【摘要】 目的观察索曲妥林(sotrastaurin或AEB071)对人外周血中单个核细胞(PBMCs)、纯化CD4~+T细胞和CD8~+T细胞分泌细胞因子的抑制作用及其机制。方法采集健康志愿者的静脉血,肝素抗凝,密度梯度离心法分离PBMCs后纯化CD4~+T和CD8~+T细胞,应用抗CD3抗体联合CD28抗体进行刺激,加或不加入AEB071培养后,检测细胞因子的产生、转录因子的表达和细胞的活化与增殖。结果索曲妥林处理不促进外周血T细胞的凋亡,而能抑制T细胞分泌细胞因子IFN-γ、TNF-α和IL-2,且呈时间和剂量依赖性;进一步研究发现,索曲妥林通过下调STAT-1和STAT-4的磷酸化以及T-bet的表达抑制细胞因子的产生。同时经研究发现,索曲妥林处理后抑制T细胞的活化和增殖。结论索曲妥林抑制T细胞转录因子的表达与磷酸化而抑制细胞因子的产生,从而抑制人体免疫功能。为此,进一步研究索曲妥林免疫抑制的机制,对索曲妥林在器官移植、银屑病和溃疡性结肠炎的治疗方面的应用,具有参考价值。更多还原

【Abstract】 To investigate the inhibitory effect and mechanism of sotrastaurin on the production of cytokines,human PBMCs and purified CD4~+ or CD8~+T cells were stimulated with anti-CD3 plus anti-CD28 or with immobilized anti-CD3 and soluble anti-CD28 in the presence or absence of sotrastaurin. The levels of IFN-γ, TNF-α and IL-2 in the culture supernatants were detected by ELISA; the production of cytokines and transcription factors including T-bet, p-STAT-1 as well as p-STAT-4 were analyzed by FACS. Data showed that sotrastaurin lacked sufficient cytotoxicity toward T cells from healthy volunteers but exhibited significant antiproliferation. In addition, sotrastaurin had significant suppressive effects on the production of IFN-γ, TNF-α and IL-2 in a dosetime-dependent manner. Further study revealed that sotrastaurin down-regulated the phosphorylation of STAT4 and STAT1 and the expression of T-bet. All these results indicated that the main mechanism of sotrastaurin inhibiting the cytokine production is down-regulating the phosphorylation of STAT1 and STAT4 and the expression of T-bet.The further study on the mechanism of sotrastaurin immunosuppression is of great significance for its application in the treatment of organ transplantation, psoriasis and ulcerative colitis.更多还原