pp 1–14 | Cite as

Visfatin Exerts Immunotherapeutic Effects in Lipopolysaccharide-Induced Acute Lung Injury in Murine Model

  • You Luo
  • Xin-Xin Pang
  • Abdur Rahman Ansari
  • Xin-Tong Wu
  • Hui-Zhen Li
  • Zhe-Wei Zhang
  • Hui SongEmail author
Original Article


Visfatin acts as a significant regulator of inflammatory cytokines. However, the immunological response and therapeutic effects of visfatin under bacterial stress in murine lung tissue are still not clear. To investigate the role of visfatin on lipopolysaccharide (LPS)-induced acute lung injury (ALI), thirty Kunming mice were divided into Saline, LPS, and LPS + visfatin groups. After routine blood examination, the effects of visfatin on inflammatory cytokines, lung tissue structure, and expression of inflammatory mediators were explored through hematoxylin-eosin (H&E), Masson and immunohistochemical staining, quantitative polymerase chain reaction (Q-PCR), and Western blotting. Compared with the Saline group, neutrophil percentage, peripheral blood neutrophil count, and the ratio of lymphocyte count (NLR) were upregulated in LPS group. Moreover, Masson staining showed alterations in lung tissue structure; the mRNA level of different cytokines (IL-6, IL-1β, TNF-α, IL-10, TLR4, IFN-γ) was upregulated; and the protein expression of interleukin (IL)-6, myeloperoxidase (MPO), and transforming growth factor-β1 (TGF-β) was significantly (p < 0.05) different in LPS group. Compared with LPS group, neutrophil percentage significantly decreased (p < 0.01), the numbers of lymphocytes significantly (p < 0.05) increased, NLR decreased, Masson staining of the lung was extremely different (p < 0.01), the structure of the lung was slightly damaged, and the myeloperoxidase values of lung showed no differences in LPS + visfatin. Hence, visfatin inhibits the lung inflammation induced by ALI. During the ALI, visfatin acts by decreasing NLR, downregulated the expression of MPO, enhanced antioxidant capacity, and regulated the inflammatory factors IL-1β, IL-6, IL-10, and TNF-α to reduce the lung injury.


acute lung injury inflammatory cytokines lipopolysaccharide visfatin 


Funding Information

This study was supported by National Natural Science Fund Project of China (No. 31772687), Fundamental Research Funds for the Central Universities (No. 2662015PY063), and National Natural Science Fund Project of China (No. 31101776).

Compliance with Ethical Standards

The current study was performed in accordance with the guidelines of the Animal Care and Use Committee of Huazhong Agricultural University (HZAU), Wuhan, China.

Conflict of Interest

The authors declare that they have no conflict of interest.


