摘要
目的探讨落新妇甙对混合细胞培养中活化的T细胞产生细胞毒T淋巴细胞相关抗原4（CTLA-4）表达的影响。方法分离BALB／c小鼠心肌细胞（2×10^5／ml）和C57BL／6小鼠脾细胞（1×10^6／ml），前者作为刺激细胞，后者作为反应细胞，进行混合细胞培养。实验分为3组：（1）对照组，混合细胞培养采用RPMI1640培养液；（2）落新妇甙组，在RPMI1640培养液中加入落新妇甙15ttg／ml；（3）落新妇甙＋CTLA-4单克隆抗体组，在RPMI1640培养液中加入落新妇甙15μg／ml和CTLA-4单克隆抗体9H1030μg／ml。原位末端标记（TUNEL）法检测混合细胞培养中T淋巴细胞凋亡情况；逆转录-聚合酶链（RT-PCR）和免疫印记（Westernblot）法检测T细胞CTLA-4表达情况。结果落新妇甙组的T细胞凋亡指数明显高于对照组（73．4％±12．5％ vs 35．1％±9．2％，P〈0．01），T细胞CTLA-4的表达亦显著高于对照组（P〈0．01）。落新妇甙加CTLA-4单克隆抗体组的T细胞凋亡指数和CTLA-4表达与对照组的差异无统计学意义（P〉0．05）。结论落新妇甙诱导心肌细胞排斥反应中活化的T细胞凋亡可能与其增强T细胞中CTLA-4的表达有关。
Objective To investigate the effect of astilbin on expression of CTLA-4 in activated T cells of mouse heart transplantation model with acute rejection. Methods Cardiomyoeytes of BALB/c mouse and spleen cells of C57BL/6 mouse were separated. The eardiomyoeytes （2 × 10^5/ml） as irritation cells and spleen cells （1 × 10^6/ml） as responsers were mixed and cultured. The model of mouse heart transplantation with acute rejection in vitro was established. Three groups were set up： control group, Astilbin （15/xg/ml） group, Astilbin ＋ anti-CTLA-4 mAb 9H10 （30μg/ml） group. Apoptosis of T cells was observed by TUNEL. The expression of CTLA-4 in activated T cells was detected by RT-PCR and Western blot. Results Apoptosis indexes of activated T cells in Astilbin group were significantly higher than those in the control group （73.4 % ± 12.5 % vs 35. 1% ±9. 2 %, P〈0. 01）. The expression of CTLA-4 in Astilbin group was significantly higher than control group （P〈0. 01 ）, but there was no apparently difference between control group and Astilbin ＋ CTLA-4 mAb group （P〈 0. 05）. Conclusion Astilbin induces apoptosis of activated T cells in heart transplantation, which may be partially related to its enhanced expression of CTLA-4.
出处
《中华器官移植杂志》
CAS
CSCD
北大核心
2006年第10期580-582,共3页
Chinese Journal of Organ Transplantation
基金
国家自然科学基金(30300323
30500656)
关键词
心脏移植
落新妇甙
移植物排斥
Heart transplantation
Astilbin
Graft rejection
