Journal of Molecular Histology

, Volume 49, Issue 4, pp 347–356 | Cite as

Gamma-enolase predicts lung damage in severe acute pancreatitis-induced acute lung injury

  • Lawrence Owusu
  • Caiming Xu
  • Hailong ChenEmail author
  • Geliang Liu
  • Guixin Zhang
  • Jinwen Zhang
  • Zhankai Tang
  • Zhongwei Sun
  • Xin Yi
Original Paper


Severe acute pancreatitis (SAP) associated acute lung injury (ALI) accounts for about 70% mortality of SAP patients. However, there are no precise biomarkers for the disease currently. Herein, we evaluated the potential of gamma-enolase (ENO2), against its universal isoform alpha-enolase (ENO1), as a marker of SAP–ALI in a rat model. Firstly, 16 male Sprague–Dawley rats were randomly divided into two groups, Sham (n = 8) and SAP–ALI (n = 8), for pancreatitis induction. Ultra-structure examination by electron microscopy and HE staining were used for lung injury assessment. Lung tissue expressions of alpha-enolase and gamma-enolase were evaluated by qRT-PCR and immunohistochemistry. In a prospective validation experiment, 28 rats were used: sham (n = 8), SAP–ALI at 3 h (3 h, n = 10), and SAP–ALI at 24 h (24 h, n = 10). Lung tissue damage, tissue expression and circulating alpha-enolase and gamma-enolase levels were evaluated. Elevated serum levels of α-amylase and TNF-α were observed in SAP rats but not in sham-operated rats. Histological examination of pancreatic and lung tissues indicated marked damage in SAP rats. While alpha-enolase was universally expressed, gamma-enolase was expressed only in damaged lung tissues. Gamma-enolase was detected in lung tissues, BALF, and serum as early as 3 h post-surgery when physical pathological damage was not apparent. Unlike alpha-enolase, secreted and/or circulating gamma-enolase level progressively increased, especially in serum, as lung damage progressed. Thus, gamma-enolase may signal and correlate lung tissue damage well before obvious physical pathological tissue damage and might be a candidate diagnostic and/or prognostic marker.


Severe acute pancreatitis-induced acute lung injury Alpha-enolase Gamma-enolase Sprague–Dawley rats 



Support for this study was by a grant from the National Natural Science Foundation of China (No. 81573751) to HC. However, the funding body had no influence on the study design, data interpretation, or manuscript preparation.

Author contributions

LO and HC conceived and designed the experiment; CX, GL, JZ, ZT, ZS and LO carried out the animal model and sample collections; LO, CX, ZT, JZ and ZS carried out validation experiments; HC secured funding for the study, and with LO drafted the manuscript; All authors critically reviewed and approved the final manuscript.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no competing interest concerning this publication.


