| Organ transplantation is the treatment of choice for patients with end-stage organ failure. The incidence of acute rejection (AR) of liver allograft is reported to be between 40% and 60%. Therefore, the immune tolerance of liver transplantation remains one of the keys. High throughput genomics and proteomics techniques have facilitated a better understanding of immune tolerance.To observe the acute rejection and the immune tolerance after orthotopic liver transplantation in rats between inbred line Wistar and closed colony SD. We established models with Kamada's two cuff technique and then randomly divided into three groups:Group A:SD→SD;Group B:Wistar→SD; Group C:F protein intervention:intrathymic injection one week before surgery. Three recipients were killed after 3d,6d,9d and 100d. Representative liver allografts tissue and serum specimens were collected for corresponding detection. Remaining recipients were observed for long-term survival. During the experiment 108 models were established, the total success rate was 62.96%(68/108). The success rate was only 24.44%(11/45) during the protophase of model manufacture. Afterward the success rate gradually comes up to more than 90%.The causes of death were hemorrhage, anesthetic accident, long anhepatic phase, thrombus formation and so on. In recipients of A group, rejection was not founded. In recipients of B group, I grade rejection occurred at 3rd day and rejection gradually reached the peak 6 days after transplantation, and most died between 12 to 21 days after operation. In recipients of C group rejection was not founded either. Conclusions:(1) Two cuff technique may provide a practical and stable experimental model.(2)The keys of establishing successful models include high quality donor liver, practiced microsurgical techniques and short anhepatic phase.(3) The model can be used to study acute rejection or immunologic tolerance effectively. (4) F protein can induce immune tolerance in liver transplantation. Application of nanoUPLC-MS/MS technology to research the differentially expressed proteins at 7 days after rat liver orthotopic transplantation between immune tolerance group and syngeneic control group and to have a preliminary study of the immune tolerance. Establish immune tolerance group(Wistar→SD+F protein) n=5, establish syngeneic control group (SD→SD) n=5. After 7 days kill five receptors, obtain liver tissues, extract total protein, after Trypsin hydrolysis, use Waters exclusive patented MSe unmarked quantitative techniques, through the technology of nanoUPLC-MS/MS analyse the peptide hydrolysis, database search identify the protein, bio-informatics tools for the identification of protein functional classification. Comparing the two groups, the two samples simultaneously exist in most of the proteins increases, of which 9 protein spots were significantly up-regulated expression levels, changes in the ratio of 2.59 times. Combination of database search to be identified, these were significantly increased in the main function of the protein molecules with the cytoskeleton, ion binding, oxidative stress response, energy metabolism and other related. This study established a proteomic analysis methods for rat orthotopic liver transplantation with nanoUPLC-MS/MS, carried out a preliminary study for F protein induced immune tolerance after rat liver transplantation, provided quantitative indicators for the establishment of animal models and provided direction and data support for identifying potential biological markers.
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