  1. 1.
    Apostolopoulos, V., M.P. de Courten, L. Stojanovska, G.L. Blatch, K. Tangalakis, and B. de Courten. 2016. The complex immunological and inflammatory network of adipose tissue in obesity. Molecular Nutrition & Food Research 60 (1): 43–57.CrossRefGoogle Scholar
  2. 2.
    Bartram, Ulrike, and Christian P. Speer. 2004. The role of transforming growth factor β in lung development and disease. Chest 125 (2): 754–765. Scholar
  3. 3.
    Buyukkaya, Eyup, Mehmet Fatih Karakaş, Esra Karakaş, Adnan Burak Akçay, Ibrahim Halil Tanboga, Mustafa Kurt, and Nihat Sen. 2014. Correlation of neutrophil to lymphocyte ratio with the presence and severity of metabolic syndrome. Clinical and Applied Thrombosis/Hemostasis 20 (2): 159–163. Scholar
  4. 4.
    Cen, Cindy, Monowar Aziz, and Ping Wang. 2017. Novel inflammatory and immunomodulatory mediators in sepsis. In Advanced trauma and surgery, ed. Fu Xiaobing and Liangming Liu, 211–234. Singapore: Springer Singapore.CrossRefGoogle Scholar
  5. 5.
    Chen, Tong, Qianqian Guo, Huimin Wang, Huixin Zhang, Ciman Wang, Ping Zhang, Shanshan Meng, Yunman Li, Hui Ji, and Tianhua Yan. 2015. Effects of esculetin on lipopolysaccharide (LPS)-induced acute lung injury via regulation of RhoA/Rho kinase/NF-кB pathways in vivo and in vitro. Free Radical Research 49 (12): 1459–1468.CrossRefGoogle Scholar
  6. 6.
    Chen, Tong, Yi Mou, Jiani Tan, Linlin Wei, Yixue Qiao, Tingting Wei, Pengjun Xiang, Sixun Peng, Yihua Zhang, and Zhangjian Huang. 2015. The protective effect of CDDO-Me on lipopolysaccharide-induced acute lung injury in mice. International Immunopharmacology 25 (1): 55–64.CrossRefGoogle Scholar
  7. 7.
    Chesnutt, Asha N., Michael A. Matthay, Fred A. Tibayan, and Joan G. Clark. 1997. Early detection of type III procollagen peptide in acute lung injury: pathogenetic and prognostic significance. American Journal of Respiratory and Critical Care Medicine 156 (3): 840–845.CrossRefGoogle Scholar
  8. 8.
    Crapo, James D. 1986. Morphologic changes in pulmonary oxygen toxicity. Annual Review of Physiology 48 (1): 721–731.CrossRefGoogle Scholar
  9. 9.
    Davies, Michael J. 2011. Myeloperoxidase-derived oxidation: mechanisms of biological damage and its prevention. Journal of Clinical Biochemistry and Nutrition 48 (1): 8–19. Scholar
  10. 10.
    Díaz, Fernando Erra, Ezequiel Dantas, Maia Cabrera, Constanza A. Benítez, María V. Delpino, Gabriel Duette, Julia Rubione, Norberto Sanjuan, Analía S. Trevani, and Jorge Geffner. 2016. Fever-range hyperthermia improves the anti-apoptotic effect induced by low pH on human neutrophils promoting a proangiogenic profile. Cell Death &Amp; Disease 7: e2437.
  11. 11.
    dos Santos, Meyer, Karina Kuczka Sascha, Bettina Picard-Willems, Karen Nelson, Ute Klinkhardt, and Sebastian Harder. 2015. The integrin antagonist, cilengitide, is a weak inhibitor of αIIbβ3 mediated platelet activation and inhibits platelet adhesion under flow. Platelets 26 (1): 59–66.CrossRefGoogle Scholar
  12. 12.
    Farivar, Alexander S., Baiya Krishnadasan, Babu V. Naidu, Steven M. Woolley, Edward D. Verrier, and Michael S. Mulligan. 2003. Endogenous interleukin-4 and interleukin-10 regulate experimental lung ischemia reperfusion injury. The Annals of Thoracic Surgery 76 (1): 253–259.CrossRefGoogle Scholar
  13. 13.
    Feist, Peter E., Elizabeth A. Loughran, M. Sharon Stack, and Amanda B. Hummon. 2018. Quantitative proteomic analysis of murine white adipose tissue for peritoneal cancer metastasis. Analytical and Bioanalytical Chemistry 410 (5): 1583–1594. Scholar
  14. 14.
