Abstract:Objective To investigate the role and significance of cardiac mast cells and Toll-like receptor 4 (TLR4) in the development and progression of viral myocarditis (VMC). Methods Forty-eight Balb/c mice were randomly divided into a control group (n=24) and a model group (n=24). Coxsackievirus B3 was intraperitoneally injected into the model group mice to establish a VMC model. In each group, cardiac tissues were collected from 8 mice at 7, 14 and 28 days after the model was established. The cardiac tissues were stained with hematoxylin and eosin as well as Masson trichrome to observe pathological changes in cardiac tissues. The number and degranulation of cardiac mast cells at each time point were measured and evaluated by toluidine blue staining and transmission electron microscopy. The mRNA and protein expression of TLR4 in cardiac tissues was measured by RT-PCR and immunohistochemistry. In the model group, the correlation between number of cardiac mast cells and mRNA expression of TLR4 at all time points was analyzed. Results The model group had significantly higher pathological scores of cardiac tissues than the control group at all time points (P<0.05). The myocardial collagen volume fraction in the model group at 28 days was significantly higher than in the control group at all time points and higher than in the model group at 7 and 14 days (P<0.05). At each time point, the model group had a significantly increased number of mast cells (P<0.05), and significantly increased mRNA and protein expression of TLR4 (P<0.05) compared with the control group. In the model group, the number of cardiac mast cells was positively correlated with the mRNA expression of TLR4 at all time points (R2=0.877, P<0.05). Conclusions Mice with VMC have significantly increased numbers of cardiac mast cells and expression of TLR4 compared with control mice at all time points, suggesting that mast cells and TLR4 may play important roles in the inflammatory response and fibrosis of VMC.
LI Hui,HUANG Lin-Feng,WEN Chun et al. Roles of cardiac mast cells and Toll-like receptor 4 in viral myocarditis among mice[J]. CJCP, 2013, 15(10): 896-902.
Fairweather D, Rose NR. Coxsackievirusinduced myocarditis in mice a model of autoimmune disease for studying immunotoxicity[J]. Methods, 2007, 41(1): 118-122.
[4]
Haeberle HA, Takizawa R, Casola A, Brasier AR, Dieterich HJ, Van Rooijen N, et al. Respiratory syncytial virus-induced activation of nuclear factor-kappa B in the lung involves alveolar macrophages and toll-like receptor 4-dependent pathways[J]. Infect Dis, 2002, 186(9): 1199-1206.
Goren N, Cuenca J, Martin-Sanz P, Bosca L. Attenuation of NF-kappa B signalling in rat cardiomyocytes at birth restricts the induction of inflammatory genes[J]. Cardiovasc Res, 2004, 64(2): 289-297.
[7]
Fairweather D, Yusung S, Frisancho S, Barrett M, Gatewood S, Steele R, et al. IL-12 receptor β1 and Toll-like receptor 4 increase IL-1β- and IL-18-associated myocarditis and coxsac-kievirus replication[J]. J Immunol, 2003, 170(9): 4731-4737.
[8]
Hirokazu H, Tadashi M, Makoto K, Tasuku Y, Koji U, Akira M. Roles of mast cells in the pathogenesis of viral myocarditis; reconstitution of mast cells to their deficient mice[J]. J Card Fail, 2004, 10(5): 162.
[9]
Ohtsuka T. Different interaction of mast cells with human endothelial cells and fibroblasts[J]. Eur J Dermatol, 2000, 10(2): 115-121.
[10]
Kitaura-Inenaga K, Hara M, Higuchi K, Yamamoto K, Yamaki A, Ono K, et al. Gene expression of cardiac mast cell chymase and tryptase in a murine model of heart failure caused by viral myocarditis[J]. Circ J, 2003, 67(10): 881-884.
[11]
Levick SP, Gardner JD, Holland M, Hauer-Jensen M, Janicki JS, Brower GL. Protection from adverse myocardial remodeling secondary to chronic volume overload in mast cell deficient rats[J]. J Mol Cell Cardiol, 2008, 45 (1): 56-61.
Fukui A, Ohta K, Nishi H, Shigeishi H, Tobiume K, Takechi M, et al. Interleukin-8 and CXCL10 expressions in oral keratinocytes and fibroblasts via Toll-like receptors[J]. Microbiol Immunol, 2013, 57(3): 198-206.
Dietrich N, Rohde M, Geffers R, Kroger A, Hauser H, Weiss S, et al. Mast cells elicit proinflammatory but not type I interferon responses upon activation of TLRs by bacteria[J]. Pro Natl Acad Sci USA, 2010, 107(19): 8748-8753.