| Objective: Hepatic ischemia/reperfusion injury (IRI) is an important nonimmunologic factor and remains a significant problem and limitation of liver transplantation and may result in liver failure, remote organ failure, and even death. For this reason, the protection of liver against ischemia/reperfusion injury remains one of the major nonimmunologic problems of liver transplantation.In addition, hepatic ischemia/reperfusion injury may increase the expression of some antigens associated with allograft rejection, such as MHC class I and MHC class II molecules, and thus make the allografts more " immunogenic" and therefore more susceptible to rejection and dysfunction. Increaseing evidences demonstrate that MHC class I chain-related antigen A (MICA) and B (MICB) may play a role in allograft rejection, however, whether IRI affects the expression of MIC has not been documented.Our previous studies demonstrate that Yisheng injection (YS), a herbal preparation developed from traditional Chinese medicine, can attenuate ischemia/reperfusion injury in vitro and in vivo. Based on the previous results, the present study explored the effect and mechanism of YS on hepatic ischemia/reperfusion injury; determined the effect of IRI on the expression of RAE-1 (retinoic acid early inducible) and H60 genes, which are the functional homologs of human MIC in mouse; and examinated the effect of YS intervention on the expression of RAE-1 and H60 genes.Methods: Mice partial hepatic warm ischemia/reperfusion model was used in this study.1. C57BL/6 mice were randomly assigned into five experimental groups as follows: (1) sham operation group, (2) I/R group, (3) small-dose (5mg/kg of YS) treated group, (4) middle-dose (10mg/kg of YS) treated group, (5) large-dose (20mg/kg of YS) treated group.YS with different doses (5, 10, 20 mg/kg) was injected intraperitoneally 24 h and 1 h before ischemia and a third dose was injected intravenously just before reperfusion. The sham and I/R group received same volume of sterile saline. After 90 min of ischemia and 6 h of reperfusion mice were sacrificed and liver tissue (from the ischemic lob) and blood samples were taken for analysis. The ALT, AST and LDH levels in the sera were measured, and the levels of SOD, MDA, MPO and edema of ischemic liver were examinated. The serum level of TNF- a was assayed by enzyme-linked immunosorbent assay (ELISA), and the expression of TNF-a and ICAM-1 was determined by immunohistochemical assay and reverse transcription polymerase chain reaction (RT-PCR).2. BALB/C mice were randomly assigned into sham and I/R groups. After90 min of ischemia the left lob of the liver in the I/R group was reperfused. Mice were sacrificed at intervals of 1, 2, 3, 5, 7, 10, 15, 20, and 30 days postoperatively, and the normal, sham and ischemic liver tissues were collected. The expression of RAE-1 and H60 genes was measured by RT-PCR and real-time quantitative PCR.3. BALB/C mice were randomly assigned into three groups as follows: (1) sham operation group, (2) I/R group, (3) YS (20mg/kg) treated group. After 90 min of ischemia the left lob of the liver in the I/R and YS treated groups was reperfused. YS (20 mg/kg) was injected intraperitoneally 24 h and 1 h before ischemia and a third dose was injected intravenously just before reperfusion. After operation YS treated group was given YS (20mg/kg/d) for 7 days. The sham and I/R group received same volume of sterile saline. Mice were sacrificed at 7 days after operation, and the normal, sham, ischemic and YS treated liver tissues were collected. The expression of RAE-1 and H60 genes was measured by RT-PCR and real-time quantitative PCR.Results: (1) YS treatment can effectively reduce the hige levels of ALT, AST, LDH and TNF-a in the sera caused by 90 min hepatic warm ischemia and 6 h of reperfusion. After YS treatment, the morphology and structure of vascular endothelium are relatively normal and the lymphocyte infiltration reduced.(2) YS treatment can effectively reduce the oxidative stress by elevating the SOD level and reducing the MDA lev... |
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