Experimental: ischemia-reperfusion injuryActivation of immediate early genes in relation to proliferation and apoptosis of enterocytes after ischemia-reperfusion injury of small intestine
Section snippets
Materials and methods
Whole small intestines of male Lewis rats were isolated and clamped for 30 minutes, and then reperfused (n = 6). Tissue specimens were taken at 0, 15, 30, 60, 90, 120, 180, 240, and 360 minutes after reperfusion. Total RNA was extracted from mucosa, and expression of c-fos and c-jun were quantified by semi-quantitative RT-PCR method. Proliferation of the epithelial cell was visualized by immunohistochemisty for proliferating cell nuclear antigen (PCNA), and apoptosis was detected by in situ
Results
The c-fos expression was activated transiently, peaked at 30 minutes after reperfusion, and immediately declined. Activation of c-jun was slower, peaking at 90 minutes after reperfusion, with an expression prolonged more than c-fos. The number of PCNA-positive cells increased at 60 to 360 minutes after reperfusion, and TUNEL-positive cells increased later than PCNA-positive cells, at 120 to 360 minutes after reperfusion. The relationship between the expression of m-RNA and the apoptosis and
Conclusion
Patterns of mRNA expression of c-fos and c-jun in rat intestine after ischemia-reperfusion were similar to those in mouse liver.2 After I/R stress, transcriptional activation of c-fos and c-jun was observed in the small intestinal epithelium. Sequential expression patterns of those genes were believed to be related to the cellular responses such as proliferation and apoptosis. Increase of c-jun expression correlated with increase of TUNEL-positive cells, therefore c-jun was suggested to be one
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This work was partly supported by Grant-in-Aid for Scientific Research (B2) from the Ministry of Education and Science, Japan.