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《World Chinese Journal of Digestology》 2005-05
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Effect of endotoxemia on enterocyte apoptosis and PCNA expression in baby rats

Xiu-Qing Wu, Hong Wang, Mei Sun, Department of Pediatric, the Second Affiliated Hospital of China Medical University, Shenyang 110004, Liaoning Province, China Qing-Jie Lv, Department of Pathology, the Second Affiliated Hospital of China Medical University, Shenyang 110004, Liaoning Province, China Zhuo Zhou, Central Laboratory, the Second Affiliated Hospital of China Medical University, Shenyang 110004, Liaoning Province, China  
AIM: To investigate the effect of endotoxemia on enterocyte apoptosis and proliferation in baby rats, to provide theoretical evidence for anti-apoptotic or growth-promoting therapy. METHODS: The model of endotoxemia of baby rats was established by intraperitoneal injection of either saline (control) or endotoxin (4 mg/kg of Escherichia coli lipopolysaccharide). 4 cm lower ileum was collected at 0 h (control group only), or within 10 min after dead (endotoxin group only), 2,4,6,24, and 72 h after injection respectively. The pathologic changes of small intestine villus were observed by hematoxylin-eosin (HE) staining. Apoptotic cells were detected by terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) staining and proliferating cell nuclear antigen (PCNA) expression was measured by immunohistochemistry staining. RESULTS: Apoptotic activity in each endotoxin group was significantly higher than that of control group (37.4 ± 7.1, 44.1 ± 9.3,46.6 ± 8.2, 54.7 ± 9.0,45.3 ± 7.2 and 33.9 ± 6.1 vs 0.02 ± 0.01, P0.01 respectively). PCNA expression in each endotoxin group was significantly lower than that of corresponding control group (P0.05). The markedly increased apoptosis index started from 2h after injection of endotoxin, and increased continually there after, peaked at 24 h, retrieved to the level of 6 h at 72 h. The significantly decreased PCNA expression was found from 2 h (30.2 ± 8.3 vs 74.6 ±16.2, t= 6.874, P0.01) after injection of endotoxin, reached the lowest level at 4h (21.4 ± 6.7 vs 74.6 ± 18.7, t = 7.566, P0.01) and 6h (23.9 ± 14.7 vs 73.6 ± 13.7, t=6.999, P0.01), retrieved to the level of 2 h at 24 h (35.8 ± 11.4 vs 76.3 ± 11.8, t = 7.010, P0.01), and did not get normal at 72 h (59.6 ± 18.0 vs 78.7 ± 16.9, t = 2.185, P= 0.046). Apoptosis index (33.9 ± 6.1) and PCNA expression (20.2 ± 9.2) in dead group were close to those of 2 h and 4 h after injection of endotoxin respectively. Comparison was made by the time points between ratios of PCNA expression and apoptosis index in the same hours. It was found that the ratios of endotoxin group (0.8 ± 0.3, 0.5 ± 0.1, 0.5 ± 0.3,0.6 ± 0.2,1.3 ± 0.3, and 0.6 ± 0.2 respectively) were significantly lower than that of control group [(3.2 ± 0.8) × 103-(3.7 ± 0.7) × 103] (P0.01), and that the ratio of dead group was close to those of 4, 6, 24 h after injection of endotoxin group, and it was approximately the lowest level. CONCLUSION: Enterocyte apoptotic activity is increased and PCNA expression is decreased during endotoxemia in baby rats, and the imbalance between apoptosis and proliferation is one of the pathologic mechanisms of gut barrier impairment during severe infection.
【Fund】: 教育部留学回国人员科研启动基金资助项目.No.[2001]345~~
【CateGory Index】: R574
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