咖啡酸苯乙酯对小鼠CD3^+T细胞体外活化和增殖的影响  

Effects of caffeic acid phenethyl ester on activation and proliferation of murine CD3^+ T lymphocytes in vitro

作　　者：唐先高[1] 曾耀英[1] 黄秀艳[1] 周建国[1] 

机构地区：[1]暨南大学组织移植与免疫实验中心,广东广州510632

出　　处：《中国药理学通报》2008年第4期472-477,共6页Chinese Pharmacological Bulletin

基　　金：国家重点基础研究发展计划(973计划)资助项目(No2006CB504200;2004CB720100);广东省基金资助项目(No2006B36030016);广州市科技局科技攻关重点资助项目(No2006Z-E0091)

摘　　要：目的研究咖啡酸苯乙酯(caffeic acid phenethyl ester,CAPE)对小鼠CD3+T淋巴细胞活化及增殖的影响,探讨其作用机制。方法无菌分离小鼠淋巴结淋巴细胞,制备淋巴细胞悬液。在淋巴细胞活化实验中,分别与不同浓度的咖啡酸苯乙酯预先孵育2h后,加入促分裂原伴刀豆蛋白A(ConA),运用荧光抗体染色技术结合流式细胞术,8h后,检测T淋巴细胞早期活化标记分子CD69的表达情况;24h后,检测T淋巴细胞中期活化标记分子CD25的表达情况;在淋巴细胞增殖试验中,运用CFDA-SE标记法检测T淋巴细胞刺激48h后的增殖情况。结果CAPE(0·5、1、5、10mg·L-1)能够明显地抑制ConA刺激的小鼠淋巴细胞活化,且呈浓度依赖性;CAPE(0·5、1、5、10mg·L-1)能够明显地抑制ConA刺激的小鼠淋巴细胞增殖,且呈浓度依赖性。结论CAPE对小鼠CD3+T淋巴细胞的体外活化和增殖具有抑制作用,其作用机制可能通过同时抑制PLC-γ信号途径和MAP激酶途径,由此推理CAPE是一种潜在的有效的免疫抑制剂。Aim To investigate the effects of caffeic acid phenethyl ester （CAPE） on activation and proliferation of murine CD3^＋ T lymphocytes, and to study the mechanisms of its immunomodulative effects. Methods Lymphocytes suspension was isolated and prepared sterilely from murine lymph nodes, and was incubated with different concentrations of CAPE for 2 h, then polyclonal activator Con A was added to activate the lymphocytes; double-fluorescence staining and flow cytometry were used to detect CD69 and CD25 expressions to evaluate activation level of CD3^＋ T lymphocyte, and CFDA-SE staining plus flow cytometry were used to analyze CD3^＋ T lymphocyte proliferation.Results CAPE （0.5, 1,5, 10mg· L^-1） could inhibit the expression of CD69 and CD25 of CD3^＋ T lymphocytes in a dose-dependent manner, and it also could inhibit T lymphocyte proliferation stimulated by Con A in a dose-dependent manner. Conclusion In vitro, CAPE could exert notably inhibition on activation and proliferation of murine CD3^＋ T lymphocytes, and its immunomodulative effects might result from affecting PLC-γ signaling and MAPK-signaling pathways. So CAPE was a potentially effective immunoinhibitory agent.

关 键 词：咖啡酸苯乙酯 T淋巴细胞 活化 增殖 流式细胞术 

分 类 号：R-332[医药卫生] R329.24