17:839–849

    Article  PubMed  CAS  Google Scholar 

  • Cowie CC, Rust KF, Byrd-Holt DD, Eberhardt MS, Flegal KM, Engelgau MM, Saydah SH, Williams DE, Geiss LS, Gregg EW (2006) Prevalence of diabetes and impaired fasting glucose in adults in the U.S. population: National Health And Nutrition Examination Survey 1999–2002. Diabetes Care 29:1263–1268

    Article  PubMed  Google Scholar 

  • Cui J, Bai XY, Shi S, Cui S, Hong Q, Cai G, Chen X (2012) Age-related changes in the function of autophagy in rat kidneys. Age (Dordr) 34:329-339

    Google Scholar 

  • Dimri GP, Lee X, Basile G, Acosta M, Scott G, Roskelley C, Medrano EE, Linskens M, Rubelj I, Pereira-Smith O et al (1995) A biomarker that identifies senescent human cells in culture and in aging skin in vivo. Proc Natl Acad Sci U S A 92:9363–9367

    Article  PubMed  CAS  Google Scholar 

  • Drews G, Krippeit-Drews P, Dufer M (2010) Oxidative stress and beta-cell dysfunction. Pflugers Arch 460:703–718

    Article  PubMed  CAS  Google Scholar 

  • Ebato C, Uchida T, Arakawa M, Komatsu M, Ueno T, Komiya K, Azuma K, Hirose T, Tanaka K, Kominami E et al (2008) Autophagy is important in islet homeostasis and compensatory increase of beta cell mass in response to high-fat diet. Cell Metab 8:325–332

    Article  PubMed  CAS  Google Scholar 

  • Eskelinen EL (2006) Roles of LAMP-1 and LAMP-2 in lysosome biogenesis and autophagy. Mol Aspects Med 27:495–502

    Article  PubMed  CAS  Google Scholar 

  • Eskelinen EL, Illert AL, Tanaka Y, Schwarzmann G, Blanz J, Von Figura K, Saftig P (2002) Role of LAMP-2 in lysosome biogenesis and autophagy. Mol Biol Cell 13:3355–3368

    Article  PubMed  CAS  Google Scholar 

  • Gu Z, Du Y, Liu Y, Ma L, Li L, Gong Y, Tian H, Li C (2011) Effect of aging on islet beta-cell function and its mechanisms in Wistar rats. Age (Dordr). 7 Sept 2011. doi:10.1007/s11357-011-9312-7

  • He C, Klionsky DJ (2009) Regulation mechanisms and signaling pathways of autophagy. Annu Rev Genet 43:67–93

    Article  PubMed  CAS  Google Scholar 

  • Hur KY, Jung HS, Lee MS (2010) Role of autophagy in beta-cell function and mass. Diabetes Obes Metab 12(Suppl 2):20–26

    Article  PubMed  CAS  Google Scholar 

  • Ihm SH, Matsumoto I, Sawada T, Nakano M, Zhang HJ, Ansite JD, Sutherland DE, Hering BJ (2006) Effect of donor age on function of isolated human islets. Diabetes 55:1361–1368

    Article  PubMed  CAS  Google Scholar 

  • Ihm SH, Moon HJ, Kang JG, Park CY, Oh KW, Jeong IK, Oh YS, Park SW (2007) Effect of aging on insulin secretory function and expression of beta cell function-related genes of islets. Diabetes Res Clin Pract 77(Suppl 1):S150–S154

    Article  PubMed  CAS  Google Scholar 

  • Iozzo P, Beck-Nielsen H, Laakso M, Smith U, Yki-Jarvinen H, Ferrannini E (1999) Independent influence of age on basal insulin secretion in nondiabetic humans. European Group for the Study of Insulin Resistance. J Clin Endocrinol Metab 84:863–868

    Article  PubMed  CAS  Google Scholar 

  • Itakura E, Kishi C, Inoue K, Mizushima N (2008) Beclin 1 forms two distinct phosphatidylinositol 3-kinase complexes with mammalian Atg14 and UVRAG. Mol Biol Cell 19:5360–5372

