| Objective:(1) To establish a stable liver transplantation (ROLT) model ofEchinococcus multilocularis (E.m) infected rat. Observe survival time, liver function,pathological analysis of transplantated liver between two groups. And explore therelationship between immune of larvae infection and liver transplant immunity.(2) Detectthe expression of serumal IL-17cytokine, IL-17mRNA and IL-17cytokine intransplanted liver. Initially prove expression and its significance of Th17-type cytokinesin E.m infection rats were underwent liver transplantation. Methods:(1) Finished the livertransplantation (ROLT) model using two-cuff technique with Lewis (LEW) rats andBrown Norway (BN) rats. The former were used as donors and BN rats were used asrecipients. BN recipients of experimental group (E.M~+-ROLT) were subcutaneouslyinoculated with Echinococcus multilocularis while the control group (E.M~--ROLT) BN ratsweren’t subcutaneously inoculated with E.m. Observe receptors`survival time of twogroups after liver transplantation.(2) Kill14receptors randomly at7days after surgery.Get5mL blood form inferior vena cava venous and take two portion liver tissue, frozen inliquid nitrogen and fixed with medium formalin respectively.(3) Two groups weredetected with liver function (AST and T-BIL) and liver pathology. In order to analysis therejection of liver transplantation. serum IL-17, IL-23, TGF-β were detected by ELISA,IL-17expression of liver using immunohistochemical detection and liver IL-17mRNAexpression was detected by real-time RT-qPCR detection. Analyze the expression of IL-17in this model. Results:(1) The success of echinococcus multilocularis infection is85.0%(34/40rats).(2) Survival times after liver transplantation of E.M~+-ROLT group and E.M~--ROLT were23±4.29days and11±1.73days.(3) ALT expression are bothincreased in two groups, but the difference was not statistically significant (P>0.05). bothgroups appear elevated AST and T-BIL, and the difference of both groups werestatistically significant (all P<0.05).(4) At the7th day, E.M~--ROLT group appear strongrejection, and the rejection of E.M~+-ROLT group is not very intensive. And the differenceof Banff97RAI score was statistically significant (P<0.05). The scores were7.67±1.03and5.50±1.28respectively.(5)7days after liver transplantation, the expression of IL-17in E.M~+-ROLT and E.M~--ROLT were detected by different ways:1) immunohistochemistryscore:2.33±1.37and5.00±2.10, the difference was statistically significant, P<0.05.2)ELISA detection value:(16.52±6.41) pg/mL and (28.07±8.43) pg/mL. the differencewas statistically significant, P<0.05.3) real-time RT-PCR detection (IL-17/β-actin relativeexpression levels):0.65±0.27and0.99±0.41, the difference was significant difference,P<0.05.(6) The serum expression of IL-23levels of E.M~+-ROLT group is (422.48±54.47)pg/mL, which is significantly lower than the EM~--ROLT group which expression is(685.18±31.42) pg/mL, and P<0.05.(7) The serum expression of TGF-β levels ofE.M~+-ROLT group is (8.42±2.04) ng/mL, which is higher than the EM~--ROLT group,which expression is (7.67±1.44) ng/mL, but the difference was not statisticallysignificant (P>0.05). Conclusion:(1) Successfully established the subcutaneoussecondary Echinococcus multilocularis infection model of BN rat, which was the basis forsubsequent experimental studies.(2) Successfully established Echinococcus multilocularisinfection in rat liver transplantation model for the study correlation between the parasiteand liver transplantation immunity.(3) Infection with Echinococcus multilocularis canextend the survival of transplanted liver.(4) The low expression of IL-17in serum andtransplanted liver can inhibit the acute transplant rejection of E.m infected rats which wereunderwent liver transplantation, and prolong the survival of transplanted liver andrecipients. |
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