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Human Cell

pp 1–10 | Cite as

Pathophysiological significance of clock genes BMAL1 and PER2 as erythropoietin-controlling factors in acute blood hemorrhage

  • Naoto TaniEmail author
  • Tomoya Ikeda
  • Yayoi Aoki
  • Alissa Shida
  • Shigeki Oritani
  • Takaki Ishikawa
Research Article
  • 7 Downloads

Abstract

This study aimed to characterize the pathophysiology, including possible correlations, of clock gene expression and erythropoietin (EPO) production in the acute stage of blood hemorrhage. Specimens of human cortical tissues (right and left kidneys) and cardiac blood were collected at autopsy from 52 cases following mortality due to acute-stage blood hemorrhage following sharp instrument injury. BMAL1 and PER2 mRNA levels were determined by reverse transcription-polymerase chain reaction; BMAL1 and PER2 protein levels were assessed using immunohistochemistry; BMAL1 protein levels were quantitatively measured by western blotting; and serum EPO levels were measured by chemiluminescent enzyme immunoassay. Separately, a rat model of hemorrhagic conditions was generated and used to confirm the results obtained with autopsy-derived specimens. A positive correlation was observed between BMAL1 protein and serum EPO levels, but not between BMAL1 mRNA levels and serum EPO levels. We also noted that Per2 mRNA expression became elevated in humans who survived for > 3 h after acute hemorrhagic events, with subsequent decreases in serum EPO levels. The rat model showed that even short (30-min) intervals of blood loss yielded increases in both Bmal1 mRNA and serum EPO levels; longer (60-min) intervals resulted in increases in Per2 mRNA expression along with decreases in serum EPO. Thus, the acute-stage human hemorrhage cases and the rat hemorrhage model yielded similar tendencies for clock gene expression and EPO secretion. In conclusion, our results indicated that clock genes are involved in the regulation of EPO production during the early stages of hypoxia/ischemia resulting from the acute hemorrhagic events.

Keywords

Forensic Clock gene Erythropoietin Hemorrhagic death/shock Hypoxia/ischemia 

Notes

Funding

The authors received no specific funding for this work.

Compliance with ethical standards

Conflict of interest

The authors declare they have no conflict of interest.

Statement of human rights

All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards. The independent Ethics Committee at Osaka City University Graduate School of Medicine approved this study (Authorization no. 2001).

Statement on the welfare of animals

All applicable international, national, and/or institutional guidelines for the care and use of animals were followed. All procedures performed in studies involving animals were in accordance with the ethical standards of the institution or practice at which the studies were conducted.