    Article  PubMed  CAS  Google Scholar 

  • Juhasz G, Erdi B, Sass M, Neufeld TP (2007) Atg7-dependent autophagy promotes neuronal health, stress tolerance, and longevity but is dispensable for metamorphosis in Drosophila. Genes Dev 21:3061–3066

    Article  PubMed  CAS  Google Scholar 

  • Jung HS, Chung KW, Won Kim J, Kim J, Komatsu M, Tanaka K, Nguyen YH, Kang TM, Yoon KH, Kim JW et al (2008) Loss of autophagy diminishes pancreatic beta cell mass and function with resultant hyperglycemia. Cell Metab 8:318–324

    Article  PubMed  CAS  Google Scholar 

  • Kabeya Y, Mizushima N, Ueno T, Yamamoto A, Kirisako T, Noda T, Kominami E, Ohsumi Y, Yoshimori T (2000) LC3, a mammalian homologue of yeast Apg8p, is localized in autophagosome membranes after processing. EMBO J 19:5720–5728

    Article  PubMed  CAS  Google Scholar 

  • Kadowaki M, Karim MR (2009) Cytosolic LC3 ratio as a quantitative index of macroautophagy. Methods Enzymol 452:199–213

    Article  PubMed  CAS  Google Scholar 

  • Kang R, Zeh HJ, Lotze MT, Tang D (2011) The Beclin 1 network regulates autophagy and apoptosis. Cell Death Differ 18:571–580

    Article  PubMed  CAS  Google Scholar 

  • Kaniuk NA, Kiraly M, Bates H, Vranic M, Volchuk A, Brumell JH (2007) Ubiquitinated-protein aggregates form in pancreatic beta-cells during diabetes-induced oxidative stress and are regulated by autophagy. Diabetes 56:930–939

    Article  PubMed  CAS  Google Scholar 

  • Kinasiewicz A, Juszczak M, Pachecka J, Fiedor P (2004) Pancreatic islets isolation using different protocols with in situ flushing and intraductal collagenase injection. Physiol Res 53:327–333

    PubMed  CAS  Google Scholar 

  • Komatsu M, Waguri S, Chiba T, Murata S, Iwata J, Tanida I, Ueno T, Koike M, Uchiyama Y, Kominami E et al (2006) Loss of autophagy in the central nervous system causes neurodegeneration in mice. Nature 441:880–884

    Article  PubMed  CAS  Google Scholar 

  • Krishnamurthy J, Torrice C, Ramsey MR, Kovalev GI, Al-Regaiey K, Su L, Sharpless NE (2004) Ink4a/Arf expression is a biomarker of aging. J Clin Invest 114:1299–1307

    PubMed  CAS  Google Scholar 

  • Kurz T, Terman A, Brunk UT (2007) Autophagy, ageing and apoptosis: the role of oxidative stress and lysosomal iron. Arch Biochem Biophys 462:220–230

    Article  PubMed  CAS  Google Scholar 

  • Laybutt DR, Preston AM, Akerfeldt MC, Kench JG, Busch AK, Biankin AV, Biden TJ (2007) Endoplasmic reticulum stress contributes to beta cell apoptosis in type 2 diabetes. Diabetologia 50:752–763

    Article  PubMed  CAS  Google Scholar 

  • Lenaz G (2001) The mitochondrial production of reactive oxygen species: mechanisms and implications in human pathology. IUBMB Life 52:159–164

    Article  PubMed  CAS  Google Scholar 

  • Levine B, Kroemer G (2008) Autophagy in the pathogenesis of disease. Cell 132:27–42

    Article  PubMed  CAS  Google Scholar 

  • Maedler K, Schumann DM, Schulthess F, Oberholzer J, Bosco D, Berney T, Donath MY (2006) Aging correlates with decreased beta-cell proliferative capacity and enhanced sensitivity to apoptosis: a potential role for Fas and pancreatic duodenal homeobox-1. Diabetes 55:2455–2462

    Article  PubMed  CAS  Google Scholar 

  • Maiuri MC, Criollo A, Kroemer G (2010) Crosstalk between apoptosis and autophagy within the Beclin 1 interactome. EMBO J 29:515–516

