Forskolin对小鼠体外T淋巴细胞活化、增殖和周期的影响

Effects of Forskolin on Proliferation Cell-cycle Distribution and Activation of the Murine T Lymphocytes in vitro

目的:研究Forskolin对刀豆蛋白A(Con A)刺激的小鼠CD3+T淋巴细胞活化、增殖和周期的影响,初步探讨其免疫调节作用机制。方法:无菌分离小鼠淋巴结并制备单个淋巴细胞悬液,与不同浓度的Forskolin孵育,运用流式细胞术结合双色荧光抗体染色技术来检测CD3+T细胞早期活化标志CD69的表达水平;用羧基荧光素乙酰乙酸(CFDA-SE)染色技术结合流式细胞术和CELLQuest、Mod FitLT软件,分析T细胞增殖情况;用碘化丙锭(PI)染色技术分析细胞周期分布。结果:加入终浓度为10-7、10-6、10-5M的Forskolin,孵育8 h后,CD3+T细胞CD69的表达水平和平均荧光强度(MFI)与Con A组相比都有显著的降低,且呈梯度依赖性;对Con A刺激的小鼠CD3+T细胞的增殖指数(PI)也有梯度性抑制;对细胞周期的影响表现为:随着Forskolin浓度升高,处于G0/G1期的细胞增加,S期细胞减少,G2/M期的细胞也有少量减少。结论:Forskolin能有效抑制小鼠T淋巴细胞的体外活化和增殖,阻滞细胞从G0/G1期进入S、G2/M期。由此推断Forskolin在某种程度上是一种潜在的免疫抑制剂。

Objective：To investigate the effects of Forskolin on activation,proliferation,and cell-cycle distribution of murine CD3^＋T lymphocytes,and study the mechanisms of its immunosuppressive effect.Methods：Singel cell suspensions were prepared from murine lymph nodes.Fluorescence conjugated monoclonal antibodies and flow cytometry were used to detect the expression of CD69 activated by Con A,the proliferation index of activated mouse T lymphocytes was analyzed by CFDA-SE staining,the distribution of the cell cycle was analyzed by PI staining.Results：Forskolin（10^-7,10^-6,10^-5M）could inhibit both the expression of CD69 on CD3^＋T lymphocytes and T lymphocyte proliferation index stimulated by Con A in a dosedependent manner.The C0/G1 of T lymphocytes increased but the S,G2/M phase decreased.Conclusion：Forskolin can inhibit the activation and proliferation of murine T lymphocytes in vitro,and arreste activated T lymphocytes from G0/G1 to S or G2/M.Forskolin is a potential immunosupressive agent.