白芍总苷抑制小鼠T淋巴细胞体外增殖促进活化诱导细胞死亡  

Total glucosides of paeony inhibits proliferation and promotes activation-induced cell death of mouse T cells in vitro

作　　者：熊传锋 齐杰莹 邓蓉 谢丽芬[1] 李长征[1] 聂晓莉[1] XIONG Chuanfeng;QI Jieying;DENG Rong;XIE Lifen;LI Changzhang;NIE Xiaoli(School of Traditional Chinese Medicine,Southern Medical University,Guangzhou 510515,China;Department of Traditional Chinese Medicine,Nanfang Hospital,Southern Medical University,Guangzhou 510515,China)

机构地区：[1]南方医科大学1中医药学院,2南方医院中医科,广东广州510515

出　　处：《南方医科大学学报》2020年第1期118-124,共7页Journal of Southern Medical University

基　　金：国家自然科学基金(81774038);广东省自然科学基金(2015A030313300);广东省科技计划项目(2014A020221073);广东省中医药局项目(20181168);广州市科技计划项目(201607010224)~~

摘　　要：目的研究白芍总苷(TGP)对小鼠T淋巴细胞体外增殖及活化诱导细胞死亡(AICD)的影响,并初探其免疫抑制机制。方法利用免疫磁珠分选技术,无菌分离小鼠脾脏T淋巴细胞,按照TGP溶液浓度(0、50、100、200μg/mL)设为空白组、TGP低中高剂量组,anti-CD3/CD28刺激T淋巴细胞活化及增殖。流式细胞仪检测细胞早期活化标志、细胞活力及CFSE标记的T淋巴细胞增殖;RT-PCR检测Fas/FasL m RNA水平;流式细胞仪检测活化T淋巴细胞表面Fas/FasL蛋白表达量;Western blot检测细胞凋亡相关蛋白Bcl-2的表达。结果 (1)与空白组相比,TGP中高剂量组在用药48 h后活细胞数目显著减少,活细胞比例明显下降(P<0.001);(2)TGP组给药处理48 h后,T淋巴细胞分裂代数及已分裂比例显著降低,且抑制作用具有药物浓度依赖性(P<0.001);(3)TGP处理组T淋巴细胞24 h后Fas mRNA表达量显著上调,并伴随着T淋巴细胞表面Fas蛋白的表达上调及胞内Bcl-2表达下调(P<0.01)。结论 TGP显著抑制T淋巴细胞增殖,并可通过上调Fas、下调Bcl-2的表达促进活化诱导的T淋巴细胞死亡(AICD)。Objective To investigate the effects of total glucosides of paeony(TGP) on the proliferation and activation-induced cell death of mouse T cells and the mechanism underlying the immunosuppressive effects of TGP. Methods Purified total T cells isolated from the spleen of C57 BL/6 mice were treated with TGP at 0, 50, 100, or 200 μg/mL and stimulated by anti-CD3/CD28. Flow cytometry was performed to detect the cell death and the proliferation of CFSE-labeled T cells. The expression of Fas/FasL mRNA was detected using RT-PCR, and flow cytometry was used to analyze the expression of Fas/FasL proteins on activated T cells. Western blotting was used to detect the expression of Bcl-2 in the cells. Results TGP treatment for 48 h significantly reduced the total number and percentage of viable T cells and dose-dependently lowered the percentage of divided T cells. TGP treatment obviously up-regulated the cellular expression of Fas mRNA, enhanced Fas expression on the surface of the T cells, and decreased the expression level of Bcl-2 protein in the cells. Conclusion TGP significantly inhibits proliferation and promotes activation-induced cell death of mouse T cell by increasing the expression of Fas and downregulating the expression of Bcl-2.

关 键 词：白芍总苷 细胞增殖 活化诱导细胞死亡 FAS/FASL BCL-2 

分 类 号：R285.5[医药卫生—中药学]