    Article  PubMed  CAS  Google Scholar 

  • Maiuri MC, Zalckvar E, Kimchi A, Kroemer G (2007) Self-eating and self-killing: crosstalk between autophagy and apoptosis. Nat Rev Mol Cell Biol 8:741–752

    Article  PubMed  CAS  Google Scholar 

  • Marino G, Lopez-Otin C (2008) Autophagy and aging: new lessons from progeroid mice. Autophagy 4:807–809

    PubMed  CAS  Google Scholar 

  • Marsh BJ, Soden C, Alarcon C, Wicksteed BL, Yaekura K, Costin AJ, Morgan GP, Rhodes CJ (2007) Regulated autophagy controls hormone content in secretory-deficient pancreatic endocrine beta-cells. Mol Endocrinol 21:2255–2269

    Article  PubMed  CAS  Google Scholar 

  • Masini M, Bugliani M, Lupi R, del Guerra S, Boggi U, Filipponi F, Marselli L, Masiello P, Marchetti P (2009a) Autophagy in human type 2 diabetes pancreatic beta cells. Diabetologia 52:1083–1086

    Article  PubMed  CAS  Google Scholar 

  • Masini M, Lupi R, Bugliani M, Boggi U, Filipponi F, Masiello P, Marchetti P (2009b) A role for autophagy in beta-cell life and death. Islets 1:157–159

    Article  PubMed  Google Scholar 

  • Newsholme P, Haber EP, Hirabara SM, Rebelato EL, Procopio J, Morgan D, Oliveira-Emilio HC, Carpinelli AR, Curi R (2007) Diabetes associated cell stress and dysfunction: role of mitochondrial and non-mitochondrial ROS production and activity. J Physiol 583:9–24

    Article  PubMed  CAS  Google Scholar 

  • Pattison JS, Osinska H, Robbins J (2011) Atg7 induces basal autophagy and rescues autophagic deficiency in CryABR120G cardiomyocytes. Circ Res 109:151–160

    Article  PubMed  CAS  Google Scholar 

  • Perfetti R, Rafizadeh CM, Liotta AS, Egan JM (1995) Age-dependent reduction in insulin secretion and insulin mRNA in isolated islets from rats. Am J Physiol 269:E983–E990

    PubMed  CAS  Google Scholar 

  • Petersen KF, Dufour S, Befroy D, Garcia R, Shulman GI (2004) Impaired mitochondrial activity in the insulin-resistant offspring of patients with type 2 diabetes. N Engl J Med 350:664–671

    Article  PubMed  CAS  Google Scholar 

  • Rajawat YS, Bossis I (2008) Autophagy in aging and in neurodegenerative disorders. Hormones (Athens) 7:46–61

    Google Scholar 

  • Rajawat YS, Hilioti Z, Bossis I (2009) Aging: central role for autophagy and the lysosomal degradative system. Ageing Res Rev 8:199–213

    Article  PubMed  CAS  Google Scholar 

  • Rusten TE, Stenmark H (2010) p62, an autophagy hero or culprit? Nat Cell Biol 12:207–209

    Article  PubMed  CAS  Google Scholar 

  • Sakai K, Matsumoto K, Nishikawa T, Suefuji M, Nakamaru K, Hirashima Y, Kawashima J, Shirotani T, Ichinose K, Brownlee M et al (2003) Mitochondrial reactive oxygen species reduce insulin secretion by pancreatic beta-cells. Biochem Biophys Res Commun 300:216–222

    Article  PubMed  CAS  Google Scholar 

  • Sakuraba H, Mizukami H, Yagihashi N, Wada R, Hanyu C, Yagihashi S (2002) Reduced beta-cell mass and expression of oxidative stress-related DNA damage in the islet of Japanese Type II diabetic patients. Diabetologia 45:85–96

    Article  PubMed  CAS  Google Scholar 

  • Scheen AJ (2005) Diabetes mellitus in the elderly: insulin resistance and/or impaired insulin secretion? Diabetes Metab 31(2):5S27–25S34

    Article  PubMed  CAS  Google Scholar 

  • Seibenhener ML, Babu JR, Geetha T, Wong HC, Krishna NR, Wooten MW (2004) Sequestosome 1/p62 is a polyubiquitin chain binding protein involved in ubiquitin proteasome degradation. Mol Cell Biol 24:8055–8068

