Role of the thrombin/protease-activated receptor 1 pathway in intestinal ischemia-reperfusion injury in rats

Abstract

CXC chemokines, including human interleukin-8 and rat cytokine-induced neutrophil chemoattractant-1, play a crucial role in the pathogenesis of intestinal inflammation induced by ischemia-reperfusion (I-R). Thrombin and its specific receptor, protease-activated receptor 1 (PAR1), act as important players in inflammation. However, the association between thrombin activation and chemokine production during I-R has not been well studied. We investigated whether thrombin and PAR1 might be involved in the pathophysiology of intestinal I-R, using an in vivo model. Intestinal damage was induced by clamping the superior mesenteric artery for 30 min followed by reperfusion in male Wistar rats. Thrombin-antithrombin complex was measured as an indicator of thrombin activation. PAR1 expression in the intestine was evaluated by real-time PCR. The severity of the intestinal mucosal injury was evaluated on the distal segment of the ileum by several biochemical markers and histological findings. Reperfusion significantly increased the serum levels of thrombin-antithrombin complex and enhanced PAR1 expression in the intestinal mucosa. The levels of both intraluminal hemoglobin and protein were significantly increased in the I-R group. The mucosal myeloperoxidase activity and expressions and/or productions of cytokine-induced neutrophil chemoattractant-1 and TNF-alpha were significantly increased after I-R. These increases were inhibited by the treatment of rat with antithrombin intravenously before I-R at a dose of 30 U/kg. These results suggest that the thrombin/PAR1 pathway plays an important role in the production of these cytokines during I-R and that antithrombin exerts potent anti-inflammatory effects on this injury via inhibition of proinflammatory cytokines.

Inhibition of coagulation proteases Xa and IIa decreases ischemia–reperfusion injuries in a preclinical renal transplantation model

“…25 Herein, in vivo, we observed a decrease of TAT formation at 60 min post-reperfusion when the kidney was preserved with anticoagulants. Since a direct link between thrombin activity and pro-inflammatory response has been established in IR context, in mice 26 and rat, 27 our results further suggest that EP217609-dependent "thrombin" inhibition may limit inflammation.…”

R-spondin3 prevents mesenteric ischemia/reperfusion-induced tissue damage by tightening endothelium and preventing vascular leakage

“…Several studies have implied that tissue hypoxia affects the integrity of adherens junctions and promotes increased vascular permeability. This breakdown of the barrier has been shown to be associated with increased levels of inflammatory mediators such as thrombin, histamine, and VEGF (27), which further contribute to adverse outcomes of vascular dysfunction. Vascular permeability has thus become a hallmark of numerous autoimmune and inflammatory conditions including I/R (28,29).…”

Proinflammatory role of protease-activated receptor-2 in intestinal ischemia/reperfusion injury in rats

“…it was previously reported that cinc-1 levels increased during small intestinal i/r injury and that cinc-1 was related to the extent of mucosal damage with neutrophil accumulation (21,22,35). in addition, a previous study demonstrated that pre-treatment with an anti-cinc-1 monoclonal antibody attenuated reperfusion injury in the small intestine, in association with a reduction in TnF-α and MPo levels, thereby prolonging survival (36).…”