雷公藤甲素治疗异基因骨髓移植后aGVHD的实验研究

An experimental study of the inhibitory effect of TWH on acute GVHD after allograft transplantation in mice

目的:探索雷公藤甲素(TWH)对异基因HSCT中aGVHD的治疗作用及其机制,并研究雷公藤甲素对移植物抗肿瘤(GVT)作用的影响和对移植物植入的影响。方法:(1)以B6为供者,致死性照射的CB6F1为受者,建立基因半相合移植动物模型;以CB6F1为受者,皮下接种S180细胞并照射,建立基因半相合移植荷瘤aGVHD模型;BALB/c小鼠背部皮下注射S180细胞并照射,建立同基因移植荷瘤动物模型。各组动物分别给予不同剂量TWH及CsA。观察TWH对GVHD、GVT的影响。(2)MTT法测定TWH抑制脾淋巴细胞增殖作用和TWH的体外抑瘤作用。(3)ELISA法测定移植后动物血清IL-4、IFN-γ水平。(4)甲基纤维素半固体培养基检测CFU-GM生成率。结果:(1)小剂量TWH(<1μg/d)降低aGVHD发病率,延长动物生存时间;小剂量TWH与小剂量CsA在预防GVHD方面有协同作用。(2)TWH在抑制aGVHD的同时,保留GVT作用,延长了荷瘤动物生存时间,而无GVHD的同基因移植荷瘤动物生存期缩短。(3)TWH可抑制脾细胞增殖,降低血清IFN-γ水平。(4)一定剂量TWH(<1μg/d)不影响干细胞植入和CFU-GM的产率。结论:TWH具有抑制aGVHD保留GVT的作用,可与CsA协同作用预防异基因HSCT中aGVHD。

Objective：To investigate the therapeutic mechanism of tripterygium wilfordii hook（TWH） in treating acute graft-versus-host disease（GVHD） after allograft transplantation, and to study the effect TWH on stem-cell engraftment, and on graft antitumor capacity .Methods： （1） In order to eonstmct semi-syngeneic transplantation model,bone marrow ceils and spleen ceils from healthy C57BL/6（136） mice were transplantated into lethally irradiated healthy CB6F1 mice. For the eonstruefion of semi-syngeneie transplantation models for tumor-burden mice,S180 sarcoma cell line cells were subcutaneous injected into CB6F1 mice. After given lethally irradiation, these mice were transplanted bone marrow cells and spleen cells from healthy 136 mice. For syngeneie transplantation models,S180 sarcoma cell line cells were subcutaneous injected into BALBIc mice. After given lethally irradiated, these mice were transplanted BM cells and spleen cells from BALB/c mice. TWH with or without CsA in different dosage were given to each group mice to see their effect on acute GVHD,graft antitumor effect and on graft engraftment. （2）MTT method was used to detect the inhibitory effect of TWH on the proliferation of spleen cells and tumor cells in vitor. （3）ELISA method was used to measure the serum lever of IL-4 and IFN-γ. （4）Methyl cellular semisolid culturing was done to observe the number of CFU-GM. Resuits： （1）Low dose TWH（〈1 μ/d） could reduce the morbidity induced by aGVHD,therefore prolong the survival time of the mice. （2）TWH and CsA in low doses lind synergic effect in preventing aGVHD in there experimental mice. （3）TWH could inhibit aGVHD partially but retain graft-versus-tumor（GVT） effect. The survived time of tumor-burden mice with slight aGVHD after semi-syngeneie transplantation was longer that of tumor burden mice with on aGVHD after syngeneie transplantation. （4）TWH could inhibit the proliferation of spleen cell and the production of IFN-γ. （5）Low dose THW（〈1μg/d） had no effect on stem cell engrafting and on the number of CFU-GM. Conclusion：TWH could prevent aGVHD but retain GVT after allo-HSCT,which could be enhanced by the combined used of TWH with CsA.