Characteristics of Changes in Inflammatory Cytokines as a Function of Hepatic Ischemia-Reperfusion Injury Stage in Mice

  • Shi-peng LiEmail author
  • Fei-fei Wang
  • Wen-kui Zhang
  • Ming-ze Bian
  • Shen-yan Zhang
  • Han Yan
  • Yuan Fang
  • Hai-ming ZhangEmail author
Original Article


Liver ischemia-reperfusion injury (IRI) can severely compromise the prognosis of patients receiving liver surgery. While inflammation contributes to the damage resulting from IRI, only a limited number of inflammation biomarkers have been identified as being associated with the different stages of hepatic IRI. As an approach to identify some of these inflammatory cytokines and the molecular mechanisms involved within different stages of hepatic IRI, we used an advanced antibody array assay to detect multiple proteins. With this technology, we observed specific differences in the content of inflammatory cytokines between ischemic and sham controls, as well as a function of the different reperfusion stages in a hepatic IRI mouse model. For example, while liver tissue content of IL-12p40/p70 was significantly increased in the ischemic stage, it was significantly decreased in the reperfusion stage as compared with that of the sham group. For other inflammatory cytokines, no changes were obtained between the ischemic and reperfusion stages with levels of IL-17, Eotaxin-2, Eotaxin, and sTNF RII all being consistently increased, while those of TIMP-1, TIMP-2, BLC, and MCSF consistently decreased as compared with that of the sham group at all reperfusion stages examined. Results from protein function annotation Gene Ontology and the KEGG pathway revealed that inflammatory cytokines are enriched in a network associated with activation of inflammatory response signaling pathways such as TLR, TNF, and IL-17 when comparing responses of the IR versus sham groups. The identification of cytokines along with their roles at specific stages of IRI may reveal important new biological markers for the diagnosis and prognosis of hepatic IRI.


hepatic ischemia-reperfusion injury inflammatory cytokines diagnosis and prognosis biomarker 



All the authors would like to thank the members of the Research Center of Medical College of Henan Polytechnic University, and Life Science Research Center of the First Affiliated Hospital of Xinxiang Medical University for their technical support.

Author Contributions

Li SP, Wang FF, and Zhang WK performed the majority of experiments and analyzed the data; Bian MZ and Zhang SY performed the molecular investigations; Li SP, Fang Y, Yan H, and Zhang WK participated equally in treatment of animals; Li SP and Zhang HM designed and coordinated the research; Li SP wrote the paper.

Funding Information

This work was supported by National Natural Science Foundation of China (No. 81700556).

Compliance with Ethical Standards

Conflict of Interest

The authors declare that they have no conflict of interest.


