| Objective To provide experiment evidence for treating the intestine damage induced by ischemia-reperfusion in intestinal surgical operation and to investigate the protection of puerarin on the model of intestinal ischemia-reperfusion injury in rats. Methods Intestine ischemia-reperfusion injury models were established in SD rats. According to different ischemic and reperfusive time, animals were divided into ischemia-reperfusion groups and in puerarin treating groups. The reperfusion was performed 40 minutes after ischemia in the intestines of the SD rats. The intestine tissues of the experimental groups were taken at Omin , 30 min, 1h, 2h, 3h, 6h, 12h, 24h, 48h, 72h and 96h respectively after the reperfusion and the control groups were treated with pseudo-operation. The mucosa epithelial cells apoptosis and proliferation induced by ischemia-reperfusion were dynamically determined by TUNEL and electron microscope, and immunohistochemistry to detect HSP70, VEGF, PCNA. At the same time, the protection of puerarin on the model of intestinal ischemia-reperfusion injury was observed.Results 1 .The rat intestines were undergone 4 stage changes as incubation, acute, alternative and recovery stages from 40 minutes after ischemia to 96 hours after reperfusion. The epithelial cells apoptosis and histopathologic changes induced by ischemia-reperfusion began at the incubation stage, aggravated at the acute stage. 2.The expression of HSP70 was enhanced in both I/R groups and the puerarin treating group. While the expression of HSP70 in puerarin treating groups were more powerful. The difference of expression of HSP70 between the two groups wassignificant at the same time point (P<0.05). 3.There were both expression of VEGF in endothelial cells of vessels and capillary vessels in basal membrane in two groups after intestine ischemia-reperfusion. The expression of VEGF in puerarin treating group was weaker than that of I/R groups. And the difference of the expression of VEGF between the two groups was significant at the same time point (P<0.05). 4. The expression of PCNA was enhanced in both I/R groups and the puerarin treating group compared to control group. While the expression of PCNA in puerarin treating groups were more powerful in 3 hours after ischemia-reperfusion and lower in 6~48 hours after ischemia-reperfusion. The difference of the expression of HSP70 between the two groups was significant at the same time point (P<0.05). 5. Effect of puerarin on the intestinal ischemia-reperfusion injury: the mucosal histopathologic damages were alleviative in all groups of peurarin treating groups than that of ischemia-reperfusion. So did the apoptosis cells and the expression of VEGF. But the expression of HSP70 was more than that of I/R groups.The expression of PCNA was increased in incubation and acute stages.Conclusios Lit induced an obviously epithelial cells apoptosis and histopathologic changes after a short time ischemia and reperfusion. Most cells turned into convalescence after a short pre-apoptosis reaction, then the intestine mucosal structure and function were repaired. 2. HSP70, VEGF, PCNA contributed to intestine ischemia-reperfusion, and could be one of valued index to the dynamic alteration of intestine ischemia-reperfusion. 3. Puerarin could protect the mucose injury from the intestine ischemia-reperfusion in rats was dued to inhibit the apoptosis during the process of intestine ischemia-reperfusion injury. |
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