Journal of Physiology and Biochemistry

, Volume 74, Issue 1, pp 25–33 | Cite as

The differential expression of novel circular RNAs in an acute lung injury rat model caused by smoke inhalation

  • Zhiqiang Ye
  • Xuhui Liu
  • Yuewu Yang
  • Xianling Zhang
  • Ting Yu
  • Shigeng Li
  • Yawei Feng
  • Gangjian LuoEmail author
Original Article


Acute lung injury caused by smoke inhalation is a common severe clinical syndrome. This study aimed to investigate the potential expression of circular RNAs during acute lung injury triggered by smoke inhalation. The acute lung injury rat model was established with smoke inhalation from a self-made smoke generator. The occurrence of acute lung injury was validated by an analysis of the bronchoalveolar lavage fluid and hematoxylin-eosin (HE) staining of lung tissues. Next-generation sequencing and quantitative PCR were performed to identify the differentially expressed circular RNAs associated with acute lung injury that was caused by smoke inhalation. The circular form of the identified RNAs was finally verified by multiple RT-PCR-based assays. The bronchoalveolar lavage fluid (BALF) and lung tissue analysis showed that smoke inhalation successfully induced acute injury in rats, as evidenced by the significantly altered cell numbers, including macrophages, neutrophils, and red blood cells, disrupted cell lining, and increased levels of interleukin-1β, tumor necrosis factor-alpha, and IL-8 in lung tissues. Ten significantly differentially expressed circular RNAs were identified with next-generation sequencing and RT-PCR. The circular form of these RNAs was verified by multiple RT-PCR-based assays. In conclusion, the identified circular RNAs were prevalently and differentially expressed in rat lungs after acute lung injury caused by smoke inhalation.


Acute lung injury Smoke inhalation Circular RNA Rat model 



This study was supported by Science and Technology Plan Projects of Guangdong Province (Nos. 2014A020212533 and 2014A020212707).

Compliance with ethical standards

All experiments were authorized by the Animal Care and Ethics Committee of Sun Yat-sen University.

Conflict of interest

The authors have no conflicts of interest to declare.


