The Experimental Research On The Function Of MiRNA In The Immune Process Of Xenotransplantation In Vitro

Objective:To study the expression of miRNA and the target proteins of Gala(1,3)Gal and NF-κB during the immune process of xenotransplantation in vitro, and control the immune reaction of xenotransplantation effectively.Methods:①Cells of EOMA were cultivated, and then mixed with heterologous serums to build model experiments of xenotransplantation in vitro. It was divided as rat serum group, guinea pig serum group,human serum group and PBS group.②Cells were harvested at the suitable time, and the expressions of miR-146a and miR-155were observed by real-time fluorescence quantitative PCR. The samples were detected using the technology of TaqMan miRNA Assays.③The expression of α(1,3)GT mRNA was detected by RT-PCR. The expression of Gala(1,3)Gal was detected using flow cytometry and the expression of NF-kB (p65) was observed by Western blot.Results:①Compared with PBS group, the expressions of miR-146a and miR-155in rat serum group, guinea pig serum group, and human serum group showed an significant increase(P<0.05). The expression of miR-146a in guinea pig serum group and human serum group was lower than rat serum group. The expressions of miR-146a and miR-155between guinea pig serum group and human serum group had no difference (P>0.05).②Compared with PBS group,The expression of al,3GT mRNA showed an significant increase in guinea pig serum group and human serum group (p<0.05); The expression of α1,3GT mRNA between guinea pig serum group and human serum group had no difference (P>0.05). The expression of al,3GT mRNA between rat serum group and PBS group had no difference (p>0.05)③The expression of Gala(1,3)Gal in the four groups had no difference(.P>0.05).④Compared with PBS group, the expressions of NF-κB (p65) in rat serum group, guinea pig serum group and human serum group showed an significant increase (P<0.05). But there was no difference among the three groups.Conclusions:During the early immune process of xenotransplantation, the expressions of miR-146a and miR-155are upregulated, which play important roles in the immune response. In the immune process of the mouse to rat concordant xenotransplantation, the higher expression of miR-146a play an important role in AVR, which can cause the expressions of NF-kB (p65).But in the immune process of the mouse to guinea pig and the mouse to human disconcordant xenotransplantation, the higher expression of miR-155play an important role in HAR,which can cause the expression of al,3GT mRNA.