Study Of The Mechanism Of All-trans Retinoic Acid In Liver Ischemia And Reperfusion Injury

Backgroud:Liver ischemia and reperfusion injury is a frequently encountered problem in clinical work. With the state canceled cadaveric liver donor transplant, liver donation after circulation death has become the main source of liver transplantation. The degree of ischemia and reperfusion injury of liver donation after circulation death, however, will obviously affect the recovery of graft function, and even determine the success or failure of liver transplant. Therefore, how to alleviate the degree of liver ischemia and reperfusion injury in the process of liver donation after circulation death will be the key to the success of liver transplantation. All-trans retinoic acid is the commonly used drug for the treatment of acute myeloid leukemia in clinic. It has been found that all-trans retinoic acid also reduce liver ischemia and reperfusion injury, but the mechanism is unclear. Hence, the purpose of this experiment is to uncover the specific mechanism of all-trans retinoic acid in liver ischemia and reperfusion injury.Method:In vivo, the 70% hepatic ischemia model was applied in the experiment, and it included Sham group, ischemia and reperfusion injury group and all-trans retinoic acid given group. The mice were sacrificed after blood collection at the reperfusion time. Then, blood sample was used to test liver function, and the other tissue sample was detected by western blotting and reverse transcription-polymerase chain reaction. In vitro, Hydrogen peroxide and lipopolysaccharides were respectively used to stimulate FL83B and RAW 264.7. Experimental groups included Sham, hydrogen peroxide or lipopolysaccharides, all-trans retinoic acid pretreatment and LE540 pretreatment group. Cells were collected at the corresponding time point, and then were analysed by western blotting and reverse transcription-polymerase chain reaction.Result:Firstly, liver function and its pathology score in IR injury group were higher than in Sham group,and there was statistical difference; Secondly, comparing with IR injury group, liver function and liver histopathological score were lower, and the levels of inflammatory cytokines tumor necrosis factor α、 interleukin-6 and interleukin-1β also had a significant decline in ATRA pretreatment group; Thirdly, in ATRA pretreatment group, the levels of retinoic acid receptor α、Bcl-2、p-Akt、 Beclinl、LC3Ⅱ/LC3 Ⅰ protein were upregulation than the IR injury group, however, the levels of cleaved-caspase 3、Foxo1、TLR4、NF-κB p65 and p62 were downregulation in comparison with IR injury group; Fourthly, ATRA pretreatment with FL83B in comparison with hydrogen peroxide treatment, decreased the apotosis and the expression of cleaved-caspase 3、Foxo1、p62, and increased the expression of retinoic acid receptor α、Bcl-2、p-Akt、 Beclinl、LC3 Ⅱ/Ⅰ protein, LE540 pretreatment exacerbated the damage in comparison with hydrogen peroxide group; Sixthly, in ATRA pretreatment group, the levels of inflammatory cytokines tumor necrosis factor α interleukin-6 and interleukin-1β were less than in LPS group, the levels of p62, Foxo1, TLR4 and NF-κB p65 also had similar trends, but the levels of retinoic acid receptor a, Beclinl, LC3Ⅱ/Ⅰ and p-Akt were higher, in contrast, LE540 had an opposition role in comparison with ATRA.Conclusion:Firstly,70% hepatic ischemia and reperfusion injury model was establishedsuccessfully;Secondly, ATRA can alleviate the liver ischemia and reperfusion injury by decreasing liver cells apotosis and regulatinginnate immunity response;Thirdly, the protective role of ATRA on autophagy maybe related to activate retinoic acid receptor a.