Study on the role of KPNB1 inhibitor in mouse transplant rejection
Objective:To investigate the role of inhibitor of nuclear import-43(INI-43)as a karyopherin beta 1(KPNB1)inhibitor in mouse transplant rejection.Methods:Firstly,in vitro experiments were conducted to divide Jurkat T cells into four groups:blank group,activated control group,INI-43 low-dose group,and INI-43 high-dose group;Chromatin immunoprecipitation technique was used to detect the effect of INI-43 on the binding level of nuclear factor of activated T cells(NFAT)and interleukin-2(IL-2)promoter;Dual luciferase assay was used to detect the effect of INI-43 on the transcriptional activity of IL-2 promoter reporter genes.Subsequent-ly,animal experiments were conducted to divide the mice into five groups:sham surgery group,model control group,cyclosporine A(CsA)group,low-dose INI-43 group,and high-dose INI-43 group.Skin transplantation surgery was performed on the experimental mice,and the survival time of the transplant was recorded;Flow cytometry was used to detect the inhibitory effect of INI-43 on the sur-face activation molecules CD25 and CD107a of mouse T cells;Enzyme linked immunosorbent assay was used to measure the levels of IL-2 and interferon gamma(IFN-γ)in peripheral blood of mice.Results:Western blotting showed that compared with the activated control group,the levels of NFAT in the nuclear of both low dose and high dose INI-43 groups decreased(Z=-1.96,t=20.88,both P<0.05),and the binding levels of NFAT and IL-2 promoter in the nuclei decreased(t=5.72,12.51,both P<0.01).Compared with the activated controlgroup,the transcription activity of the wild-type IL-2 promoter reporter gene was reduced in the low-dose and high-dose INI-43 groups(t=12.69,27.30,both P<0.001).The survival time of the transplant was prolonged(Z=-3.71,-3.89,both P<0.001).Compared with the model control group,the expression level of T cell CD25 decreased in the high-dose INI-43 group(t=2.10,P<0.05).The expression levels of CD107a(Z=-3.40,-3.78,both P<0.01),IL-2(t=5.35,9.07,both P<0.01)and IFN-γ(t=6.16,6.81,both P<0.01)in peripheral blood in the low-dose and high-dose INI-43 groups decreased.Conclusion:KPNB1 inhibitor INI-43 can inhibit NFAT transnuclear transport,suppress T cell activation in transplanted mice,and alleviate rejection reactions.
万方数据
维普
