| Objective: Weestablished the model of fatty liver ischemia reperfusion injury model, studying the effect of resveratrol preconditioning on the expression of Gadd45 a and the possible mechanism of the protective effect of resveratrol on the protective effect of fatty liver on ischemia reperfusion inury. Method: This experiment is divided into two parts. The first part is the pre experimental stage(to establish the model of fatty liver ischemia and reperfusion injury in mice). The second part for the formal test stage, namely in the successful established mice fatty liver ischemia and reperfusion injury model. Objective to study the effects of resveratrol on the pre processing protective effect on fatty liver ischemia reperfusion injury and the possible mechanisms.1.Pre-experimental stage:select 8-12 weeks healthy, male KM mice(experimental animal center of university of south China) 14 day, body weight 17-24 g. Mouse fatty liver model, which is widely recognized in the world, is a model of nonalcoholic fatty liver disease(NASH) induced by methionine and choline. Before the experiment, methionine and choline were given a diet of 14 d, and the experimental day was established with Pringle's s method to create a model of hepatic ischemia reperfusion in mice. The blood vessel was clamped for 20 minutes. Ischemia reperfusion after 2H from mouse liver tissue assay, to observe the pathological changes of liver tissue.2. Formal experimental phase:select 8-12 weeks healthy male KM 21 mice, weighing between 17-24g; were randomly divided into 3 groups, 7 rats in each group: a group of mice were given normal diet 14 d before the experiment; before the experiment 2D and experiment on the same day, every morning to give Sodium Chloride Injection 0.9% intraperitoneal injection 1 times(calculated in accordance with the 30 mg /kg dose group); second mice were given the lack of methionine and choline diet 14 d before the experiment; before the experiment 2D and experiment on the same day, every morning to give Sodium Chloride Injection 0.9% intraperitoneal injection 1 times(calculated 30mg/kg dose); third groups of mice were given the lack of methionine and choline diet 14 d before the experiment, before the experiment 2D and experiment on the same day, every morning to give 10% resveratrol injections 1 times(calculated in accordance with the dose of 30mg/kg). In the first group, the first group was simply switched to the abdomen, without clipping the hepatic pedicle vessels; the second group and the third group of s' Pringle 'method was established to create the mouse fatty liver ischemia reperfusion model, clipping the hepatic pedicle vascular 20 minutes. To be a successful modeling of mice in each group, and clipping in liver tissue of mice and extraction of serum test; mice liver function observed the pathological changes of liver tissue; used in liver tissue was determined with Western blot Gadd45 a expression level. Finally, SPSS17.0 software was used to analyze the data of each group. Result: 1.Pre-experimental stage:a total of 14 mice were selected to establish the model of fatty liver ischemia reperfusion injury. The methods of feeding the mice fatty liver, surgical aseptic technique, surgical open liver pedicle vascular occlusion, anesthetic dose control, surgical fluid replacement and other issues of skills, including 2 cases of death. The cause of death was caused by the injury of the diaphragm during open surgery. The remaining 2 cases were still alive after 12 hours of ischemia and reperfusion. Liver tissue sections of the lesion showed that 12 cases of liver of mice have marked macrovesicular steatosis, lobular multiple with inflammatory cell infiltration, fibrosis and cirrhosis. The successful model of fatty liver ischemia reperfusion injury in mice. 2.Formal experimental phase:(1)No death cases were found in the three groups. After 14 d B, the MCD diet, C group of mice liver cells had bullous steatosis.(2) Liver function: using the high sensitivity of the biochemical analyzer to measure the concentration of serum ALT; one group of mice serum ALT fluctuated within the normal range P<0.05); the second quarter of the three groups of mice serum alanine aminotransferase(ALT), were significantly higher than those of the normal range, and by resveratrol pre treatment of the three groups of mice than a second group of mice serum ALT have varying degrees of decline; statistical software to analyze the results suggest that serum results have obvious differences, <0.01, statistical significance. Resveratrol pretreatment could significantly reduce the degree of hepatic ischemia reperfusion injury and protect the liver function.(3)Pathology of hepatic tissue: the first group mice hepatic lobule and hepatic sinusoid morphology normal, liver there was no inflammatory cell infiltration and necrosis, no macrovesicular steatosis; second, and three groups of mice liver cells have different degrees of bullae fatty degeneration, inflammation invasion and necrosis of liver cells, but after resveratrol pre processing of the third group of fatty liver cell degeneration, necrosis and inflammation were significantly decreased than that of the second group, suggesting that resveratrol can reduce the fatty liver ischemia again reperfusion injury in fatty degeneration and inflammatory necrosis.(4) Western blot: the first group of Gadd45 a expression is very weak; the second group and the third group of Gadd45 a expression level was significantly higher than that of the first group, but after resveratrol pre processing of the third group of Gadd45 a expression levels than those in the second group, by statistical analysis of each group of Gadd45 a expression level was statistically significant(P < 0.01). Resveratrol can increase the expression level of Gadd45 a in hepatic ischemia reperfusion injury. Conclusion: Resveratrol could obviously reduce the degree of hepatic ischemia reperfusion injury in mice, thus protecting liver function. Resveratrol can improve the expression level of Gadd45 a in liver tissue of fatty liver ischemia reperfusion injury. |
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