Expression of pro-inflammatory genes in lesions, spleens and blood neutrophils after burn injuries in mice treated with silver sulfodiazine

Document Type : Original Article


1 Department of Pathobiology, Faculty of Veterinary Science, Bu-Ali Sina University, Hamedan, Iran

2 Department of Clinical Sciences, Faculty of Veterinary Science, Hamedan, Iran


Objective(s): It is now supposed that cytokines released during the burn injuries have a great impact on the immunological and pathological responses after the burn. The main objective of this study was to investigate the expression of some pro-inflammatory genes in the wound, spleen and blood neutrophils during the healing process of burn wounds in a murine model.
Materials and Methods: The expression of ten pro-inflammatory genes were examined in wounds, spleens and blood neutrophils of mice with burn injuries treated with either silver sulfodiazine or phosphate-buffered saline (PBS) using RT-PCR at the end of the first and second weeks after injuries.
Results: None of the pro-inflammatory genes were expressed in the skin, spleen and blood neutrophils of healthy mice. In the group control, IL-12P35, IL-12P40, CCR5, IL-1β and IFN- γ were expressed in the spleen and blood neutrophils in the first week. Instead, CCL5, CCR5, IL-1β and IFN- γ were expressed in the wound, but in the second week, the expression of the genes became similar. In the test group, in the first week, TNF-α, IL-12P35, IL-12P40 and IL-1β were expressed in the lesions, CCL4, IL-1α, IL-12P35, IL-12P40, CCR5 and IFN- γ were expressed in the spleen and no pro-inflammatory gene expression was detected in blood neutrophils. 
Conclusion: IL-1β and IFN- γ are expressed in wound, spleen and neutrophils of untreated mice, but not in silver sulfodiazine treated mice. Hence, treatment with silver sulfodiazine suppressed the expression of pro-inflammatory genes in some stages of healing.


