【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RPMI-1640培养液；(2)落新妇甙组,在RPMI-1640培养液中加入落新妇甙15μg/ml；(3)落新妇甙+CTLA-4单克隆抗体组,在RPMI-1640培养液中加入落新妇甙15μg/ml和CTLA-4单克隆抗体9Hi0 30μg/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；逆转录一聚合酶链(RT-PCR)和免疫印记(Western blot)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。更多还原

【Abstract】 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 Cardiomyocytes of BALB/ c mouse and spleen cells of C57BL/6 mouse were separated.The cardiomyocytes(2×10~5/ml)as irri- tation 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/μg/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 sig- nificantly 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.更多还原