淫羊藿提取物对小鼠T淋巴细胞体外活化和增殖的影响  

Effect of extract from Herba Emidemii on activation and proliferation of mouse T lymphocytes in vitro

作　　者：滕菲[1] 曾耀英[1] 黄秀艳[1] 杨志[1] 姚满林[1] 宋兵[1] 李林[1] 

机构地区：[1]暨南大学组织移植与免疫中心,广东广州510632

出　　处：《中草药》2008年第9期1355-1359,共5页Chinese Traditional and Herbal Drugs

基　　金："973"国家重点基础研究项目(2006CB504201和2004CB720100);广东省基金资助项目(2006B36030016);广州市科技局科技攻关重点资助项目(2006Z-E0091)

摘　　要：目的研究淫羊藿水溶性提取物对刀豆蛋白A(ConA)刺激的小鼠T淋巴细胞体外早期活化和增殖的影响。方法无菌分离小鼠各部位淋巴结,制备单细胞悬液,加入不同终质量浓度(5、25、50mg/L)的淫羊藿提取物,以MTT法检测药物对T细胞活性的影响;利用流式细胞术(FCM)结合双色荧光抗体染色技术检测早期活化标志CD69分子的表达;运用流式细胞术结合活体染料CFDA-SE染色技术和荧光抗体染色技术检测T细胞增殖。结果MTT法检测淫羊藿提取物在50mg/L时细胞存活率达52.36%。ConA作用6h后,对照组CD69+表达率为(4.61±0.85)%,ConA组CD69+T细胞的比率上升至(38.30±2.54)%,淫羊藿提取物在终质量浓度5、25、50mg/L时均抑制ConA诱导的CD3+T淋巴细胞CD69的表达(P<0.01),其表达率分别下调为(36.16±2.14)%,(30.5±1.72)%,(15.99±0.87)%。培养48、72后,对照组增殖指数(PI)分别为1.01±0.01和1.05±0.01,ConA组PI分别达到1.42±0.01、2.32±0.01,淫羊藿提取物在终质量浓度5、25、50mg/L均抑制T淋巴细胞增殖(P<0.01),48h时PI分别为1.28±0.01、1.26±0.01、1.21±0.01;72h时PI分别为2.30±0.03、2.13±0.02、2.10±0.01。结论淫羊藿提取物能明显抑制ConA刺激的小鼠T淋巴细胞的早期活化和增殖,并呈剂量依赖性。Objective To study the effect of water extract from Herba Ernidemii on the early activation and proliferation of mouse T lymphocytes stimulated by concanvalin A （Con A） in vitro. Methods Single cell suspension was prepared from mouse lymphoid nodes in germ free condition. The drug toxicity of water extract from Herba Emidemii （5, 25, and 50 mg/L） on T lymphocytes was measured by MTT. The expression of CD69, the marker of early activation of T lymphoeytes was measured by flow eytometry （FCM） combined with two-colour immunofluorescent staining of cell surface antigen. The proliferation of T lymphocytes was measured by FCM combined with carboxyl fluorescin diacetate succinmidyl ester （CFDA-SE） staining and immunofluorescent staining. Results The survival ratio of T lymphocytes in 50 mg/L was 52.36%. After 6 h culture, the expression rate of CD69^＋T cell in control group was （4.610±.85）%, that in Con A group was （38.30±2.54）%, water extrat from Herba Emidmii （5, 25, and 50 mg/L） inhibited the expression of CD69 in CD3^＋T lymphocytes stimulated by Con A （P〈0.01）, the expression rates were （36. 16±2. 14）%, （30. 5±1.72）%, and （15.99±0.87）%, respectively. After 48 and 72 h culture, the proliferation index （PI） of control group were 1.01 ±0. 01 and 1.05±0. 01, of Con A group were 1.42±0. 01 and 2.32±0.01, respectively. Water extract from Herba Emidmii （5,25, and 50 mg/L） inhibited the proliferation of T lymphocytes （P〈0. 01）, the PI of 48 h were 1.28±0.01, 1.26±0.01, 1.21±0.01; the PI of 72 h were 2.30±0.03, 2.13±0.02, 2.10±0.01, respectively. Conclusion Water extract from Herba Ernidrnii could significantly inhibit Con A-induced early activation and proliferation of mouse T lymphocytes in vitro in a dose dependent manner.

关 键 词：淫羊藿提取物 T淋巴细胞 活化 增殖 

分 类 号：R286.95[医药卫生—中药学]