References

  1. 1.
    Cannon JW. Hemorrhagic shock. N Engl J Med. 2018;378:370–9.CrossRefPubMedGoogle Scholar
  2. 2.
    Pilorz V, Helfrich-Förster C, Oster H. The role of the circadian clock system in physiology. Pflugers Arch. 2018;470:227–39.CrossRefPubMedGoogle Scholar
  3. 3.
    Gumz ML. Molecular basis of circadian rhythmicity in renal physiology and pathophysiology. Exp Physiol. 2016;101:1025–9.CrossRefPubMedGoogle Scholar
  4. 4.
    Stow LR, Gumz ML. The circadian clock in the kidney. J Am Soc Nephrol. 2011;22:598–604.CrossRefPubMedGoogle Scholar
  5. 5.
    Solocinski K, Gumz ML. The circadian clock in the regulation of renal rhythms. J Biol Rhythms. 2015;30:470–86.CrossRefPubMedGoogle Scholar
  6. 6.
    Quan L, Zhu BL, Ishikawa T, et al. Postmortem serum erythropoietin level as a marker of survival time in injury deaths. Forensic Sci Int. 2010;200:117–22.CrossRefPubMedGoogle Scholar
  7. 7.
    Firsov D, Bonny O. Circadian regulation of renal function. Kidney Int. 2010;78:640–5.CrossRefPubMedGoogle Scholar
  8. 8.
    Yoshimura A, Arai K. Physician education: the erythropoietin receptor and signal transduction. Oncologist. 1996;1:337–9.PubMedGoogle Scholar
  9. 9.
    Wu Y, Tang D, Liu N, et al. Reciprocal regulation between the circadian clock and hypoxia signaling at the genome level in mammals. Cell Metab. 2017;25:73–85.CrossRefPubMedGoogle Scholar
  10. 10.
    Sousa Fialho MDL, Abd Jamil AH, Stannard GA, Heather LC. Hypoxia-inducible factor 1 signalling, metabolism and its therapeutic potential in cardiovascular disease. Biochim Biophys Acta Mol Basis Dis. 2019;1865:831–43.CrossRefPubMedGoogle Scholar
  11. 11.
    Benderro GF, LaManna JC. Kidney EPO expression during chronic hypoxia in aged mice. Adv Exp Med Biol. 2013;765:9–14.CrossRefPubMedGoogle Scholar
  12. 12.
    Suzuki N, Obara N, Yamamoto M. Use of gene-manipulated mice in the study of erythropoietin gene expression. Methods Enzymol. 2007;435:157–77.CrossRefPubMedGoogle Scholar
  13. 13.
    Morita M, Ohneda O, Yamashita T, Takahashi S, Suzuki N, Nakajima O, Kawauchi S, Ema M, Shibahara S, Udono T, Tomita K, Tamai M, Sogawa K, Yamamoto M, Fujii-Kuriyama Y. HLF/HIF-2alpha is a key factor in retinopathy of prematurity in association with erythropoietin. EMBO J. 2003;22:1134–46.CrossRefPubMedGoogle Scholar
  14. 14.
    Ranjbaran M, Kadkhodaee M, Seifi B. Renal tissue pro-inflammatory gene expression is reduced by erythropoietin in rats subjected to hemorrhagic shock. J Nephropathol. 2017;6:69–73.CrossRefPubMedGoogle Scholar
  15. 15.
    Tani N, Ikeda T, Oritani S, Michiue T, Ishikawa T. Role of circadian clock genes in sudden cardiac death: a pilot study. J Hard Tissue Biol. 2017;26:347–54.CrossRefGoogle Scholar
  16. 16.
    Wang Q, Ishikawa T, Michiue T, Zhu BL, Guan DW, Maeda H. Stability of endogenous reference genes in postmortem human brains for normalization of quantitative real-time PCR data: comprehensive evaluation using geNorm, NormFinder, and BestKeeper. Int J Legal Med. 2012;126:943–52.CrossRefPubMedGoogle Scholar
  17. 17.
    Lin R, Mo Y, Zha H, et al. CLOCK acetylates ASS1 to drive circadian rhythm of ureagenesis. Mol Cell. 2017;68:198–209.e6.CrossRefPubMedGoogle Scholar
  18. 18.
    Lee Y, Jang AR, Francey LJ, Sehgal A, Hogenesch JB. KPNB1 mediates PER/CRY nuclear translocation and circadian clock function. Elife. 2015; 4.Google Scholar
  19. 19.
    Kawamoto O, Michiue T, Ishikawa T, Maeda H. Immunohistochemistry of connexin43 and zonula occludens-1 in the myocardium as markers of early ischemia in autopsy material. Histol Histopathol. 2014;29:767–75.PubMedGoogle Scholar
  20. 20.
    Sato H, Tanaka T, Tanaka N. The effect of p38 mitogen-activated protein kinase activation on inflammatory liver damage following hemorrhagic shock in rats. PLoS One. 2012;7:e30124.CrossRefPubMedGoogle Scholar
  21. 21.
    Zhao D, Michiue T, Maeda H. Tissue-dependent VEGF and GLUT1 induction in a rat hemorrhage model: With regard to diagnostic application of mRNA quantification in forensic pathology. Forensic Sci Int. 2015;255:118–22.CrossRefPubMedGoogle Scholar
  22. 22.
    Vara-Gama N, Valladares-Méndez A, Navarrete-Vazquez G, Estrada-Soto S, Orozco-Castellanos LM, Rivera-Leyva JC. Biopharmaceutical characterization and bioavailability study of a tetrazole analog of clofibric acid in rat. Molecules. 2017;22:282.CrossRefGoogle Scholar
  23. 23.
    Yoshida C, Ishikawa T, Michiue T, Quan L, Maeda H. Postmortem biochemistry and immunohistochemistry of chromogranin A as a stress marker with special regard to fatal hypothermia and hyperthermia. Int J Legal Med. 2011;125:11–20.CrossRefPubMedGoogle Scholar
  24. 24.
    Zhu BL, Ishida K, Quan L, et al. Postmortem serum uric acid and creatinine levels in relation to the causes of death. Forensic Sci Int. 2002;125:59–66.CrossRefPubMedGoogle Scholar
  25. 25.
    Bozek K, Relógio A, Kielbasa SM, et al. Regulation of clock-controlled genes in mammals. PLoS One. 2009;4:e4882.CrossRefPubMedGoogle Scholar
  26. 26.
    Mazzoccoli G, De Cata A, Piepoli A, Vinciguerra M. The circadian clock and the hypoxic response pathway in kidney cancer. Tumour Biol. 2014;35:1–7.CrossRefPubMedGoogle Scholar
  27. 27.
    Lemmer B, Oster H. The role of circadian rhythms in the hypertension of diabetes mellitus and the metabolic syndrome. Curr Hypertens Rep. 2018;20:43.CrossRefPubMedGoogle Scholar
  28. 28.
    Sun B, Feng X, Ding X, et al. Expression of Clock genes in the pineal glands of newborn rats with hypoxic-ischemic encephalopathy. Neural Regen Res. 2012;7:2221–6.PubMedGoogle Scholar
  29. 29.
    Curtis AM, Bellet MM, Sassone-Corsi P, O’Neill LA. Circadian clock proteins and immunity. Immunity. 2014;40:178–86.CrossRefPubMedGoogle Scholar
  30. 30.
    Bonney S, Kominsky D, Brodsky K, Eltzschig H, Walker L, Eckle T. Cardiac Per2 functions as novel link between fatty acid metabolism and myocardial inflammation during ischemia and reperfusion injury of the heart. PLoS One. 2013;8:e71493.CrossRefPubMedGoogle Scholar
  31. 31.
    Panjeta M, Tahirović I, Sofić E, Ćorić J, Dervišević A. Interpretation of erythropoietin and haemoglobin levels in patients with various stages of chronic kidney disease. J Med Biochem. 2017;36:145–52.CrossRefPubMedGoogle Scholar

Copyright information

© Japan Human Cell Society and Springer Japan KK, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of Legal MedicineOsaka City University Medical SchoolAbenoJapan
  2. 2.Forensic Autopsy SectionMedico-Legal Consultation and Postmortem Investigation Support Center (MLCPI-SC)OsakaJapan

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