1. Feezor RJ, Paddock HN, Baker HV, Varela JC, Barreda J, Moldawer LL, et al. Temporal patterns of gene expression in murine cutaneous burn wound healing. Physiol Genom 2004; 16:341-348.
2. Shakespeare P. Burn wound healing and skin substitutes. Burns 2001; 27:517-522.
3. Linares H. From wound to scar. Burns 1996; 22:339-352.
4. Gamelli RL, He L-k, Liu H. Marrow granulocyte-macrophage progenitor cell response to burn injury as modified by endotoxin and indomethacin. J Trauma  Acute Care Surg 1994; 37:339-346.
5. Hansbrough JF, Zapata-Sirvent R, Peterson V, Wang X, Bender E, Claman H, et al. Characterization of the immunosuppressive effect of burned tissue in an animal model. J Surg Res 1984; 37:383-393.
6. Miller CL, Baker CC. Changes in lymphocyte activity after thermal injury: The role of suppressor cells. J Clin Invest 1979; 63:202.
7. Herndon DN, Wilmore DW, Mason AD. Development and analysis of a small animal model simulating the human postburn hypermetabolic response. J Surg Res 1978; 25:394-403.
8. Kataranovski M, Magic Z, Pejnovic N. Early inflammatory cytokine and acute phase protein response under the stress of thermal injury in rats. Physiol Res 1999; 48:473-482.
9. Zimmermann M. Ethical guidelines for investigations of experimental pain in conscious animals. Pain 1983; 16:109-110.
10. Cribbs RK, Luquette MH, Besner GE. A standardized model of partial thickness scald burns in mice. J Surg Res 1998; 80:69-74.
11. Rezapour A. The neutrophiwere isolated according to the method explained by Rezapour with modifications. J Anim Vet Adv 2009; 8:11-15.
12. Rezvan H. Molecular cloning of leishmania major gp63 Gene in BALB/c Mouse CT26 Cell Line. Zahedan J Res Med Sci 2014; 17:5-9.
13. Sacks D, Noben-Trauth N. The immunology of susceptibility and resistance to Leishmania major in mice. Nat Rev Immunol 2002; 2:845-858.
14. Inatsu A, Kogiso M, Jeschke MG, Asai A, Kobayashi M, Herndon DN, et al. Lack of Th17 cell generation in patients with severe burn injuries. J Immunol 2011; 187:2155-2161.
15. Gragnani A, Cezillo MV, da Silva ID, de Noronha SM, Correa-Noronha SA, Ferreira LM. Gene expression profile of cytokines and receptors of inflammation from cultured keratinocytes of burned patients. Burns 2014; 40:947-956.
16. Hultman CS, Napolitano LM, Cairns BA, Brady LA, Campbell C, deSerres S, et al. The relationship between interferon-gamma and keratinocyte alloantigen expression after burn injury. Ann Surg 1995; 222:384-389; discussion 392-383.
17. Huber NL, Bailey SR, Schuster R, Ogle CK, Lentsch AB, Pritts TA. Prior thermal injury accelerates endotoxin-induced inflammatory cytokine production and intestinal nuclear factor-κB activation in mice. J Burn Care Res 2012; 33:279-285.
18. Gauglitz GG, Zedler S, von Spiegel F, Fuhr J, von Donnersmarck GH, Faist E. Functional characterization of cultured keratinocytes after acute cutaneous burn injury. PLoS One 2012; 7:e29942.
19. Cannon JG, Friedberg JS, Gelfand JA, Tompkins RG, Burke JF, Dinarello CA. Circulating interleukin-1 beta and tumor necrosis factor-alpha concentrations after burn injury in humans. Crit Care Med 1992; 20:1414-1419.
20. Gragnani A, Muller BR, Silva ID, Noronha SM, Ferreira LM. Keratinocyte growth factor, tumor necrosis factor-alpha and interleukin-1 beta gene expression in cultured fibroblasts and keratinocytes from burned patients. Acta Cir Bras 2013; 28:551-558.
21. Mandrup-Poulsen T, Wogensen LD, Jensen M, Svensson P, Nilsson P, Emdal T, et al. Circulating interleukin-1 receptor antagonist concentrations are increased in adult patients with thermal injury. Crit Care Med 1995; 23:26-33.
22. Tang A, Gilchrest BA. Regulation of keratinocyte growth factor gene expression in human skin fibroblasts. J Dermatol Sci 1996; 11:41-50.
23. Spees AM, Kingsbury DD, Wangdi T, Xavier MN, Tsolis RM, Bäumler AJ. Neutrophils are a source of gamma interferon during acute Salmonella enterica serovar Typhimurium colitis. Infect Immun 2014; 82:1692-1697.
24. Sturge CR, Benson A, Raetz M, Wilhelm CL, Mirpuri J, Vitetta ES, et al. TLR-independent neutrophil-derived IFN-γ is important for host resistance to intracellular pathogens. Proc Natl Acad Sci 2013; 110:10711-10716.
25. Rodrigues DR, Fernandes RK, de Almeida Balderramas H, Penitenti M, Bachiega TF, Calvi SA, et al. Interferon-gamma production by human neutrophils upon stimulation by IL-12, IL-15 and IL-18 and challenge with Paracoccidioides brasiliensis. Cytokine 2014; 69:102-109.
26. Lund JM, Alexopoulou L, Sato A, Karow M, Adams NC, Gale NW, et al. Recognition of single-stranded RNA viruses by Toll-like receptor 7. Proc Natl Acad Sci U S A 2004; 101:5598-5603.
27. Cai S, Batra S, Langohr I, Iwakura Y, Jeyaseelan S. IFN-γ induction by neutrophil-derived IL-17A homodimer augments pulmonary antibacterial defense. Mucosal Immunol 2016; 9:718-292015.
28. Haverkamp MH, van de Vosse E, van Dissel JT. Nontuberculous mycobacterial infections in children with inborn errors of the immune system. J Infect 2014; 68:S134-S150.
29. Balderramas HA, Penitenti M, Rodrigues DR, Bachiega TF, Fernandes RK, Ikoma MRV, et al. Human neutrophils produce IL-12, IL-10, PGE2 and LTB4 in response to Paracoccidioides brasiliensis. involvement of TLR2, mannose receptor and dectin-1. Cytokine 2014; 67:36-43.