淫羊藿素、脱水淫羊藿素对T淋巴细胞的比较研究  

Comparison of effects of icaritin and anhydroicaritin on lymphocytes

作　　者：赖新强[1] 黄秀艳[2] 曾耀英[2] 

机构地区：[1]暨南大学分析测试中心,广东广州510632 [2]暨南大学生命科学技术学院组织移植与免疫中心,广东广州510632

出　　处：《暨南大学学报（自然科学与医学版）》2013年第3期353-358,共6页Journal of Jinan University(Natural Science & Medicine Edition)

基　　金：国家自然科学基金项目(31200667)

摘　　要：通过分离制备小鼠淋巴细胞;CCK-8法检测脱水淫羊藿素(AHI)、淫羊藿素(ICA)对淋巴细胞的毒性;流式细胞术(FCM)结合双色免疫荧光染色技术检测AHI、ICA对con A刺激T淋巴细胞早期活化标志CD69,中期活化标志CD25的表达情况;利用活性染料CFSE,结合流式细胞术分析AHI、ICA对T淋巴细胞增殖的影响;流式细胞术分析AHI、ICA对T淋巴细胞周期的影响;流式细胞术实时分析AHI、ICA对T淋巴细胞钙离子内流的影响;使用流式蛋白定量检测技术(CBA)检测AHI、ICA对T淋巴细胞分泌细胞因子的影响.结果显示AHI与ICA对活化的小鼠T淋巴细胞具有明显的免疫抑制作用,特别是ICA,能明显抑制经con A刺激的T淋巴细胞早中期活化、增殖、钙离子内流,并阻滞细胞周期在G0/G1期,以及分泌IL-6、IL-10、IFN-γ、TNF和IL-12p70细胞因子;ICA比AHI对活化的小鼠T淋巴细胞具有更明显的免疫抑制作用.Mouse lymphocytes were isolated. The drug toxicology of different concentrations of AHI/ ICA on lymphocytes was measured by CCK-8 assay. The expression of CD69 and CD25, which were the markers of activation of T lymphocytes, was measured by FCM combined with two-color immunofluores- cent staining of cell surface antigen. The proliferation of T lymphocytes was measured by FCM combined with live dye CFSE. The cell cycle of T lymphocytes was measured by FCM. The Calcium influx of T lymphocytes was measured by FCM＇ s time function. CBA kit was used to detect the production of cyto- kines of lymphocytes stimulated by con A. AHI and ICA decreased proportions of CD69 and CD25 on con A-stimulated lymphocytes in a dose-dependent manner. Both reduced the proliferation of T lymphocytes, and the influx of Calcium. AHI and ICA caused a GO/G1 cell cycle arrest on con A-activated lympho- cytes. Furthermore, both downregulated the secretion of cytokines （IL-6, IL-10, IFN-γ, TNF, IL- 12p70） of con A-activated lymphocytes. Furthermore, ICA exhibited more immunosuppressive effects than AHI on mouse T lymphocytes.

关 键 词：淫羊藿素 脱水淫羊藿素 T淋巴细胞 免疫抑制 

分 类 号：R285.5[医药卫生—中药学]