杨梅素对淋巴细胞活化及增殖的影响

目的研究杨梅素(myricetin)对小鼠T细胞活化及增殖的影响,探讨其作为免疫调节药物开发的可能性并提供相关实验依据。方法无菌分离小鼠淋巴结细胞,加入不同浓度的杨梅素预先孵育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抗体双染技术,流式细胞仪检测杨梅素对小鼠CD3+T细胞CD69表达的影响;采用3H-TdR参入法检测杨梅素对淋巴细胞增殖的影响,采用半定量RT-PCR技术从mRNA水平检测杨梅素对IL-2mRNA表达的影响。采用ELISPOT检测杨梅素对IFN-γ分泌的影响。结果杨梅素能抑制T细胞早期活化标志CD69的表达,并能抑制淋巴细胞增殖反应,同时对淋巴细胞活化后IL-2 mRNA的表达及IFN-γ的分泌也有抑制作用。结论杨梅素对淋巴细胞的活化增殖反应具有抑制作用。

The effects of myrictin on inhibition of mouse lymphocytes activated in vitro

Aim To study the effects of myricetin on murine lymphocytes activated.Methods We isolated and cultured the mouse lymphocytes from lymph nodes of mice in vitro.The lymphocytes were pre-treated with myricetin for 1 h prior to activation with ConA.After 6 hour stimulation,IL-2 mRNA expression was determined using semi-quantitative RT-PCR.CD69 expression of mouse CD3~+ T cell was determined using multi-color flow cytometric analysis.T cells proliferation was detected using()~3H-TdR incorporation and IFN-γ secretion is determined with ELISPOT assay.Results 5～100 μmol·L~(-1) myricetin inhibited CD69 expression of mouse CD3~+ T cell,12.5～100 μmol·L~(-1) myricetin inhibited proliferation of mouse lymphocytes.100 μmol·L~(-1) myricetin completely inhibited IL-2 mRNA expression of mouse lymphocyte,50 μmol·L~(-1) myricetin and 10 μmol·L~(-1) myricetin also inhibited IL-2 mRNA expression.1 μmol·L~(-1) and 10 μmol·L~(-1) myricetin also inhibited IFN-γ secretion stimulated by ConA.Conclusion Myricetin may has the inhibitory effects on lymphocyte activation and proliferation.