中文摘要：

膜联蛋白1（ANXA1）是一种抗炎因子，但是ANXA1对单核细胞来源树突状细胞（monocyte-derived dendritic cells，MoDCs）功能成熟的影响还未见报道。首先以pShuttle-CMV为载体，通过一系列分子克隆技术，构建了ANXA1高表达质粒pShuttle-CMV-ANXA1。笔者将pShuttle-CMV-ANXA1转染细胞株293a，用western-blot法验证ANXA1高表达情况。结果表明，pShuttle-CMV-ANXA1可引起293a表达ANXA1增高；表明，pShuttle-CMV-ANXA1构建是成功的。然后，用脂质体转染试剂GenePorterⅡ将荧光对照质粒pShuttle-CMV-VNGIRES转染TNFα-MoDCs。通过镜下荧光可知，GenePorterⅡ将对照质粒转染MoDCs的效率〉80％；表明，GenePorterⅡ是一种较好的转染MoDCs的方法。用GenePorterⅡ将pShuttle-CMV及pShuttle-CMV-ANXA1转染TNF-α-MoDCs，通过western-blot法检测ANXA1在TNF-α-MODCs中的高表达情况。结果表明，与pShuttle-CMV相比，pShuttle-CMV-ANXA1可引起TNF-α-MoDCs高表达ANXA1；成功将pShuttle-CMV-ANXA1转入MoDCs，并得到表达。GenePorterⅡ转染树突状细胞的新方法，简单高效，为进一步研究ANXA1对MoDCs的生物学功能奠定了基础。获得的高表达ANXA1的树突状细胞，可望在自身免疫病及移植排斥反应中得到应用。

英文摘要：

Although ANXA1 has been reported to work as an anti-inflammatory factor, its role on the maturation of monocye-derived dendritic cells （MoDCs） has not been examined. In this study, we constructed ANXA1 over-expression plasmid and investigated the method for transfecting MoDCs. Firstly, ANXA1 over-expressing plasmid was constructed by a series of molecular cloning technigues. Then, the over-expression plasmid was transfected into 293a cell line and the ANXA1 expression was detected by Western-Blot. Compared with control plasmid, ANXA1 expression was up-regulated in 293a cell line which was transfected with over-expression plasmid. The results suggested that ANXA1 over-expressing plasmid was constructed successfully. Secondly, the fluorescent control plasmid was transfected into MoDCs using transfection reagent, GenePorter Ⅱ. The transfection efficacy under fluorescent microscope was more than 80 %. The results suggestsed that GenePorter Ⅱ was a useful reagent for MoDCs transfection. Finally, the plasmids, pShuttle-CMV and pShuttle-CMV-ANXA1 were transfected into TNF-α- MoDCs and the ANXA1 expression was detected by Western-Blot. The results showed that compared with pShuttle-CMV, ANXA1 was up-regulated in TNF-α-MoDCs which was transfected with pShuttle- CMV-ANXA1. The results suggested that ANXA1 was successfully transfected into and over-expresssed in TNF-α-MoDCs. The easy and effective method to transfect dendritic cell provides a basis for the further biological study of ANXA1 on MoDCs. The dendritic cells over-expressing ANXA1 could be used in the treatment of autoimmune disease and transplant reiection.