Role of the thrombin/protease-activated receptor 1 pathway in intestinal ischemia-reperfusion injury in rats.
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.
Tsuboi
Hisato
H
Department of Inflammation and Immunology, Kyoto Prefectural Univ of Medicine, Kawaramachi-Hirokoji, Kyoto, Japan.
Naito
Yuji
Y
Katada
Kazuhiro
K
Takagi
Tomohisa
T
Handa
Osamu
O
Kokura
Satoshi
S
Ichikawa
Hiroshi
H
Yoshida
Norimasa
N
Tsukada
Minoru
M
Yoshikawa
Toshikazu
T
D000818:Animals; D000990:Antithrombin III; D000991:Antithrombins / Q000494:pharmacology; D054360:Chemokine CXCL1; D019743:Chemokines, CXC / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression / Q000187:drug effects; D006454:Hemoglobins / Q000032:analysis; D007082:Ileum / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D007413:Intestinal Mucosa / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism; D007421:Intestine, Small / Q000187:drug effects / Q000473:pathology / Q000503:physiopathology; D007422:Intestines / Q000187:drug effects / Q000473:pathology / Q000503:physiopathology*; D008297:Male; D010447:Peptide Hydrolases / Q000097:blood; D009195:Peroxidase / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D044463:Receptor, PAR-1 / Q000502:physiology*; D015427:Reperfusion Injury / Q000473:pathology / Q000503:physiopathology* / Q000517:prevention & control; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology; D013917:Thrombin / Q000502:physiology*; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics / Q000378:metabolism
D000991:Antithrombins; D054360:Chemokine CXCL1; D019743:Chemokines, CXC; C513297:Cxcl1 protein, rat; D006454:Hemoglobins; D012333:RNA, Messenger; D044463:Receptor, PAR-1; D014409:Tumor Necrosis Factor-alpha; C046193:antithrombin III-protease complex; D000990:Antithrombin III; D009195:Peroxidase; D010447:Peptide Hydrolases; D013917:Thrombin