  1. Abraham E, Carmody A, Shenkar R, Arcaroli J (2000) Neutrophils as early immunologic effectors in hemorrhage- or endotoxemia induced acute lung injury. Am J Physiol Lung Cell Mol Physiol 279:L1137–L1145CrossRefGoogle Scholar
  2. Acosta JM, Ledesma CL (1974) Gallstone migration as a cause of acute pancreatitis. N Engl J Med 290:484–487CrossRefPubMedGoogle Scholar
  3. Allen FJ, Van Velden DJ, Heyns CF (1995) Are neuroendocrine cells of practical value as an independent prognostic parameter in prostate cancer? Br J Urol 75:751–754CrossRefPubMedGoogle Scholar
  4. Bhargava M, Becker TL, Vilken KJ, Jagtap PD, Dey S, Steinbach MS, Wu B, Kumar V, Bitterman PB, Ingbar DH, Wendt CH (2014) Proteomic profile in acute respiratory distress syndrome differentiates survivors from non-survivors. PLoS ONE 9:e109713. CrossRefPubMedPubMedCentralGoogle Scholar
  5. Blondonnet R, Constantin J-M, Sapin V, Jabaudon M (2016) A pathophysiologic approach to biomarkers in acute respiratory distress syndrome. Dis Markers. PubMedPubMedCentralCrossRefGoogle Scholar
  6. Collazos J, Esteban C, Fernandez A, Genolla J (1994) Measurement of the serum tumor marker neuron-specific enolase in patients with benign pulmonary diseases. Am J Respir Crit Care Med 150:143–145CrossRefPubMedGoogle Scholar
  7. Dervenis C, Johnson CD, Bassi C, Bradley E, Imrie CW, McMahon MJ, Modlin I (1999) Diagnosis, objective assessment of severity, and management of acute pancreatitis: Santorini consensus conference. Int J Pancreatol 25:195–210PubMedGoogle Scholar
  8. Díaz-Ramos A, Roig-Borrellas A, García-Melero A, López-Alemany R (2012) α-Enolase, a multifunctional protein: its role on pathophysiological situations. J Biomed Biotechnol. PubMedPubMedCentralCrossRefGoogle Scholar
  9. Fischer AH, Jacobson KA, Rose J, Zeller R (2008) Hematoxylin and eosin staining of tissue and cell sections. Cold Spring Harb Protoc. CrossRefGoogle Scholar
  10. Flexner S (1906) The constituents of the bile causing pancreatitis and the effect of colloids upon its action. J Exp Med 8:167–177CrossRefPubMedPubMedCentralGoogle Scholar
  11. Fremont RD, Koyama T, Calfee CS, Wu W, Dossett LA, Bossert FR, Mitchell D, Wickersham N, Bernard GR, Matthay MA, May AK, Ware LB (2010) Acute lung injury in patients with traumatic injuries: utility of a panel of biomarkers for diagnosis and pathogenesis. J Trauma 68:1121–1127CrossRefPubMedPubMedCentralGoogle Scholar
  12. Guice KS, Oldham KT, Johnson KJ, Kunkel RG, Morganroth ML, Ward PA (1988) Pancreatitis-induced acute lung injury: an ARDS model. Ann Surg 208:71–77CrossRefPubMedPubMedCentralGoogle Scholar
  13. Haimoto H, Takahashi Y, Koshikawa T, Nagura H, Kato K (1985) Immunohistochemical localization of gamma-enolase in normal human tissues other than nervous and neuroendocrine tissues. Lab Invest 52:257–263PubMedGoogle Scholar
  14. Hao X, Sun B, Hu L et al (2004) Differential gene and protein expression in primary breast malignancies and their lymph node metastases as revealed by combined cDNA microarray and tissue microarray analysis. Cancer 100:1110–1122CrossRefPubMedGoogle Scholar
  15. Isgrò MA, Bottoni P, Scatena R (2015) Neuron-specific enolase as a biomarker: biochemical and clinical aspects. Adv Exp Med Biol 867:125–143CrossRefPubMedGoogle Scholar
  16. Kasprzak A, Zabel M, Biczysko W (2007) Selected markers (chromogranin A, neuron-specific enolase, synaptophysin, protein gene product 9.5) in diagnosis and prognosis of neuroendocrine pulmonary tumours. Pol J Pathol 58:23–33PubMedGoogle Scholar
  17. Kondoh H, Lleonart ME, Bernard D, Gil J (2007) Protection from oxidative stress by enhanced glycolysis; a possible mechanism of cellular immortalization. Histol Histopathol 22:85–90PubMedGoogle Scholar
  18. Konrad FM, Reutershan J (2012) CXCR2 in acute lung injury. Mediat Inflamm. CrossRefGoogle Scholar
  19. Lamerz R (1998) NSE (neuron-specific enolase) γ-enolase. In: Thomas L (ed) Clinical laboratory diagnostics: use and assessment of clinical laboratory results, 1st edn. TH-Books Verlagsgesellschaft, Frankfurt, pp 979–981Google Scholar
  20. Leiherer A, Stoemmer K, Muendlein A, Saely CH, Kinz E, Brandtner EM, Fraunberger P, Drexel H (2016) Quercetin impacts expression of metabolism and obesity-associated genes in SGBS adipocytes. Nutrients 8:E282. CrossRefPubMedGoogle Scholar
  21. Liu G, Zhang J, Chen H, Wang C, Qiu Y, Liu Y, Wan J, Guo H (2014) Effects and mechanisms of alveolar type II epithelial cell apoptosis in severe pancreatitis-induced acute lung injury. Exp Ther Med 7:565–572CrossRefPubMedGoogle Scholar