    Article  PubMed  CAS  Google Scholar 

  • Shigenaga MK, Gimeno CJ, Ames BN (1989) Urinary 8-hydroxy-2′-deoxyguanosine as a biological marker of in vivo oxidative DNA damage. Proc Natl Acad Sci U S A 86:9697–9701

    Article  PubMed  CAS  Google Scholar 

  • Sohal RS, Weindruch R (1996) Oxidative stress, caloric restriction, and aging. Science 273:59–63

    Article  PubMed  CAS  Google Scholar 

  • Toyokuni S, Tanaka T, Hattori Y, Nishiyama Y, Yoshida A, Uchida K, Hiai H, Ochi H, Osawa T (1997) Quantitative immunohistochemical determination of 8-hydroxy-2′-deoxyguanosine by a monoclonal antibody N45.1: its application to ferric nitrilotriacetate-induced renal carcinogenesis model. Lab Invest 76:365–374

    PubMed  CAS  Google Scholar 

  • Turrens JF (2003) Mitochondrial formation of reactive oxygen species. J Physiol 552:335–344

    Article  PubMed  CAS  Google Scholar 

  • Wohlgemuth SE, Julian D, Akin DE, Fried J, Toscano K, Leeuwenburgh C, Dunn WA Jr (2007) Autophagy in the heart and liver during normal aging and calorie restriction. Rejuvenation Res 10:281–292

    Article  PubMed  CAS  Google Scholar 

  • Wohlgemuth SE, Seo AY, Marzetti E, Lees HA, Leeuwenburgh C (2009) Skeletal muscle autophagy and apoptosis during aging: effects of calorie restriction and life-long exercise. Exp Gerontol 45:138–148

    Article  PubMed  Google Scholar 

  • Wohlgemuth SE, Seo AY, Marzetti E, Lees HA, Leeuwenburgh C (2010) Skeletal muscle autophagy and apoptosis during aging: effects of calorie restriction and life-long exercise. Exp Gerontol 45:138–148

    Article  PubMed  CAS  Google Scholar 

  • Wu JJ, Quijano C, Chen E, Liu H, Cao L, Fergusson MM, Rovira II, Gutkind S, Daniels MP, Komatsu M et al (2009) Mitochondrial dysfunction and oxidative stress mediate the physiological impairment induced by the disruption of autophagy. Aging (Albany NY) 1:425–437

    CAS  Google Scholar 

  • Yang W, Lu J, Weng J, Jia W, Ji L, **ao J, Shan Z, Liu J, Tian H, Ji Q et al (2010) Prevalence of diabetes among men and women in China. N Engl J Med 362:1090–1101

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

This work was supported by grants from the National Natural Science Foundation of China to C.L.L. (nos. 30873412, 81173625), to Y.L. (no. 30801201), from Bei**g Nova Program of China to Y.L. (2011116).

Author information

Authors and Affiliations

  1. Department of Geriatric Endocrinology, Chinese PLA General Hospital, 28 Fuxing Road, Bei**g, 100853, China

    Yu Liu, Zhaoyan Gu, Yingzhen Du, Minyan Liu, Shuangtong Yan, Jian Li, Yinghong Shao, Wenwen Zhong & Chunlin Li

  2. Department of Nephrology, State Key Lab of Kidney Diseases, Chinese PLA General Hospital, 28 Fuxing Road, Bei**g, 100853, China

    Suozhu Shi, Jianjun Gao & **angmei Chen

Authors
  1. Yu Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Suozhu Shi
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Zhaoyan Gu
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Yingzhen Du
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Minyan Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Shuangtong Yan
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Jianjun Gao
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Jian Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Yinghong Shao
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Wenwen Zhong
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. **angmei Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Chunlin Li
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to **angmei Chen or Chunlin Li.

Additional information

Yu Liu, Suozhu Shi and Zhaoyan Gu contributed equally to this work.

About this article

Cite this article

Liu, Y., Shi, S., Gu, Z. et al. Impaired autophagic function in rat islets with aging. AGE 35, 1531–1544 (2013). https://doi.org/10.1007/s11357-012-9456-0

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11357-012-9456-0

Keywords

Search

Navigation