Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

Global gene expression profile of normal and regenerating liver in young and old mice

Abstract

The ability of the liver to regenerate and adjust its size after two/third partial hepatectomy (PH) is impaired in old rodents and humans. Here, we investigated by microarray analysis the expression pattern of hepatic genes in young and old untreated mice and the differences in gene expression profile following PH. Of the 10,237 messenger RNAs that had detectable expression, only 108 displayed a greater than 2-fold modification in gene expression levels between the two groups. These genes were involved in inflammatory and immune response, xenobiotics, and lipid and glucose metabolism. To identify the genes responsible for the different regenerative response, 10-week and 18-month-old mice subjected to PH were sacrificed at different time intervals after surgery. The results showed that 2463 transcripts had significantly different expression post PH between the two groups. However, in spite of impaired liver regeneration in old mice, cell cycle genes were similarly modified in both groups, the only exception being cyclin D1 gene which was up-regulated soon after PH in young mice, but mostly down-regulated in aged animals. Surprisingly, while in young hepatectomized mice, Yap messenger RNA (mRNA) expression was not significantly enhanced and protein expression essentially reflected the progression into cell cycle, its mRNA and protein levels were robustly increased in the liver of aged animals. Furthermore, a significant change of the age-related expression of the size regulator Yes-associated protein (YAP) was observed. Unexpectedly, while in young hepatectomized mice, Yap mRNA expression was not significantly enhanced and protein expression essentially reflected the progression into cell cycle, its mRNA and protein levels were robustly increased in the liver of aged animals. Moreover, when PH was performed on mitogen-induced enlarged livers, the earlier restoration of the original liver mass compared to animals subjected to PH only led to YAP down-regulation concomitantly with cyclin D1 up-regulation. Our data suggest that YAP activation is a size-dependent homeostatic mechanism that does not necessarily reflect cell cycle progression.

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

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Abbreviations

AP-1:

Activating protein-1

BrdU:

Bromodeoxyuridine

Cebp:

CCAAT/enhancer-binding protein (C/EBP)

egr 1:

Early growth response-1

Foxm1:

Forkhead box M1

HGF:

Hepatocyte growth factor

Keap1:

Kelch-like ECH-associated protein 1

IL-6:

Interleukin-6

IHC:

Immunohistochemistry

NF-κB:

Nuclear factor-κB

Nqo2:

NAD(P)H dehydrogenase quinone 2

NRF2:

Nuclear factor (erythroid-derived 2)-like 2

PCNA:

Proliferating cell nuclear antigen

PH:

Partial hepatectomy

qRT-PCR:

Quantitative reverse transcriptase polymerase chain reaction

STAT3:

Signal transducer and activator of transcription factor 3

TAZ:

Transcriptional co-activator

TCPOBOP:

1,4-Bis[2-(3,5-dichloropyridyloxy)]benzene

TGF-β:

Transforming growth factor-β

TNF-α:

Tumor necrosis factor-α

YAP:

Yes-associated protein

References

  1. Ballou SP, Lozanski FB, Hodder S, Rzewnicki DL, Mion LC, Sipe JD, Ford AB, Kushner I (1996) Quantitative and qualitative alterations of acute-phase proteins in healthy elderly persons. Age Ageing 25:224–230

  2. Beyer TA, Xu W, Teupser D, auf dem Keller U, Bugnon P, Hildt E, Thiery J, Kan YW, Werner S (2008) Impaired liver regeneration in Nrf2 knockout mice: role of ROS-mediated insulin/IGF-1 resistance. EMBO J 27:212–223

  3. Bucher NLR, Swaffield MN, Di Troia JF (1964) The influence of age upon the incorporation of thymidine-2-C14 into the DNA of regenerating rat liver. Cancer Res 24:509–512

  4. Burzynski SR (2003) Gene silencing a new theory of aging. Med Hypotheses 60:578–583

  5. Burzynski SR (2005) Aging: gene silencing or gene activation? Med Hypotheses 64:201–208

  6. Camargo FD, Gokhale S, Johnnidis JB, Fu D, Bell GW, Jaenisch R et al (2007) YAP1 increases organ size and expands undifferentiated progenitor cells. Curr Biol 17:2054–2060

  7. Columbano A, Simbula M, Pibiri M, Perra A, Pisanu A, Uccheddu A, Ledda-Columbano GM (2008) Potential utility of xenobiotic mitogens in the context of liver regeneration in the elderly and living-related transplantation. Lab Investig 88:408–415

  8. Cressman DE, Greenbaum LE, DeAngelis RA, Ciliberto G, Furth EE, Poli V, Taub R (1996) Liver failure and defective hepatocyte regeneration in interleukin-6-deficient mice. Science 274:1379–1383

  9. Dong J, Feldmann G, Huang J, Wu S, Zhang N, Comerford SA et al (2007) Elucidation of a universal size-control mechanism in Drosophila and mammals. Cell 130:1120–1133

  10. Fausto N, Laird AD, Webber EM (1995) Role of growth factors and cytokines in hepatic regeneration. Faseb J 9:1527–1536

  11. Franceschi C, Bonafe M, Valensin S, Olivieri F, De Luca M, Ottaviani E, De Benedictis G (2000) Inflamm-aging. An evolutionary perspective on immunosenescence. Ann N Y Acad Sci 908:244–254

  12. Fry MF, Silber J, Loeb LA et al (1984) Delayed and reduced cell replication and diminishing levels of DNA polymerases-α in regenerating liver of aging mice. J Cell Physiol 118:225–232

