Protection Of Resveratrol On Mice Hepatic Ischemia-reperfusion Injury

Background and Objective: Liver transplantation has become the most effective choice for the treatment of end-stage liver disease and acute liver failure.The liver and biliary tract after transplantation with normal biological function evaluation is the key to successful operation. Cold and warm ischemia reperfusion injury(ischemia-reperfusion, injury, IRI) are the main cause for liver without function after transplantation. While the drug pretreatment can reduce the liver, bile duct transplantation ischemia reperfusion injury, and may reduce the occurrence of hepatic dysfunction. Resveratrol is a kind of polyphenol compound and has a strong biological activity with antioxidant, anti-inflammatory, anti thrombosis, anti- aging, immune regulation and anti tumor effect, to prevent cardiovascular disease and cancer.It can be extracted from grapes, Polygonum cuspidatum, peanut, mulberry and other plants. At present, resveratrol is mainly used for the protection of organs such as heart, brain, kidney andso on, however the protection of it on liver and bile duct ischemia reperfusion injury is seldom studied.In this experiment,We hope to establish a stable liver and biliary tract ischemia-reperfusion injury model in mice through the simulation of liver transplantation for warm and cold ischemia to reperfusion rewarming. Then we use the model to study the protection of resveratrol pretreatment on hepatic and extrahepatic bile duct ischemia reperfusion and its possible mechanism.This will be beneficial for the safe and rational use of resveratrol and provide a theoretical basis for clinical application.Martialand and Methods: This experiment is divided into 2 stages, in the first phase we explored and established a liver ischemia-reperfusion model in mice.In the second stage, after the success of the model, we studied the protective effect of resveratrol over the long period of the administration of the liver and bile duct suffering from ischemia reperfusion injury.The first stage:30 KM mice were deployed to establish the ischemia reperfusion injury model.We referred and improved Zhao Hongfeng’s orthotopic liver transplantation model in rats.It used 4 containing 12.5 ℃U/ml heparin ringer’s lactate solution to perfuse into the superior mesenteric vein,abdomina laort and extrahepatic biliary with constant pressure,and then sutured blood vessels with “trestle method”.The operation time, the success rate and the effect of cold perfusion were obs-erved and recorded.The second stage: Healthy, adult, clean KM mice 60 were randomly divided into three groups.Group A were as the blank control groups,Group B were as ischemia reperfusion groups, Group C were as ischemia reperfusion +Resveratrol pretreatment groups.The mice in Group C were lavaged with resveratrol(20mg/kg per mice)2 week before the experiment.The operations of mice in Group A were just abdomen open and closed.Both Group B and C established the model according to the first part experiment. After 2 hours of Ischemia reperfusion we obtained samples from mice.The postoperative animal survival rates and liver function were tested and calculated in mice. The pathological changes in liver and common bile duct were observed by electron microscope. The expression of Gadd45 a was measured by Western-blot.Results:(1)The first stage:the sueeess rate after the operation was 66.7%(20/30).hot and cold ischemia and reperfusion time could be controlled accurately. Anhepatic phase was controlled within 20 minutes.The effect of double constant pressure perfusion with biliary flushing is fine.we observed the bile duct wall capillary that could not almost contain red blood cells.The model reached the requirement of liver warm and cold ischemia reperfusion injury,and it had the features of simple operation, strong stability, relatively short operation time and strong specificity.(2)The second stage:1. Group A had no casualties, Group B and C bothdied of 5 mice, Group B and C had not statistical difference(P > 0.05) on suceess rate. 2. Serum ALT level: Serum ALT levels of Group A after postoperative were in normal range;Compared to Group A,Group B and C significantly increased the levels of serum ALT in the same time point, but the levels of Group B were lower than that of group B, they existed statistical significance(P < 0.05), It suggested liver damage of Group C was more lighter than Group B. 3. Liver and extrahepatic biliary pathologic change:The hepatic cells and tissue structures in group A were normal.The pathological changes in group B were as follows:hepatic cells were obviously oedematouse and appeared vacuoles degeneration;their cytoplasm were dyed lightly;liver blood sinus almost disappeared;common bile duct epithelial cells falled off and their epithelial structure were significantly incomplete.However, compared to group B, hepatic cells in Group C were mild swelling and had no vacuoles degeneration.The structural damage of liver tissue and common bile duct in Group C was significantly reduced. 4. Western blot : The expression of Gadd45 a in Group A was weak positive.The expression of Gadd45 a in Group C was higher than that in Group B(P<0.01).Con Clusion:1.Resveratrol can reduce blood serum aminotrans-ferase levels of mice hepatic caused by warm and cold ischemia-reperfusion injury and mitigate the liver and extrahepatic biliary pathological damage.2. Resveratrol can promote early expression of Gadd45 a in the liver tiss-ue of ischemia reperfusion.