| Summary: | 碩士 === 輔仁大學 === 生命科學系碩士班 === 100 === The allogeneic bone marrow transplantation (BMTx) is the essential therapy used to treat non-hematologic and hematologic diseases. There is nearly 30-50% occurrence of acute graft-versus-host disease (aGvHD) after allogeneic BMTx. GvHD is a T cell-mediated acute inflammation caused by donor T cell attacking host tissue. Thus, one of the practicable strategies to alleviate GvHD after allogeneic BMTx is to develop immunosuppressive drugs to T cell activation with low toxicity. Piperlactam S, is a natural compound extracted from Piper kadsura. Previous research have showed that Piperlactam S was proved the immunosuppressive bioactivities on the proliferation and cytokines expression of human primary T cells, human neutrophils, and mice macrophage Raw 264.7. In previous study, Piperlactam S showed immunosuppressive bioactivities on mouse spleen T cell proliferation after anti-CD3/CD28 mAb stimulation and the production of the T helper-type cytokines like IFN-γ, IL-2, and IL-10, and especially mRNA expression of NF-AT, the specific transcription factor of T cell activation. But until now, it is not clear about the immunosuppressive mechanisms of Piperlactam S to apply on GvHD prevention. Based on these, this study goals of this are to analyze the Piperlactam S in vitro and setting up the mice GvHD animal model to evaluate the possibility of the application between GvHD and Piperlactam S. The in vitro results indicated (1) Piperlactam S possessed the immunosuppressive bioactivities on the proliferation of murine spleen-T cells by anti CD3/CD28 mAb activation, which I confirm again. The IC50 of Piperlactam S is 8.72μM. But at 40μM, there was about 30% cytotoxicity. (2) In the Jurkat T reporter gene system which transfected NF-κB and NF-AT genes treated with Piperlatam S, there was immunosuppressive bioactivity on the expression of NF-κB and NF-AT, in dose dependent manner. (3) Piperlactam S possessed immunosuppressive bioactivities on the proliferation of T cells activated by alloantigen, in the dose dependent manner. The IC50 is 11.88μM, but there was cytotoxicity more then 40μM, (4) Piperlactam S presented dose-dependent inhibition activities to mRNA and proteins of IL-2, IFN-γ, and IL-10. (5)Piperlactam S inhibited the inflammatory cytokine level of IL-17. (6) There was no affection on the mRNA expression of AP-1 and NF-AT of activated T cells by alloantigen when Piperlactam S co-culture with. (7) Piperlactam S inhibited the dephosphorylation of phosphor-NF-AT but there was no affection on the protein level of NF-AT. (8) In murine acute GvHD model, the proper dosage setting after the programs to cytotoxicity and tolerance of Piperlactam S could promoted the survive of the mice after allogeneic BMTx with GvHD syndromes. Based on these results, Piperlactam S extracted from Piper kadsura, was provide with the immunosuppressive bioactivities for alloantigen-activated T cell and reduced the reaction of GvHD.
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