  1. 1.
    Li, J., R.J. Li, G.Y. Lv, and H.Q. Liu. 2015. The mechanisms and strategies to protect from hepatic ischemia-reperfusion injury. European Review for Medical and Pharmacological Sciences 19: 2036–2047.Google Scholar
  2. 2.
    Cannistra, M., M. Ruggiero, A. Zullo, G. Gallelli, S. Serafini, M. Maria, A. Naso, et al. 2016. Hepatic ischemia reperfusion injury: A systematic review of literature and the role of current drugs and biomarkers. International Journal of Surgery 33 (Suppl 1): S57–S70.CrossRefGoogle Scholar
  3. 3.
    Xi, J., M. Yan, S. Li, H. Song, L. Liu, Z. Shen, and J.Z. Cai. 2019. NOD1 activates autophagy to aggravate hepatic ischemia-reperfusion injury in mice. Journal of Cellular Biochemistry 120: 10605–10612.CrossRefGoogle Scholar
  4. 4.
    Sikalias, N., T. Karatzas, K. Alexiou, L. Mountzalia, M. Demonakou, I.D. Kostakis, A. Zacharioudaki, A. Papalois, and G. Kouraklis. 2018. Intermittent Ischemic Preconditioning Protects Against Hepatic Ischemia-Reperfusion Injury and Extensive Hepatectomy in Steatotic Rat Liver. Journal of Investigative Surgery 31: 366–377.CrossRefGoogle Scholar
  5. 5.
    Kan, C., L. Ungelenk, A. Lupp, O. Dirsch, and U. Dahmen. 2018. Ischemia-Reperfusion Injury in Aged Livers-The Energy Metabolism, Inflammatory Response, and Autophagy. Transplantation 102: 368–377.Google Scholar
  6. 6.
    Li, C.X., C.M. Lo, Q. Lian, K.T. Ng, X.B. Liu, Y.Y. Ma, X. Qi, et al. 2016. Repressor and activator protein accelerates hepatic ischemia reperfusion injury by promoting neutrophil inflammatory response. Oncotarget 7: 27711–27723.Google Scholar
  7. 7.
    Wu, H.H., C.C. Huang, C.P. Chang, M.T. Lin, K.C. Niu, and Y.F. Tian. 2018. Heat Shock Protein 70 (HSP70) Reduces Hepatic Inflammatory and Oxidative Damage in a Rat Model of Liver Ischemia/Reperfusion Injury with Hyperbaric Oxygen Preconditioning. Medical Science Monitor 24: 8096–8104.CrossRefGoogle Scholar
  8. 8.
    Ma, G., C. Chen, H. Jiang, Y. Qiu, Y. Li, X. Li, X. Zhang, J. Liu, and T. Zhu. 2017. Ribonuclease attenuates hepatic ischemia reperfusion induced cognitive impairment through the inhibition of inflammatory cytokines in aged mice. Biomedicine & Pharmacotherapy 90: 62–68.CrossRefGoogle Scholar
  9. 9.
    Matsumoto, T., K. O'Malley, P.A. Efron, C. Burger, P.F. McAuliffe, P.O. Scumpia, T. Uchida, et al. 2006. Interleukin-6 and STAT3 protect the liver from hepatic ischemia and reperfusion injury during ischemic preconditioning. Surgery 140: 793–802.CrossRefGoogle Scholar
  10. 10.
    Suyavaran, A., C. Ramamurthy, R. Mareeswaran, A. Subastri, R.P. Lokeswara, and C. Thirunavukkarasu. 2015. TNF-alpha suppression by glutathione preconditioning attenuates hepatic ischemia reperfusion injury in young and aged rats. Inflammation Research 64: 71–81.CrossRefGoogle Scholar
  11. 11.
    Kibat, J., T. Schirrmann, M.J. Knape, S. Helmsing, D. Meier, M. Hust, C. Schroder, et al. 2016. Utilisation of antibody microarrays for the selection of specific and informative antibodies from recombinant library binders of unknown quality. New Biotechnology 33: 574–581.CrossRefGoogle Scholar
  12. 12.
    Sun, X.L., Y.L. Zhang, S.M. Xi, L.J. Ma, and S.P. Li. 2019. MiR-330-3p suppresses phosphoglycerate mutase family member 5 -inducted mitophagy to alleviate hepatic ischemia-reperfusion injury. Journal of Cellular Biochemistry 120: 4255–4267.CrossRefGoogle Scholar
  13. 13.
    Qu, Y., H.L. Zhang, X.P. Zhang, and H.L. Jiang. 2018. Arachidonic acid attenuates brain damage in a rat model of ischemia/reperfusion by inhibiting inflammatory response and oxidative stress. Human & Experimental Toxicology 37: 135–141.CrossRefGoogle Scholar
  14. 14.
    Chen, J., D.M. Zhang, X. Feng, J. Wang, Y.Y. Qin, T. Zhang, Q. Huang, R. Sheng, Z. Chen, M. Li, and Z.H. Qin. 2018. TIGAR inhibits ischemia/reperfusion-induced inflammatory response of astrocytes. Neuropharmacology 131: 377–388.CrossRefGoogle Scholar
  15. 15.
    Zhao, D., S.C. Deng, Y. Ma, Y.H. Hao, and Z.H. Jia. 2018. miR-221 alleviates the inflammatory response and cell apoptosis of neuronal cell through targeting TNFAIP2 in spinal cord ischemia-reperfusion. Neuroreport 29: 655–660.Google Scholar
  16. 16.
    Ling, H., H. Chen, M. Wei, X. Meng, Y. Yu, and K. Xie. 2016. The Effect of Autophagy on Inflammation Cytokines in Renal Ischemia/Reperfusion Injury. Inflammation 39: 347–356.CrossRefGoogle Scholar
  17. 17.