    Feng, Xiaosheng, and Aiqing Jia. 2014. Protective effect of carvacrol on acute lung injury induced by lipopolysaccharide in mice. Inflammation 37 (4): 1091–1101.CrossRefGoogle Scholar
  15. 15.
    Fontana, Luigi, J. Christopher Eagon, Maria E. Trujillo, Philipp E. Scherer, and Samuel Klein. 2007. Visceral fat adipokine secretion is associated with systemic inflammation in obese humans. Diabetes 56 (4): 1010–1013. Scholar
  16. 16.
    Frank, James A., Polly E. Parsons, and Michael A. Matthay. 2006. Pathogenetic significance of biological markers of ventilator-associated lung injury in experimental and clinical studies. Chest Journal 130 (6): 1906–1914.CrossRefGoogle Scholar
  17. 17.
    Griet, Milagros, Hortensia Zelaya, Melina Valeria Mateos, Susana Salva, Guillermo Esteban Juarez, Graciela Font de Valdez, Julio Villena, Gabriela Alejandra Salvador, and Ana Virginia Rodriguez. 2014. Soluble factors from Lactobacillus reuteri CRL1098 have anti-inflammatory effects in acute lung injury induced by lipopolysaccharide in mice. PLoS One 9 (10): e110027.CrossRefGoogle Scholar
  18. 18.
    Hart, Kevin M., Thomas Fabre, Joshua C. Sciurba, Richard L. Gieseck, Lee A. Borthwick, Kevin M. Vannella, Thomas H. Acciani, Rafael de Queiroz Prado, Robert W. Thompson, and Sandra White. 2017. Type 2 immunity is protective in metabolic disease but exacerbates NAFLD collaboratively with TGF-β. Science Translational Medicine 9 (396): eaal3694.CrossRefGoogle Scholar
  19. 19.
    Havixbeck, Jeffrey J., Aja M. Rieger, Michael E. Wong, Jordan W. Hodgkinson, and Daniel R. Barreda. 2016. Neutrophil contributions to the induction and regulation of the acute inflammatory response in teleost fish. Journal of Leukocyte Biology 99 (2): 241–252. Scholar
  20. 20.
    Hinz, Boris. 2015. The extracellular matrix and transforming growth factor-β1: tale of a strained relationship. Matrix Biology 47: 54–65. Scholar
  21. 21.
    Hong, Kyunghee, Guanlan Xu, Truman B Grayson, and Anath Shalev. 2016. Cytokines regulate β-cell TXNIP via distinct mechanisms and pathways. Journal of Biological Chemistry:jbc. M115. 698365.Google Scholar
  22. 22.
    Kalaiyarasu, Semmannan, Sandeep Bhatia, Niranjan Mishra, Richa Sood, Kumar Manoj, D. SenthilKumar, Sushant Bhat, and M. Dass Prakash. 2016. Elevated level of pro inflammatory cytokine and chemokine expression in chicken bone marrow and monocyte derived dendritic cells following LPS induced maturation. Cytokine 85: 140–147.CrossRefGoogle Scholar
  23. 23.
    Khan, Amjad, Mohammed Alsahli, and Arshad Rahmani. 2018. Myeloperoxidase as an active disease biomarker: recent biochemical and pathological perspectives. Medical Science 6 (2): 33.Google Scholar
  24. 24.
    Koch, Alexander, Ralf Weiskirchen, Alexander Krusch, Jan Bruensing, Lukas Buendgens, Ulf Herbers, Eray Yagmur, Ger H. Koek, Christian Trautwein, and Frank Tacke. 2018. Visfatin serum levels predict mortality in critically ill patients. Disease Markers 2018: 8. Scholar
  25. 25.
    Kolaczkowska, Elzbieta, and Paul Kubes. 2013. Neutrophil recruitment and function in health and inflammation. Nature Reviews Immunology 13: 159. Scholar
  26. 26.
    Liu, Fang, Guo-quan Sun, Hua-yi Gao, Rui-sheng Li, Lanan-Wassy Soromou, Na Chen, Yan-Hong Deng, and Hai-hua Feng. 2013. Angelicin regulates LPS-induced inflammation via inhibiting MAPK/NF-κB pathways. Journal of Surgical Research 185 (1): 300–309.CrossRefGoogle Scholar
  27. 27.
    Livak, Kenneth J., and Thomas D. Schmittgen. 2001. Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCT method. Methods 25 (4): 402–408.CrossRefGoogle Scholar
  28. 28.
