染料木黄酮与环孢菌素A联合用药对小鼠淋巴细胞体外免疫应答的影响  

Effects of combination of Genistein with Cyclosporin A on immune response of mouse lymphocytes in vitro

作　　者：叶雪仪[1] 曾耀英[1] 肇静娴[1] 狄静芳[1] 贺芳[1] 周建国[1] 赵令斋[1] 

机构地区：[1]暨南大学组织移植与免疫实验中心,广东广州510632

出　　处：《暨南大学学报（自然科学与医学版）》2007年第3期322-326,共5页Journal of Jinan University(Natural Science & Medicine Edition)

基　　金：国家自然科学重点基金资助项目(30230350);广东省自然科学基金资助项目(5300413)

摘　　要：研究染料木黄酮(Gen)与环孢菌素A(CsA)联合用药对小鼠体外淋巴细胞增殖、周期及混合淋巴细胞反应的影响,从而为将Gen开发为免疫干预药物以降低CsA的临床用量提供理论依据.以刀豆蛋白A(ConA)诱导小鼠淋巴细胞增殖,通过CFDA-SE染色流式细胞术检测荧光强度的变化,用相关软件分析其增殖指数;以碘化丙锭(PI)染色分析细胞周期的分布情况;MTT法测定体外混合淋巴细胞反应(MLR).结果显示,细胞培养48 h后,ConA诱导下小鼠淋巴细胞与空白对照组相比有明显增殖(P<0.01),5、10μmol/L Gen以及0.02μmol/L CsA均可抑制这种增殖作用,二者联合用药后,这种抑制增殖的作用加强(P<0.01).对细胞周期分析显示,Gen和CsA抑制细胞由G0/G1期向S期转化.MTT法测定的结果显示,二者联合用药后Q值分别为1.82、1.39,对MLR呈协同抑制效应.以上结果表明与单剂量Gen、CsA相比,二者联合使用对ConA诱导的淋巴细胞增殖、细胞周期的阻滞、体外混合淋巴细胞反应的抑制作用更加明显,呈协同效应.To investigate effects of the combination of Genistein （Gen） with Cyclosporin A （CsA） on the mouse lymphocytes proliferation and cell-cycle, as well as double-way mixed lymphocyte reaction （MLR）, in order to develop Gen as an immuno-intervention reagent that reduce the clinical CsA. Stimulated with Concanavalin A （Con A） dosage of , the change of fluorescence intensity was determined by carboxyl fluorescin diacetate succinmidyl ester （CFDA-SE） flowcytometry, and related software was applied to analyze the proliferation index. The cell-cycle distribution was analyzed by propidium iodide （PI） staining. And MLR was detected by MTT colorimetry. It was found that after 48 h culture, the ConA group exerted visible proliferation effect compared with the control group （P 〈 0. 01 ）. The proliferation was suppressed by 5, 10 μmol/L Gen and 0. 02 μmol/L CsA alone. When combining Gen with CsA at the upper concentrations, this inhibitory effect was enhanced （P 〈 0. 01 ）. The cell-cycle ananlysis showed that they inhibited the transition of cells from G0/G1 phase to S phase. MtT colorimetry indicated that the combination showed synergy effect in MLR, with Q value equaled to 1.82, 1.39, respectively. These results suggest that combining Gen with CsA performs more evident antiproliferative response, blockage in cell-cycle distribution and inhibition of MLR. Moreover, these effects are synergistical.

关 键 词：染料木黄酮 环孢菌素A 增殖 周期 混合淋巴细胞反应 

分 类 号：R363[医药卫生—病理学]