    Gregova, K., S. Cikos, M. Bilecova-Rabajdova, P. Urban, J. Varga, S. Feterik, and J. Vesela. 2015. Intestinal ischemia-reperfusion injury mediates expression of inflammatory cytokines in rats. General Physiology and Biophysics 34: 95–99.CrossRefGoogle Scholar
  18. 18.
    Gu, L., Y. Tao, C. Chen, Y. Ye, X. Xiong, and Y. Sun. 2018. Initiation of the inflammatory response after renal ischemia/reperfusion injury during renal transplantation. International Urology and Nephrology 50: 2027–2035.CrossRefGoogle Scholar
  19. 19.
    Afkham, A., S. Eghbal-Fard, H. Heydarlou, R. Azizi, L. Aghebati-Maleki, and M. Yousefi. 2019. Toll-like receptors signaling network in pre-eclampsia: An updated review. Journal of Cellular Physiology 234: 2229–2240.CrossRefGoogle Scholar
  20. 20.
    Mohan, S., and D. Gupta. 2018. Crosstalk of toll-like receptors signaling and Nrf2 pathway for regulation of inflammation. Biomedicine & Pharmacotherapy 108: 1866–1878.CrossRefGoogle Scholar
  21. 21.
    Bahrami, A., N. Parsamanesh, S.L. Atkin, M. Banach, and A. Sahebkar. 2018. Effect of statins on toll-like receptors: a new insight to pleiotropic effects. Pharmacological Research 135: 230–238.CrossRefGoogle Scholar
  22. 22.
    Rao, J., S. Yue, Y. Fu, J. Zhu, X. Wang, R.W. Busuttil, J.W. Kupiec-Weglinski, L. Lu, and Y. Zhai. 2014. ATF6 mediates a pro-inflammatory synergy between ER stress and TLR activation in the pathogenesis of liver ischemia-reperfusion injury. American Journal of Transplantation 14: 1552–1561.CrossRefGoogle Scholar
  23. 23.
    Tsung, A., R.A. Hoffman, K. Izuishi, N.D. Critchlow, A. Nakao, M.H. Chan, M.T. Lotze, D.A. Geller, and T.R. Billiar. 2005. Hepatic ischemia/reperfusion injury involves functional TLR4 signaling in nonparenchymal cells. Journal of Immunology 175: 7661–7668.CrossRefGoogle Scholar
  24. 24.
    Chen, H., R.Q. Zhang, X.G. Wei, X.M. Ren, and X.Q. Gao. 2016. Mechanism of TLR-4/NF-kappaB pathway in myocardial ischemia reperfusion injury of mouse. Asian Pacific Journal of Tropical Medicine 9: 503–507.CrossRefGoogle Scholar
  25. 25.
    Perry, B.C., D. Soltys, A.H. Toledo, and L.H. Toledo-Pereyra. 2011. Tumor necrosis factor-alpha in liver ischemia/reperfusion injury. Journal of Investigative Surgery 24: 178–188.CrossRefGoogle Scholar
  26. 26.
    Jaeschke, H., and B.L. Woolbright. 2013. Role of heme oxygenase 1 in TNF/TNF receptor-mediated apoptosis after hepatic ischemia/reperfusion in rats. Shock 39: 380–388 Shock 2013;40:75–76.CrossRefGoogle Scholar
  27. 27.
    Spencer, N.Y., W. Zhou, Q. Li, Y. Zhang, M. Luo, Z. Yan, T.J. Lynch, D. Abbott, B. Banfi, and J.F. Engelhardt. 2013. Hepatocytes produce TNF-alpha following hypoxia-reoxygenation and liver ischemia-reperfusion in a NADPH oxidase- and c-Src-dependent manner. American Journal of Physiology. Gastrointestinal and Liver Physiology 305: G84–G94.CrossRefGoogle Scholar
  28. 28.
    Sharma, A.K., D.P. Mulloy, L.T. Le, and V.E. Laubach. 2014. NADPH oxidase mediates synergistic effects of IL-17 and TNF-alpha on CXCL1 expression by epithelial cells after lung ischemia-reperfusion. American Journal of Physiology. Lung Cellular and Molecular Physiology 306: L69–L79.CrossRefGoogle Scholar
  29. 29.
    Loi, P., Q. Yuan, D. Torres, S. Delbauve, M.A. Laute, M.C. Lalmand, M. Petein, et al. 2013. Interferon regulatory factor 3 deficiency leads to interleukin-17-mediated liver ischemia-reperfusion injury. Hepatology 57: 351–361.CrossRefGoogle Scholar
  30. 30.
    Wu, C., Y. Xia, P. Wang, L. Lu, and F. Zhang. 2011. Triptolide protects mice from ischemia/reperfusion injury by inhibition of IL-17 production. International Immunopharmacology 11: 1564–1572.CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Shi-peng Li
    • 1
    Email author
  • Fei-fei Wang
    • 2
  • Wen-kui Zhang
    • 3
  • Ming-ze Bian
    • 3
  • Shen-yan Zhang
    • 3
  • Han Yan
    • 3
  • Yuan Fang
    • 3
  • Hai-ming Zhang
    • 4
    Email author
  1. 1.Department of General Surgery, Jiaozuo People’s HospitalXinxiang Medical UniversityJiaozuoPeople’s Republic of China
  2. 2.Department of Geriatric Medicine, Jiaozuo People’s HospitalXinxiang Medical UniversityJiaozuoPeople’s Republic of China
  3. 3.Department of PharmacyMedical School of Henan Polytechnic UniversityJiaozuoPeople’s Republic of China
  4. 4.Liver Transplantation Center, Beijing Friendship HospitalCapital Medical UniversityBeijingPeople’s Republic of China

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