Abstract:
Background:Ischemia-reperfusion (IR) is a major contributor to graft rejection after liver transplantation. During IR injury, an intense inflammatory process occurs in the liver. Neutrophils are considered central players in the events that lead to liver injury. CXC chemokines mediate hepatic inflammation following reperfusion. However, few studies have demonstrated in real-time the behavior of recruited neutrophils. We used confocal intravital microscopy (IVM) to image neutrophil migration in the liver and to analyze in real-time parameters of neutrophil recruitment in the inflamed tissue in animals treated or not with reparixin, an allosteric antagonist of CXCR1/2 receptors. Materials and methods:WT and LysM-eGFP mice treated with reparixin or saline were subjected to 60 min of ischemia followed by different times of reperfusion. Mice received Sytox orange intravenously to show necrotic DNA in IVM. The effect of reparixin on parameters of local and systemic reperfusion-induced injury was also investigated. Results:IR induced liver injury and inflammation, as evidenced by high levels of alanine aminotransferase and myeloperoxidase activity, chemokine and cytokine production, and histological outcome. Treatment with reparixin significantly decreased neutrophil influx. Moreover, reparixin effectively suppressed the increase in serum concentrations of TNF-α, IL-6, and CCL3, and the reperfusion-associated tissue damage. The number of neutrophils in the liver increased between 6 and 24 h of reperfusion, whereas the distance traveled, velocity, neutrophil size and shape, and cluster formation reached a maximum 6 h after reperfusion and then decreased gradually. In vivo imaging revealed that reparixin significantly decreased neutrophil infiltration and movement and displacement of recruited cells. Moreover, neutrophils had a smaller size and less elongated shape in treated mice. Conclusion:Imaging of the liver by confocal IVM was successfully implemented to describe neutrophil behavior in vivo during liver injury by IR. Treatment with reparixin decreased not only the recruitment of neutrophils in tissues but also their activation state and capacity to migrate within the liver. CXCR1/2 antagonists may be a promising therapy for patients undergoing liver transplantation.
journal_name
Front Immunoljournal_title
Frontiers in immunologyauthors
de Oliveira THC,Marques PE,Poosti F,Ruytinx P,Amaral FA,Brandolini L,Allegretti M,Proost P,Teixeira MMdoi
10.3389/fimmu.2017.01917subject
Has Abstractpub_date
2018-01-05 00:00:00pages
1917issn
1664-3224journal_volume
8pub_type
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pub_type: 杂志文章,已发布勘误
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journal_title:Frontiers in immunology
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pub_type: 杂志文章
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pub_type: 临床试验,杂志文章
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journal_title:Frontiers in immunology
pub_type: 杂志文章
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更新日期:2018-04-30 00:00:00
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journal_title:Frontiers in immunology
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journal_title:Frontiers in immunology
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pub_type: 杂志文章,评审
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