Altered distribution of tight junction proteins after intestinal ischaemia/reperfusion injury in rats.
Tight junction (TJ) disruptions have been demonstrated both in vitro and more recently in vivo in infection. However, the molecular basis for changes of TJ during ischaemia-reperfusion (I/R) injury is poorly understood. In the present study, intestinal damage was induced by I/R in an animal model. As assessed by TUNEL and propidium iodide uptake, we showed that I/R injury induced apoptosis as well as necrosis in rat colon, and the frequency of apoptotic and necrotic cells reached the maximum at 5 hrs of reperfusion. Immunofluorescence microscopy revealed that claudins 1, 3 and 5 are strongly expressed in the surface epithelial cells of the colon; however, labelling of all three proteins was present diffusely within cells and no longer focused at the lateral cell boundaries after I/R. Using Western blot analysis, we found that distribution of TJ proteins in membrane microdomains of TJ was markedly affected in I/R injury rats. Occludin, ZO-1, claudin-1 and claudin-3 were completely displaced from TX-100 insoluble fractions to TX-100 soluble fractions, and claudin-5 was partly displaced. The distribution of lipid raft marker protein caveolin-1 was also changed after I/R. I/R injury results in the disruption of TJs, which characterized by relocalization of the claudins 1, 3 and 5 and an increase in intestinal permeability using molecular tracer measurement. I/R injury altered distribution of TJ proteins in vivo that was associated with functional TJ deficiencies.
Li
Qiurong
Q
Institute of General Surgery, Jinling Hospital, Nanjing, China. liqiurong@yahoo.com
Zhang
Qiang
Q
Wang
Chenyang
C
Liu
Xiaoxiang
X
Qu
Linlin
L
Gu
Lili
L
Li
Ning
N
Li
Jieshou
J
D000818:Animals; D062445:Claudin-1; D062465:Claudin-3; D062507:Claudin-5; D057167:Claudins / Q000096:biosynthesis; D003902:Detergents / Q000494:pharmacology; D007413:Intestinal Mucosa / Q000378:metabolism; D007422:Intestines / Q000473:pathology*; D008297:Male; D021962:Membrane Microdomains / Q000378:metabolism; D008565:Membrane Proteins / Q000096:biosynthesis; D018613:Microscopy, Confocal / Q000379:methods; D046529:Microscopy, Electron, Transmission / Q000379:methods; D011092:Polyethylene Glycols / Q000494:pharmacology; D051381:Rats; D017208:Rats, Wistar; D015427:Reperfusion Injury / Q000473:pathology*; D019108:Tight Junctions / Q000473:pathology*
D062445:Claudin-1; D062465:Claudin-3; D062507:Claudin-5; D057167:Claudins; C568114:Cldn1 protein, rat; C568124:Cldn3 protein, rat; C491510:Cldn5 protein, rat; D003902:Detergents; D008565:Membrane Proteins; C551282:polyethylene glycol monooctylphenyl ether; D011092:Polyethylene Glycols