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中文摘要：
目的观察高迁移率族蛋白B1（HMGB1）对大鼠脾脏树突细胞（DC）作用的受体机制。方法分离正常Wistar大鼠脾脏DC,置于96孔培养板（1×105/孔）,HMGB1刺激后采用激光扫描共聚焦显微镜和流式细胞术检测DC表面晚期糖基化终末产物受体（RAGE）的表达强度。同时观察抗RAGE抗体对HMGB1刺激后DC表面共刺激分子CD80、CD86和主要组织相容性复合物（MHC）Ⅱ表达的影响。结果1μg/mlHMGB1刺激48h后,DC表面受体RAGE表达明显上调（P〈0.01）;在1：50、1：100、1：200稀释度的抗RAGE多克隆抗体作用后,HMGB1刺激诱导DC表面CD80、CD86和MHCⅡ表达减弱（P〈0.01）,其中1：100稀释度时表达减弱最明显。结论HMGB1能诱导DC受体RAGE表达增强,RAGE可能是参与HMGB1诱导DC成熟分化的重要受体。
英文摘要：
Objective To investigate the effects of high mobility group box-1 protein （HMGB1） on surface receptor expression of splenic dendritic cells （DCs） and its potential mechanism in rats. Methods DCs isolated from the spleens of male Wistar rats were seeded on 96-well （1 × 10^5 cells/well） cell culture plates, and the cells were stimulated with HMGB1 in a concentration of 1μg/ml for 48h. After being stimulated, DCs were denatured in cell culture plates to determine the expression of the receptor for advanced glycation end products （RAGE） with flow cytometry and laser scanning confocal microscopy. The dose-dependent responses between anti-RAGE polyclone antibody and costimulatory molecules including CEN）, CD86 and major histocompatibility complex （MHC） Ⅱ expressions on the surface of DCs stimulated with HMGB1 were analyzed with flow cytometry. Results After stimulation with 1μg/ml HMGB1 for 48h, it was noted that DCs could be induced into maturation and differentiation with marked expression of CD80, CD86 as well as MHC Ⅱ. Meanwhile, RAGE expression on the surface of rat splenic mature DCs （rnDCs） was markedly up-regulated （all P〈0. 01）. When DCs were cultured in the presence of 1：50, 1：100 and 1：200 dilution RAGE polyclone antibody for 2 hours, expressions of the costimulatory molecules including CDS0, CD86 and MHC Ⅱ were significantly down-regulated （P〈0. 01） after stimulated with HMGB1 in a concentration of 1μg/ml for 48h, and values of these molecules were lowest in 1：100 dilution （P〈0. 01）. Conclusion The present study demonstrated that HMGB1 stimulation can result in marked up-regulation of RAGE expression on the surface of rat splenic DCs, and RAGE appears to be a potential receptor associated with DCs maturation and differentiation induced by HMGB1.
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