目的:内质网应激(ERS)是真核细胞中普遍存在的适应性应激反应,本研究应用ERS特异性诱导剂衣霉素(TM)体外刺激小鼠脾脏树突状细胞(DC),观察DC功能状态的改变情况。方法:分离正常BALB/c小鼠脾脏DC进行体外培养,给予ERS特异性诱导剂TM刺激,观察不同剂量、不同作用时间TM刺激与DC表面共刺激分子表达及分泌功能间的效应关系。结果:TM刺激后,DC表面共刺激分子主要组织相容性复合体(MHC)Ⅱ表达有明显上调(P〈0.05),但未引起CD80、CD86表达水平的明显改变;TM刺激后DC分泌功能有所增强。结论:TM激发的药物性ERS不能有效诱导DC成熟分化。
Objective:To investigate the effect of pharmacological endoplasmic reticulum stress (ERS) induced by tunicamycin (TM) on maturation of splenic dendritic cells (DCs). Methods:DCs were isolated from the spleens of male BALB/c mice, and the cells were stimulated with TM (1 μg/ml for 24 hours, 48 hours or 72 hours; or 0. 1 μg/ml for 48 hours; or 10 μg/ml for 48 hours). After being stimulated, expressions of ERS related molecules were analyzed by SYBR green real-time RT-PCR. Expressions of costimulatory molecules on surface of DC were determined by flow cytometry. Cytokines in cell culture medium were determined by enzyme-linked immunoadsor- bent assay (ELISA). Results:TM induced significant ERS response in mice splenic DC. TM stimulation resulted in elevated cytokine release, and up-regulation of major histocompatibility complex (MHC) II on the surface of DC. However, it failed to induce up-regulation of costimulatory molecules including CD80 and CD86. Conclusions: ERS induced by TM could not effectively induce maturation of DC in vitro.