    Luk, Tracy, Zeenat Malam, and John C. Marshall. 2008. Pre-B cell colony-enhancing factor (PBEF)/visfatin: a novel mediator of innate immunity. Journal of Leukocyte Biology 83 (4): 804–816.CrossRefGoogle Scholar
  29. 29.
    MacKinnon, Alison C., Michael A. Gibbons, Sarah L. Farnworth, Hakon Leffler, Ulf J. Nilsson, Delaine Tamara, A. John Simpson, et al. 2012. Regulation of transforming growth factor-β1–driven lung fibrosis by galectin-3. American Journal of Respiratory and Critical Care Medicine 185 (5): 537–546. Scholar
  30. 30.
    Miller, Hugh R.P., and Alan D. Pemberton. 2002. Tissue-specific expression of mast cell granule serine proteinases and their role in inflammation in the lung and gut. Immunology 105 (4): 375–390.CrossRefGoogle Scholar
  31. 31.
    Moloney, E.D., and M.J.D. Griffiths. 2004. Protective ventilation of patients with acute respiratory distress syndrome. British Journal of Anaesthesia 92 (2): 261–270.CrossRefGoogle Scholar
  32. 32.
    Okuda, Suguru, David J. Sherman, Thomas J. Silhavy, Natividad Ruiz, and Daniel Kahne. 2016. Lipopolysaccharide transport and assembly at the outer membrane: the PEZ model. Nature Reviews Microbiology 14: 337. Scholar
  33. 33.
    Ozcelik, Fatih, Celalettin Yuksel, Erol Arslan, Sema Genc, Beyhan Omer, and Muhittin A. Serdar. 2013. Relationship between visceral adipose tissue and adiponectin, inflammatory markers and thyroid hormones in obese males with hepatosteatosis and insulin resistance. Archives of Medical Research 44 (4): 273–280.CrossRefGoogle Scholar
  34. 34.
    Pardo, Annie, Sandra Cabrera, Mariel Maldonado, and Moisés Selman. 2016. Role of matrix metalloproteinases in the pathogenesis of idiopathic pulmonary fibrosis. Respiratory Research 17 (1): 23.CrossRefGoogle Scholar
  35. 35.
    Parekh, Dhruv, Rachel C. Dancer, and David R. Thickett. 2011. Acute lung injury. Clinical Medicine 11 (6): 615–618. Scholar
  36. 36.
    Ray, Amitabha. 2012. Adipokine leptin in obesity-related pathology of breast cancer. Journal of Biosciences 37 (2): 289–294.CrossRefGoogle Scholar
  37. 37.
    Reddy, Narsa M., Vegiraju Suryanaraya, Melinda S. Yates, Steven R. Kleeberger, Paul M. Hassoun, Masayuki Yamamoto, Karen T. Liby, Michael B. Sporn, Thomas W. Kensler, and Sekhar P. Reddy. 2009. The triterpenoid CDDO-imidazolide confers potent protection against hyperoxic acute lung injury in mice. American Journal of Respiratory and Critical Care Medicine 180 (9): 867–874.CrossRefGoogle Scholar
  38. 38.
    Schwechheimer, Carmen, and Meta J. Kuehn. 2015. Outer-membrane vesicles from Gram-negative bacteria: biogenesis and functions. Nature Reviews Microbiology 13: 605. Scholar
  39. 39.
    Shiny, Abhijit, Yesodha S. Bibin, Coimbatore Subramanian Shanthirani, Bhaskaran S. Regin, Ranjit Mohan Anjana, Muthuswamy Balasubramanyam, Saravanan Jebarani, and Viswanathan Mohan. 2014. Association of neutrophil-lymphocyte ratio with glucose intolerance: an indicator of systemic inflammation in patients with type 2 diabetes. Diabetes Technology & Therapeutics 16 (8): 524–530.CrossRefGoogle Scholar
  40. 40.
    Singh, Manindra, and Fabian Benencia. 2017. Inflammatory signals from fat: visfatin promotes pro-inflammatory activation and leukocyte interaction in endothelial cells. American Association of Immunology.Google Scholar
  41. 41.
    Tellides, George, and Jordan S. Pober. 2015. Inflammatory and immune responses in the arterial media. Circulation Research 116 (2): 312–322.CrossRefGoogle Scholar
  42. 42.
    Uhl, Bernd, Yannick Vadlau, Gabriele Zuchtriegel, Katharina Nekolla, Kariem Sharaf, Florian Gaertner, Steffen Massberg, Fritz Krombach, and Christoph A. Reichel. 2016. Aged neutrophils contribute to the first line of defense in the acute inflammatory response. Blood. Scholar