  13. Gan L, Chitturi S, Farrell GC (2011) Mechanisms and implications of age-related changes in the liver: nonalcoholic fatty liver disease in the elderly. Curr Gerontol Geriatr Res 831536

  14. Grana X, Reddy PE (1995) Cell cycle control in mammalian cells, role of cyclins, cyclin-dependent kinases (CDKs), growth suppressor genes and cyclin dependent kinase inhibitors (CKIs) 11:211–219

  15. Grijalva JL, Huizenga M, Mueller K, Rodriguez S, Brazzo J, Camargo F, Sadri-Vakili G, Vakili K (2014) Dynamic alterations in Hippo signaling pathway and YAP activation during liver regeneration. Am J Physiol Gastrointest Liver Physiol 307:G196–G204

  16. Higgins GM, Anderson RM (1931) Experimental pathology of the liver. Restoration of the liver of the white rat following partial removal. Arch Pathol 12:186–202

  17. Iakova P, Awad SS, Timchenko N (2003) Aging reduced proliferative capacities of liver by switching pathway of C/EBPα growth arrest. Cell 113:495–506

  18. Köhler UA, Kurinna S, Schwitter D, Marti A, Schäfer M, Hellerbrand C, Speicher T, Werner S (2014) Activated Nrf2 impairs liver regeneration in mice by activation of genes involved in cell-cycle control and apoptosis. Hepatology 60:670–678

  19. Ledda-Columbano GM, Pibiri M, Cossu C, Molotzu F, Locker J, Columbano A (2004) Aging does not reduce the hepatocyte proliferative response of mice to the primary mitogen TCPOBOP. Hepatology 40:981–988

  20. Michalopoulos GK, DeFrances MC (1997) Liver regeneration. Science 276:60–66

  21. Pan D (2010) The hippo signaling pathway in development and cancer. Dev Cell 19:491–505

  22. Praticò D (2002) Lipid peroxidation and the aging process. Sci Aging Knowl Environ (50)re5

  23. Premoli A, Paschetta E, Hvalryg M, Spandre M, Bo S, Durazzo M (2009) Characteristics of liver diseases in the elderly: a review. Minerva Gastroenterol Dietol 55:71–78

  24. Schmucker DL (2005) Age-related changes in liver structure and function: implications for disease? Exp Gerontol 40:650–659

  25. Sheedfar F, Di Biase S, Koonen D, Vinciguerra M (2013) Liver diseases and aging: friends or foes? Aging Cell 12:950–954

  26. Singh P, Goode T, Dean A, Awad SS, Darlington GJ (2011) Elevated interferon gamma signaling contributes to impaired regeneration in the aged liver. J Gerontol A Biol Sci Med Sci 66:944–956

  27. Taub R (1996) Transcriptional control of liver regeneration. Faseb J 10:413–427

  28. Timchenko NA (2009) Aging and liver regeneration. Trends Endocrinol Metab 20(4):171–176

  29. Vranckx R, Savu L, Lambert N, de Conchard GV, Grosse R, Mourey MS, Corman B (1995) Plasma proteins as biomarkers of the aging process. Am J Physiol 268:R536–R548

  30. Wang X, Quail E, Hung NJ, Tan Y, Ye H, Costa RH (2001) Increased levels of forkhead box M1B transcription factor in transgenic mouse hepatocytes prevent age related proliferation defects in regenerating liver. PNAS 20:11468–11473

  31. Wang C, Zhang L, He Q, Feng X, Zhu J, Xu Z, Wang X, Chen F, Li X, Dong J (2012) Differences in Yes-associated protein and mRNA levels in regenerating liver and hepatocellular carcinoma. Mol Med Rep 5:410–414

  32. Yamada Y, Kirillova I, Peschon JJ, Fausto N (1997) Initiation of liver growth by tumor necrosis factor: deficient liver regeneration in mice lacking type I tumor necrosis factor receptor. Proc Natl Acad Sci USA 94(4):1441–1446

  33. Zhu C, Ikemoto T, Utsunomiya T, Yamada S, Morine Y, Imura S, Arakawa Y, Takasu C, Ishikawa D, Shimada M (2014) Senescence-related genes possibly responsible for poor liver regeneration after hepatectomy in elderly patients. J Gastroenterol Hepatol 29:1102–1108

Download references

Acknowledgments

Work supported by Italian Ministry of Health (Grant Young Researcher GR-2011-02350476 to M.R. and GR-2011-02347781to G.N.), Italian Ministry for Education, University and Research (Grants PRIN 2010LC747T), National Research Council Flagship Project Interomics, University of Salerno (FARB 2014), and Fondazione Banco di Sardegna to A.P.; G.N. is supported by a ‘Mario e Valeria Rindi’ fellowship of the Italian Foundation for Cancer Research. M.A.K. is a fellow of the Associazione Italiana Ricerca sul Cancro.

Author information

Correspondence to Monica Pibiri or Maria Ravo.

Additional information

Monica Pibiri and Pia Sulas contributed equally to this work.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary Table 1

(XLS 37 kb)

Supplementary Table 2

(XLS 123 kb)

Supplementary Table 3

(XLS 646 kb)

Supplementary Table 4

(XLS 58 kb)

Supplementary Table 5

(XLS 43 kb)

ESM 6

(DOC 27 kb)

About this article

Verify currency and authenticity via CrossMark

Cite this article

Pibiri, M., Sulas, P., Leoni, V.P. et al. Global gene expression profile of normal and regenerating liver in young and old mice. AGE 37, 59 (2015). https://doi.org/10.1007/s11357-015-9796-7

Download citation

Keywords

  • YAP
  • Cyclin D1
  • Microarray
  • Liver