| Objective: 1.To explore the induction of immune tolerance by the injection of MBP into hepatic portal vein and the role of Kupffer cells in this process.We value this result by detected some inflammation factors.2.SD rat model of SBI was established by injecting auto-brain cell homogenate and MBP into the portal vein to establish the specific immune tolerance.To investigate the effect of liver immune tolerance on brain injury.Methods: 1.32 male SD rats were divided into four groups according to random number table,8 in each group: A1 group: sham group,B1 group: normal saline control group,C1 group: MBP experiment group,D1 group: Gd Cl3 were treatment group(injected Gd Cl3 into rat tail vein 24 h before the portal vein injection).Inject MBP into rat tail vein 10 days after the portal vein injection.MBP antibody 、 TGF-β1 were detected by ELISA,CD4~+/CD8~+ T cells ratio in peripheral blood detected by FCM 24 h and 48 h after the rat tail vein injection.48 h after rat tail vein injection,Fas L were measured by Real-time Polymerase Chain Reaction(RT-PCR).2.32 males SD rats were divided into four groups according to random number table,8 in each group: A2 group: sham group,B2 group: normal saline control group,C2 group: MBP treatment group,D2 group: auto-cerebral cells treatment group.A standardized SBI model was used in B2、C2 and D2 groups,as described in previous reports,A2 group no durotomy.After SBI and sham surgery,all rats developed laparotomy to exposed hepatic portal vein and injected into 0.5ml normal saline、MBP and auto-cerebral cells separately in B2、C2 and D2 groups.Function outcomes was evaluated using MNSS;IL-2,IL-4were detected by ELISA;CD4~+/CD8~+ T ratio in peripheral blood detected by FCM;apoptosis of nerve cells around the surgery wound detected by immunofluorescence at 1,3,7,14,21 d after SBI.Results: 1.Compared with group A1,B1 and D1,the concentration of MBP antibody were extraordinarily lower in group C1(P<0.05),meanwhile CD4~+/CD8~+ T ratio were obviously lower in group C1(P<0.05),at 24 h and 48 h after rat tail vein injection,the gene expression of Fas L were notably higher than that in group C1(P<0.05),at 48 h after rat tail vein injection.Compared with group A1 and B1,the concentration of TGF-β1 were obviously higher than that in group C1 and D1(P<0.05),and there were no difference between group C1 and D1 at 24 h and 48 h after rat tail vein injection.There was no difference about the same indicator,at different time points,between group A1 and B1.2.The modified neurological severity score in C2,D2 group higher than that in A2 group at 1,3,7,14 d,and B2 group higher than A2 group at all the time points,moreover the MNSS in C2 and D2 group lower than that in B2 group(P<0.05)at 7,14,21 d after SBI.D2 and C2 groups show significant lower IL-2 cytokine level and CD4~+/CD8~+ T ratio and higher IL-4 cytokine level compare with A2 and B2 group and at the same time;D2 group better than C2 group at 7,14 d after SBI(P<0.05).At 21 d after SBI,C2,D2 group have higher IL-4 cytokine level than A2,B2 group moreover D2 group better than C2 group(P<0.05),C2 and D2 group have lower nerve cells apoptosis than B2 group(P<0.05).Conclusion:1.Through hepatic portal vein injection of MBP can induce the establishment of specific immune tolerance of MBP,reduce the body’s immune system secondary immune attack against MBP antigen,and MBP accounted for 30% of the brain antigen,therefore has the possibility to protect the damaged brain tissue;liver Kuppffer cells play an important role in liver immune tolerance.2.The establishment of specific immune tolerance by its own brain antigen and MBP of hepatic portal vein injection,nerve inflammation,reduce the apoptosis of nerve cells reduced after SBI,is conducive to the recovery of neurological function,has a good curative effect on surgical brain injury and provide a new way for treatment of brain injury surgery.3.The specific immune tolerance can be established by injecting autologous brain antigen and MBP into the portal vein,and the therapeutic effect of the mixed brain antigen is better than that of the single brain antigen MBP.Considering the two factors,first,the establishment of a variety of antigens induce immune tolerance to self antigen mixed brain,is better than a single antigen immune tolerance;second,“bystander suppression” was activated by tolerogen and resulted in the suppression of immune responses to other antigens Specific reasons and mechanisms still need further study,but also for our follow-up research direction. |
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