雷公藤甲素对小鼠淋巴细胞体外活化的抑制作用

The Effects of Triptolide Inhibition of Mouse Lymphocytes Activated in Vitro

目的研究雷公藤甲素(TRD)对小鼠T淋巴细胞活化标志CD69、CD25及细胞因子IL2及IFNγmRNA表达的影响,为阐明其免疫抑制作用提供分子药理学依据。方法无菌分离小鼠淋巴结细胞,加入不同浓度的雷公藤甲素预先孵育1h,用多克隆刺激剂刀豆蛋白(ConA)刺激T细胞活化,利用荧光标记的单克隆体双染技术,流式细胞仪检测TRD对小鼠T细胞CD69、CD25表达的影响;采用半定量RTPCR技术从mRNA水平检测TRD对细胞因子IL2、IFNγmRNA的表达的影响。结果TRD能抑制T细胞早期活化标志CD69和CD25,同时TRD也能抑制IL2及IFNγmRNA的表达。结论TRD对T细胞早期活化分子及细胞因子表达的抑制作用可能是其发挥免疫抑制作用的机制之一。

Objective: To study the effects of triptolidel on murine T lymphocytes activated. Methods: We isolated and cultured the mouse lymphocytes from lymph nodes of mice in vitro. The lymphocytes were pre-treated triptolide for 1 h prior to activation with ConA. After 6 hours stimulation, determined IL-2 and IFN mRNA expression by RT-PCR. CD69 expression of mouse CD3+T cell was determined by multi-color flow cytometric analysis after 6 hours stimulation, and CD25 expression of mouse CD3+T cell was determined by multi-color flow cytometric analysis after 24 hours stimulation. Results: TRD could inhibit IL-2 and IFN-γ mRNA expression. It also inhibited CD69 and CD25 expression of mouse CD3+T cell. Conclusion: the immunosuppressive mechanism of TRD may be related to the inhibitory effects on IL-2 and IFN-γ mRNA expression and the inhibition of CD69 and CD25 expression.