  1. 1.
    Abdelmohsen K, Panda AC, De S, Grammatikakis I, Kim J, Ding J, Ji HN, Kim KM, Mattison JA, Cabo RD (2015) Circular RNAs in monkey muscle: age-dependent changes. Aging 7(11):903–910.  10.18632/aging.100834 CrossRefPubMedPubMedCentralGoogle Scholar
  2. 2.
    Chen LL (2016) The biogenesis and emerging roles of circular RNAs. Nat Rev Mol Cell Biol 17(4):205–211. CrossRefPubMedGoogle Scholar
  3. 3.
    Chen LL, Yang L (2015) Regulation of circRNA biogenesis. RNA Biol 12(4):381–388. CrossRefPubMedPubMedCentralGoogle Scholar
  4. 4.
    Cheng J, Metge F, Dieterich C (2016) Specific identification and quantification of circular RNAs from sequencing data. Bioinformatics 32(7):1094–1096. CrossRefPubMedGoogle Scholar
  5. 5.
    Enkhbaatar P, Traber DL (2004) Pathophysiology of acute lung injury in combined burn and smoke inhalation injury. Clin Sci 107(2):137–143. CrossRefPubMedGoogle Scholar
  6. 6.
    Greene J, Baird AM, Brady L, Lim M, Gray SG, Mcdermott R & Finn SP. (2017). Circular RNAs: biogenesis, function and role in human diseases. Front Mol Biosci. 4:38.
  7. 7.
    Guo Z, Wen Z, Qin A, Zhou Y, Liao Z, Liu Z, Liang Y, Ren T, Xu L (2013) Antisense oligonucleotide treatment enhances the recovery of acute lung injury through IL-10ق€ secreting M2-like macrophage-induced expansion of CD4+ regulatory T cells. J Immunol 190(8):4337–4348. CrossRefPubMedPubMedCentralGoogle Scholar
  8. 8.
    Hansen TB, Jensen TI, Clausen BH, Bramsen JB, Finsen B, Damgaard CK, Kjems J (2013) Natural RNA circles function as efficient microRNA sponges. Nature 495(7441):384–388. CrossRefPubMedGoogle Scholar
  9. 9.
    Johnson ER, Matthay MA (2010) Acute lung injury: epidemiology, pathogenesis, and treatment. J Aerosol Med Pulmo Drug Delivery 23:243CrossRefGoogle Scholar
  10. 10.
    Khimenko PL, Taylor AE (1999) Segmental microvascular permeability in ischemia-reperfusion injury in rat lung. Am J Physiol 276(6 Pt 1):L958–L960PubMedGoogle Scholar
  11. 11.
    Ku CM, Lin JY (2015) Farnesol, a sesquiterpene alcohol in herbal plants, exerts anti-inflammatory and antiallergic effects on ovalbumin-sensitized and -challenged asthmatic mice. Ev-Based Complementary Alternat Med 2015(2015–4-19):1–12CrossRefGoogle Scholar
  12. 12.
    Matute-Bello G, Frevert CW, Martin TR (2008) Animal models of acute lung injury. Am J Physiol Lung Cell Mol Physiol 295:379–399CrossRefGoogle Scholar
  13. 13.
    Petar G, Panagitis P, Nikolaus R (2014) circBase: a database for circular RNAs. Rna-a Publ Rna Soc 20:1666CrossRefGoogle Scholar
  14. 14.
    Pei W, Tao L, Zhang LW, Zhang S, Cao J, Jiao Y, Tong J, Nie J (2017) Circular RNA profiles in mouse lung tissue induced by radon. Environ Health Prev Med 22:36CrossRefPubMedPubMedCentralGoogle Scholar
  15. 15.
    Sanger HL, Klotz G, Riesner D, Gross HJ, Kleinschmidt AK (1976) Viroids are single-stranded covalently closed circular RNA molecules existing as highly base-paired rod-like structures. Proc Natl Acad Sci U S A 73(11):3852–3856. CrossRefPubMedPubMedCentralGoogle Scholar
  16. 16.
    Soejima K, Schmalstieg FC, Sakurai H, Traber LD, Traber DL (2001) Pathophysiological analysis of combined burn and smoke inhalation injuries in sheep. Am J Physiol Lung Cell Mol Physiol 280(6):L1233–L1241CrossRefPubMedGoogle Scholar
  17. 17.
    Subbiah R, Pattarayan D, R P PSG, Thimmulappa RK (2016) MicroRNA regulation of acute lung injury and acute respiratory distress syndrome. J Cell Physiol 231(10):2097–106CrossRefGoogle Scholar
  18. 18.
    Tsushima K, King LN (2009) Acute lung injury review. Intern Med 48(9):621–630. CrossRefPubMedGoogle Scholar
  19. 19.
    Wan L, Zhang L, Fan K, Cheng ZX, Sun QC, Wang JJ Circular RNA-ITCH suppresses lung cancer proliferation via inhibiting the Wnt/خ -catenin pathway. Biomed Res Int 2016, 2016:1–11Google Scholar
  20. 20.
    Wheeler AP, Bernard GR (2007) Acute lung injury and the acute respiratory distress syndrome: a clinical review. Lancet 369(9572):1553–1564. CrossRefPubMedGoogle Scholar
  21. 21.
    Xu Z, Zhang C, Cheng L, Hu M, Tao H, Song L (2014) The microRNA miR-17 regulates lung FoxA1 expression during lipopolysaccharide-induced acute lung injury. Biochem Biophys Res Communications 445:48–53CrossRefGoogle Scholar
  22. 22.
    You X, Conrad TO (2016) Acfs: accurate circRNA identification and quantification from RNA-Seq data. Sci Rep 6(1):38820. CrossRefPubMedPubMedCentralGoogle Scholar
  23. 23.
    Zhang P, Zuo Z, Shang W, Wu A, Bi R, Wu J, Li S, Sun X, Jiang L (2017) Identification of differentially expressed circular RNAs in human colorectal cancer. Tumour Biol J Int Soc Oncodev Biol Med 39:1010428317694546Google Scholar
  24. 24.
    Zhou T, Garcia JGN, Zhang W (2011) Integrating microRNAs into a system biology approach to acute lung injury. Transl Res J Lab Clin Med 157:180–190CrossRefGoogle Scholar
  25. 25.
    Zhu X, Wang X, Wei S, Chen Y, Chen Y, Fan X, Han S & Wu G. (2017). hsa_circ_0013958: a circular RNA and potential novel biomarker for lung adenocarcinoma. FEBS J 284(14):2170–2182Google Scholar

Copyright information

© University of Navarra 2017

Authors and Affiliations

  • Zhiqiang Ye
    • 1
  • Xuhui Liu
    • 1
  • Yuewu Yang
    • 2
  • Xianling Zhang
    • 3
  • Ting Yu
    • 1
  • Shigeng Li
    • 1
  • Yawei Feng
    • 4
  • Gangjian Luo
    • 4
    Email author
  1. 1.Department of Emergency, The Third Affiliated HospitalSun Yat-sen UniversityGuangzhouChina
  2. 2.Department of Traditional Chinese Medicine, The Third Affiliated HospitalSun Yat-sen UniversityGuangzhouChina
  3. 3.Department of Hepatic Surgery, The Third Affiliated HospitalSun Yat-sen UniversityGuangzhouChina
  4. 4.Department of Anesthesiology, The Third Affiliated HospitalSun Yat-sen UniversityGuangzhouChina

Personalised recommendations