Effect Of Combination Of Creatine Phosphate Sodium, Ischemic Preconditioning And Ischemic Postconditioning On The Intestinal Ischemia-reperfusion Injury In Rats

Objective: To investigate the effect of combination of creatine phosphate sodium treatment, ischemic preconditioning and ischemic postconditioning on the intestinal ischemia-reperfusion(I/R) injury in rats and its mechanism.Methods: Forty male Wistar rats weighing 250～300g were randomly allocated into 5 groups(n=5):groupⅠsham operation(S);groupⅡI/R; groupⅢcreatine phosphate sodium+I/R+creatine phosphate sodium(C); groupⅣischemic preconditioning + I/R + ischemic postconditioning(IP+IPo); groupⅤcreatine phosphate sodium+ischemic preconditioning+I/R+ ischemic postconditioning(C+IP+IPo ). Intestines were exposed through midline incision and superior mesenteric artery(SMA) were occluded 60min with atraumatic mini-clamp then unclamped for 120min .The intestines turned kermesinus.In S group the intestines were exposed but their SMA were not clamped. In C group creatine phosphate sodium 1mg/kg was given i.v.5min before ischemia and 5min before reperfusion. In IP+IPO group, the superior mesenteric artery were induced by 3 circles of ischemia(5 min) separated by reperfusion(5 min) before ischemia, and 60min ischemia was followed by three 30s episodes of ischemia at 30s intervals for reperfusion. In C+IP+IPo group creatine phosphate sodium 1mg/kg was given i.v.5min before ischemia and 5min before reperfusion, the other operation same as IP+IPO group.The animals were killed at 120min of reperfusion .Blood samples were collected from heart for determination of SOD activity and MDA level concentrations. Intestine specimens were collected immediately for determination of SOD activity and MDA level and water content ,the expression of Bcl-2,Bax and Caspase-3 protein by immuno–histochemistry assay.Results: After I/R, the MDA contents, the PI of Bax,Caspase-3, protein contents and water contents were all significantly increased while the SOD activity, the PI of Bcl-2 were significantly decreased as compaired to the S group(P<0.05).The MDA contents, the PI of Bax,Caspase-3, protein contents and water contents were all significantly lower while the SOD activity, the PI of Bcl-2 were significantly higher in C group, IP+IPo group and C+IP+IPo group as compaired with I/R group(P<0.05).The histologic damage induced by I/R were significantly ameliorated.Compaired with C+IP+IPo group, the SOD activity were significantly lower in IP+IPo group while the PI of Bax ,the MDA contents significantly increased ;the PI of Caspase-3, were all significantly increased in C group .There no significant difference in the rest index among the C group, IP+IPo group, C+IP+IPo group.Cinclusion: C,IP+IPo and C+IP+IPo group can reduce the risk of intestinal dysfunction and injury associated with intestinal I/R injury by reducing apoptosis through enchancement SOD activity, increasing the expression of Bcl-2 protein ,decreasing the Bax and Caspase-3 protein and regulating the Bcl-2/Bax ratio accordingly.C,IP+IPo and C+IP+IPo group can protect the intestine against ischemia-reperfusion injury in rats and C+IP+IPo group is more effective.