  43. 43.
    van der Poll, Tom, Frank L. van de Veerdonk, Brendon P. Scicluna, and Mihai G. Netea. 2017. The immunopathology of sepsis and potential therapeutic targets. Nature Reviews Immunology 17 (7): 407.CrossRefGoogle Scholar
  44. 44.
    Wei, Ying, Thomas J. Kim, David H. Peng, Dana Duan, Don L. Gibbons, Mitsuo Yamauchi, Julia R. Jackson, Claude J. Le Saux, Cheresa Calhoun, and Jay Peters. 2017. Fibroblast-specific inhibition of TGF-β1 signaling attenuates lung and tumor fibrosis. The Journal of Clinical Investigation 127 (10): 3675–3688.CrossRefGoogle Scholar
  45. 45.
    Wu, Xin-Tong, Zhi Yang, Abdur Rahman Ansari, Ke Xiao, Xin-Xin Pang, You Luo, and Hui Song. 2018. Visfatin regulates the production of lipopolysaccharide-induced inflammatory cytokines through p38 signaling in murine macrophages. Microbial Pathogenesis 117: 55–59.CrossRefGoogle Scholar
  46. 46.
    Wyllie, D.H., I.C.J.W. Bowler, and T.E.A. Peto. 2004. Relation between lymphopenia and bacteraemia in UK adults with medical emergencies. Journal of Clinical Pathology 57 (9): 950–955.CrossRefGoogle Scholar
  47. 47.
    Xiao, Ke, Wei-Hua Zou, Zhi Yang, Zia ur Rehman, Abdur Rahman Ansari, Huai-Rui Yuan, Ying Zhou, Lu Cui, Ke-Mei Peng, and Hui Song. 2015. The role of visfatin on the regulation of inflammation and apoptosis in the spleen of LPS-treated rats. Cell and Tissue Research 359 (2): 605–618.CrossRefGoogle Scholar
  48. 48.
    Xu, Haiyan, Glenn T. Barnes, Qing Yang, Tan Guo, Daseng Yang, Chieh J. Chou, Jason Sole, et al. 2003. Chronic inflammation in fat plays a crucial role in the development of obesity-related insulin resistance. Journal of Clinical Investigation 112 (12): 1821–1830. Scholar
  49. 49.
    Yang, Zhi, Ke Xiao, Wei Wang, Juan Tang, Peng-Peng Sun, Ke-Mei Peng, Z.Y.A.N.G. Hui Song, K. Xiao, and W. Wang. 2015. The effect of visfatin on inflammatory reaction in uterus of LPS-induced rats. International Journal of Morphology 33 (1): 194–203.CrossRefGoogle Scholar
  50. 50.
    Yen, Yu-Ting, Hong-Ren Yang, Hung-Chieh Lo, Ya-Ching Hsieh, Shih-Chang Tsai, Chia-Wen Hong, and Chi-Hsun Hsieh. Enhancing autophagy with activated protein C and rapamycin protects against sepsis-induced acute lung injury. Surgery 153 (5): 689–698. Scholar
  51. 51.
    Yu, Shouli, Min Shi, Changting Liu, Qinghui Liu, Jun Guo, Senyang Yu, and Tingshu Jiang. 2015. Time course changes of oxidative stress and inflammation in hyperoxia-induced acute lung injury in rats. Iranian Journal of Basic Medical Sciences 18 (1): 98–103.PubMedPubMedCentralGoogle Scholar
  52. 52.
    Yun, Mi Ran, Jeong Mi Seo, and Hyun Young Park. 2014. Visfatin contributes to the differentiation of monocytes into macrophages through the differential regulation of inflammatory cytokines in THP-1 cells. Cellular Signalling 26 (4): 705–715.CrossRefGoogle Scholar
  53. 53.
    Zhou, Ying, Cui Lu, Huairui Yuan, Ke Xiao, Faheem Ahmed Khan, Guo Liang, Zhi Yang, and Hui Song. 2015. Effect of visfatin on the structure and immune levels in the small intestine of LPS-induced rats. International Journal of Morphology 33 (4).CrossRefGoogle Scholar

Copyright information

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

Authors and Affiliations

  1. 1.College of Animal Science and Veterinary MedicineHuazhong Agricultural UniversityWuhanChina
  2. 2.Section of Anatomy and Histology, Department of Basic SciencesCollege of Veterinary and Animal Sciences (CVAS), JhangJhangPakistan
  3. 3.University of Veterinary and Animal Sciences (UVAS)LahorePakistan

Personalised recommendations