| Objective: This experiment by creating the establishment of (IR) animalmodel, to test whether intravenous hydrogen saturated salineischemia-reperfusion injury has a protective effect of large dept., Observethe pathological changes, and from the bile duct epithelial cells Bcl-2/Fas gene expression and apoptosis, etc to investigate hydrogen and itspossible mechanism of saturated saline solution.Methods:32male SD rats were randomly divided into four groups:control group(SO), ischemia reperfusion group (IR), ischemia reperfusion+saline group (IR+NS)group, ischemia reperfusion group with thehydrogen-rich water group (IR+HS), every eight. with1%pentobarbitalsodium (0.6ml/100g) intravenous injection, after anesthesia satisfaction,The rat abdomen with iodine disinfection, Open the abdomen the midlineby the rat’s abdomen,find the common bile duct and duodenum, and thejunctions between the duodenum topical epidural catheters retrogradecatheter in bile total tube, fixed and drainage of bile; Separation ofhepatic portal vein, using noninvasive vascular clamp clip liver left, themiddle of the blood vessels, including arteries, veins and bile duct),blocking blood flow, and keep the unobstructed, blood flow to liver caudate lobe, right time withdraw clip by the regulation, restore bloodsupply, thus made70%of liver ischemia-reperfusion injury model.Groups of SD rats were in ischemia60min, by tail vein injectionhydrogen saturation physiological saline, reperfusion after120minunified left liver tissue and collect bile (both for the two groups beforeand after I/R). Liver tissue were fixed with formaldehyde and frozen-80℃refrigerator, after HE staining pathology examination, to observe theliver tissue pathological morphological changes; immunohisto-chemistry (PV) was observed in the liver tissue of Bcl-2/Fas proteinexpression in intrahepatic bile duct epithelial cells; Tunel method todetect apoptosis; Spectrophotometric method to detect malondialdehyde(MDA) and glutathione (GSH) content,superoxide dismutase (SOD)and catalase (CAT) activity. Above all of the data collected by mean±standard deviation (X±s) said, using Spss17.0statistical software forstatistical processing; The comparison of the sample mean varianceanalysis; Comparing two SNK method. Statistical test (P <0.05) wasconsidered statistically significant difference. Results: the control group(SO group), ischemia reperfusion group (IR), ischemia reperfusion+saline group (IR)+NS group compared with postoperative ischemiareperfusion group with the+hydrogen-rich water+HS group (IR) inliver tissue pathology situation improved; The content of glucose (Glu) inbile rise before and after the magnitude of the decrease in IR values were greater than IR before after (IR); Gamma valley acyl transferase (GGT)in bile before and after the activity in the IR increased amplitude decreaseobviously after (IR values were greater than before the IR value);+IRgroup, the IR+NS group, IR HS group of animals in the modeling ofpostoperative liver tissue MDA content increased significantly, reduceGSH content, SOD activity and CAT activity decreased; Compared withthe IR+NS group, IR+HS group less postoperative liver tissue MDAcontent increased (P<0.01), GSH content decreased less (P<0.01), SODactivity and CAT activity decreased degree also lower (P <0.01).Immunohistochemical method detect the Bcl-2gene and the expressionof Fas gene, IR+HS group of intrahepatic bile duct epithelial cellsNumber of positive cells than IR+NS group. Tunel detection ofintrahepatic bile duct epithelial cells apoptosis, IR and intrahepatic bileduct epithelial cells apoptosis+HS group was obviously lighter than IR+NS group. Conclusion: hydrogen Saturated saline can reduce the injury ofbile duct in liver after hepatic ischemia-reperfusion injury, can inhibitoxidative which is caused by hepatic ischemia-reperfusioninjury, attenuate oxidative damage, can inhibit expressed in bile ductepithelial cells in Bcl-2/Fas protein in liver ofinhibition after hepaticischemia-reperfusion injury, reduce cell apoptosis. |
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