伪石蒜碱抑制活化T细胞增殖与功能的机制研究  

Study on the Mechanism of Pseudolycorine Inhibiting the Proliferation and Function of Activated T Cell

作　　者：吴利红 邓弘仙 张云惠 黄玲 范诗逸 蒋海静 赖翼[3] 罗兴燕[4] 刘阳[4] 杨淑霞[4] WU Lihong;DENG Hongxian;ZHANG Yunhui;HUANG Ling;FAN Shiyi;JIANG Haijing;LAI Yi;LUO Xingyan;LIU Yang;YANG Shuxi(Chengdu Medical College,School of Pharmacy,Chengdu 610500,China;Chengdu Medical College,School of Clinical Medicine,Chengdu 610500,China;Chengdu Medical College,School of Laboratory Medicine,Chengdu 610500,China;Chengdu Medical College,Research Center,Chengdu 610500,China)

机构地区：[1]成都医学院药学院,成都610500 [2]成都医学院临床医学院,成都610500 [3]成都医学院检验医学院,成都610500 [4]成都医学院科研中心,成都610500

出　　处：《中国现代应用药学》2020年第15期1818-1824,共7页Chinese Journal of Modern Applied Pharmacy

基　　金：四川省教育厅科研计划重点项目(18ZA0143);植物化学与西部植物资源持续利用国家重点实验室开放项目(P2018-KF03);成都医学院国家级、省级大学生创新创业训练计划项目(201913705049)。

摘　　要：目的探究伪石蒜碱抑制活化T细胞增殖与功能的机制。方法密度梯度离心法和免疫磁珠法分离纯化T细胞,抗CD3/CD28或植物凝集素(phytohemagglutinin,PHA)活化T细胞。流式细胞术检测细胞增殖、细胞凋亡、CD25表达及细胞周期;ELISA检测细胞因子IL-2、IL-6、IL-17A、IFN-γ的分泌水平。结果伪石蒜碱抑制抗CD3/CD28或PHA活化T细胞增殖,IC50分别为(0.97±0.22)μmol·L-1和(0.82±0.07)μmol·L-1。在完全抑制活化T细胞增殖的浓度下,伪石蒜碱不诱导活化T细胞凋亡,且不对静息T细胞的细胞活力产生显著影响。伪石蒜碱不影响活化T细胞表达CD25和分泌IL-2,但阻滞细胞周期于G0/G1期。伪石蒜碱显著抑制IL-6、IL-17A、IFN-γ的分泌。结论伪石蒜碱不影响T细胞的活化,但通过阻滞细胞周期于G0/G1期抑制活化T细胞的增殖,提示伪石蒜碱有望成为先导化合物用于开发新型免疫抑制剂。OBJECTIVE To explore the inhibitory effects of pseudolycorine on activated T cell proliferation and its mechanism.METHODS Human T cells were isolated and purified by density gradient centrifugation and the immunomagnetic microbeads,and activated by anti-CD3/CD28 mAbs or phytohemagglutinin(PHA).Cell proliferation,apoptosis,CD25 expression and cell cycle were detected by flow cytometry.Level of cytokines IL-2,IL-6,IL-17A and IFN-γwere measured by ELISA.RESULTS Pseudolycorine inhibited human T cell proliferation with anti-CD3/CD28 mAbs stimulation with an IC50 value of(0.97±0.22)μmol·L-1 and PHA stimulation with an IC50 value of(0.82±0.07)μmol·L-1.Pseudolycorine did not induce apoptosis of activated T cells and had no significant effect on the cell viability of resting T cells with the same concentration of inhibiting activated T cell proliferation.Pseudolycorine did not affect CD25 and IL-2 expression,but induced cell cycle arrest in G0/G1 phase.Pseudolycorine significantly inhibited IL-6,IL-17 and IFN-γexpression.CONCLUSION Pseudolycorine does not affect the activation of T cells,but inhibits the proliferation of T cells by blocking the cell cycle in G0/G1 phase.Pseudolycorine represents a potential lead compound for the design and development of new immunosuppressive drugs.

关 键 词：免疫抑制剂 T细胞增殖 伪石蒜碱 细胞因子 

分 类 号：R285.5[医药卫生—中药学]