  22. Luan Z-G, Zhang J, Yin X-H, Ma X-C, Guo R-X (2013) Ethyl pyruvate significantly inhibits tumour necrosis factor-a, interleukin-1b and high mobility group box 1 releasing and attenuates sodium taurocholate-induced severe acute pancreatitis associated with acute lung injury. Clin Exp Immunol 172:417–426CrossRefPubMedPubMedCentralGoogle Scholar
  23. Mair J (1997) Progress in myocardial damage detection: new biochemical markers for clinicians. Crit Rev Clin Lab Sci 34:1–66CrossRefPubMedGoogle Scholar
  24. Nakatsuka S, Nishiu M, Tomita Y, Miwa H, Takakuwa T, Iuchl K, Yamamoto S, Aozasa K (2002) Enhanced expression of neuron-specific enolase (NSE) in pyothorax-associated lymphoma (PAL). Jpn J Cancer Res 93:411–416CrossRefPubMedPubMedCentralGoogle Scholar
  25. Niklinski J, Furman M (1995) Clinical tumour markers in lung cancer. Eur J Cancer Prev 4:129–138CrossRefPubMedGoogle Scholar
  26. Renzulli P, Jakob SM, Täuber M, Candinas D, Gloor B (2005) Severe acute pancreatitis: case-oriented discussion of interdisciplinary management. Pancreatology 5:145–156CrossRefPubMedGoogle Scholar
  27. Rubenfeld GD, Caldwell E, Peabody E, Weaver J, Martin DP, Neff M, Stern EJ, Hudson LD (2005) Incidence and outcomes of acute lung injury. N Engl J Med 353:1685–1693CrossRefPubMedGoogle Scholar
  28. Samuel I, Yuan Z, Meyerholz DK, Twait E, Willard DE, Kempurai D (2010) A novel model of severe gallstone pancreatitis: murine pancreatic duct ligation results in systemic inflammation and substantial mortality. Pancreatology 10:536–544CrossRefPubMedPubMedCentralGoogle Scholar
  29. Schmechel D, Marangos PJ, Brightman M (1978) Neuron-specific enolase is a molecular marker for peripheral and central neuroendocrine cells. Nature 276:834–836CrossRefPubMedGoogle Scholar
  30. Soh MA, Garrett SH, Somji S, Dunlevy JR, Zhou XD, Sens MA, Bethula CS, Allen C, Sens DA (2011) Arsenic, cadmium and neuron specific enolase (gamma-enolase, γ-enolase) expression in breast cancer. Cancer Cell Int 11:41. CrossRefPubMedPubMedCentralGoogle Scholar
  31. Stelzl T, von Bose MJ, Hogl B, Fuchs HH, Flugel KA (1995) A comparison of the prognostic value of neuron-specific enolase serum levels and somatosensory evoked potentials in 13 reanimated patients. Eur J Emerg Med 2:24–27CrossRefPubMedGoogle Scholar
  32. Suresh MR (2005) Cancer markers. In: Wild D (ed) The immunoassay handbook, 3rd edn. Elsevier, Oxford, pp 664–694Google Scholar
  33. Takashima M, Kuramitsu Y, Yokoyama Y, Iizuka N, Fujimoto M, Nishisaka T, Okita K, Oka M, Nakamura K (2005) Overexpression of alpha enolase in hepatitis C virus-related hepatocellular carcinoma: association with tumor progression as determined by proteomic analysis. Proteomics 5:1686–1692CrossRefPubMedGoogle Scholar
  34. Tang M, Tian Y, Li D, Lv J, Li Q, Kuang C, Hu P, Wang Y, Wang J, Su K, Wei L (2014) TNF-α mediated increase of HIF-1α inhibits VASP expression, which reduces alveolar-capillary barrier function during acute lung injury (ALI). PLoS ONE 9:e102967CrossRefPubMedPubMedCentralGoogle Scholar
  35. Tiainen M, Roine RO, Pettila V, Takkunen O (2003) Serum neuron-specific enolase and S-100B protein in cardiac arrest patients treated with hypothermia. Stroke 34:2881–2886CrossRefPubMedGoogle Scholar
  36. Vizin T, Kos J (2015) Gamma-enolase: a well-known tumour marker, with a less-known role in cancer. Radiol Oncol 49:217–226CrossRefPubMedPubMedCentralGoogle Scholar
  37. Xu H, Ye X, Steinberg H, Liu SF (2010) Selective blockade of endothelial NF-kappaB pathway differentially affects systemic inflammation and multiple organ dysfunction and injury in septic mice. J Pathol 220:490–498PubMedGoogle Scholar
  38. Yan T, Skaftnesmo KO, Leiss L, Sleire L, Wang J, Li X, Enger P (2011) Neuronal markers are expressed in human gliomas and NSE knockdown sensitizes glioblastoma cells to radiotherapy and temozolomide. BMC Cancer 11:524. CrossRefPubMedPubMedCentralGoogle Scholar
  39. Zhang JW, Zhang GX, Chen HL, Liu GL, Owusu L, Wang YX, Wang GY, Xu CM (2015) Therapeutic effect of Qingyi decoction in severe acute pancreatitis-induced intestinal barrier injury. World J Gastroenterol 21:3537–3546CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer Science+Business Media B.V., part of Springer Nature 2018

Authors and Affiliations

  1. 1.Department of Surgery, The First Affiliated HospitalDalian Medical UniversityDalianChina
  2. 2.Department of Integrative MedicineDalian Medical UniversityDalianChina
  3. 3.Department of Surgery, The Second Affiliated HospitalDalian Medical UniversityDalianChina
  4. 4.Department of Traditional Chinese MedicineDalian Obstetrics and Gynecology HospitalDalianChina
  5. 5.Department of Biochemistry and BiotechnologyKwame Nkrumah University of Science and Technology (KNUST), PMB, UPOKumasiGhana
  6. 6.Department of BiotechnologyDalian Medical UniversityDalianChina

Personalised recommendations