| PrefaceLiver transplantation has become an important and effective method for treating terminal liver diseases and acute liver disfunction. Whereas ischemia -reperfusion injury (I/R I) , whose mechanism is not completely demonstrated, is one of the main causes responsible for liver transplantation failure. The ischemia — reperfusion injury was regarded as a physical reaction extend all over the body. Melatonin ( Mel) is a kind of nerve incretion secreted by pineal gland cells. It's becoming one of the research hotspots owning to the complex physiological and pharmacological effects on multiple systems and organs. Domestic reports about Mel on liver I/R I are rare. In this study, in order to clarify the influence and mechanism of Mel on I/R I to kidney in hepatic ischemia reperfusion injury in rats, I/R I models were set up, samples according to different time after reperfusion were collected, biochemistry series ( ALT, AST, AKP, BUN, Cr, UA ) in serum, anti - oxidase series ( SOD, GSH. px) and terminal productions of lipid peroxidation ( MDA) in liver and kidney tissue were determined. Meanwhile, hematoxylin - eosin (H. E) and immunohistochemical staining were employed to observe the morphologic change under light microscope as well as liver and kidney cells under electron microscope. Mechanism of the protective effects of Mel on liver I/R I to kidney and liver function was discussed in sub - cellular level. and evidence the phase of protective effects of Mel to kidney comparing with to liver.Material150 male Wistar rats weighing 190 -210 grams, aging 6 -7weeks were di-vided into three groups at random; Mel exposure group, alcohol solvent control, group and saline control group. According to Kobayashi, we blocked up the left branches of portal vein, hepatic artery for 60 min and then opened them to establish liver I/R I models in rats. In each group, samples were collected in 0. 5h, lh, 6h, 12h, 24h after reperfusion respectively. One gram of Mel was purchased from Sigma company and was stored in low temperature after being dissolved by alcohol and saline. 20mg/kg of Mel was injected IP in rats 30 min before experimentation. The duplicate concentration of alcohol and the same volume of saline were injected in control groups as substitution.MethodMeasurement of hepatic enzyme series ( ALT, AST, AKP, BUN, Cr, UA ) in serum by auto biochemical analyzer.Measurement of anti — oxidase series (SOD, GSH. px) and terminal productions of lipid peroxidationin (MDA) in liver and kidney tissue.Pathological examination of I/R I liver and kidney tissue.Immunohistochemical staining of ICAM - 1.Observation of hepatic and kidney cells under electron microscope.Statistics: Data was expressed as X ± S. Statistics analysis was performed with One - Way analysis of variation ( ANOVA) and SNK test with the help of software SPSS 13.0Result1. Biochemistry series in serum in rats:1. IThe level of ALT measured in various time after reperfusion in Mel group was totally significantly lower than that in alcohol and saline control groups corresponding ( p < 0.05).1.2The level of AST measured in 6h and 12h after reperfusion in Mel group was significantly lower than that in alcohol and saline control groups correspond-ing( p<0.05).1. 3The level of AKP measured in 24h after reperfusion in Mel group was significantly lower than that in alcohol and saline control groups corresponding ( p<0.05).1. 4The level of BUN measured in 24h after reperfusion in Mel group was significantly lower than that in alcohol and saline control groups corresponding ( p <0.05). The level of BUN measured in 24h in every groups was significantly higher than that in other period of time.1.5The level of Cr measured in 24h after reperfusion in Mel group was significantly lower than that in alcohol and saline control groups corresponding ( p <0. 05). The level of Cr measured in 24h in every groups was significantly higher than that in other period of time.1. 6The difference of UA measured in every period of time and in every groups was not significant.The difference of the above six serum biochemistry indexes corresponding between the two control groups was not significant.2. Measurement of anti — oxidase series and terminal productions of lipid peroxidationin in liver and kidney tissue.2. 1 SOD, GSH. px, MDA in liver tissueThe level of SOD measured in 12h, 24h after reperfusion in Mel group was significantly higher than that in alcohol and saline control groups corresponding ( p<0.05).The level of GSH. px measured in 12h, 24h after reperfusion in Mel group was significantly higher than that in alcohol and saline control groups corre-sponding( p <0. 05 ) .The level of MDA measured in 6h,12h, 24h after reperfusion in Mel group was significantly higher than that in alcohol and saline control groups corre-sponding( p <0.05).The difference of the above three variables corresponding between the two control groups was not significant.2.2 SOD, GSH. px, MDA in kidney tissueThe level of SOD measured in 24h after reperfusion in Mel group was significantly higher than that in alcohol and saline control groups corresponding ( p <0.05). The level of SOD measured in 24 h in every groups was significantly higher than that in other period of time.The level of GSH. px measured in 24h after reperfusion in Mel group was significantly higher than that in alcohol and saline control groups corresponding ( p <0.05). The level of GSH. px measured in 24h in every groups was significantly higher than that in other period of time.The level of MDA measured in 12h, 24h after reperfusion in Mel group was significantly higher than that in alcohol and saline control groups corresponding ( p <0.05) . The level of MDA measured in 24h in every groups was significantly higher than that in other period of time.The difference of the above three variables corresponding between the two control groups was not significant.3. Immunohistochemical staining of ICAM - 1 of liver tissue in rats.3.1 liver tissueThe express level of ICAM - 1 (% ) measured in various time after reperfusion in Mel group was totally significantly lower than that in alcohol and saline control groups corresponding (p < 0. 05 ) .3. 2 kidney tissueThe express level of ICAM - 1 ( % ) measured in 12h, 24h after reperfusion in Mel group was totally significantly lower than that in alcohol and saline control groups corresponding (p < 0.05).4. H. E straining of liver tissue in rats (see the following pictures).5. Observation of liver tissue under electron microscope in rats (see the following pictures).DiscussionThe change of the biochemistry series indexes (ALT, AST, AKP,BUN, Cr,UA) after reperfusion showed lessened damage in Mel group in comparison with that in the two control groups, which demonstrated exotic Mel improved the hepatic and kidney function after reperfusion, and the phase of protection to kidney was postponed comparing with to liver. That postponement may be caused of...
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