抗自身免疫的新途径-CⅡTA siRNA的体外筛选及活性  

In vitro biological activity of CⅡTA siRNA:a novel approach for the treatment of autoimmune disease

作　　者：郭荣[1] 何飞[2] 杜欣[1] 罗成伟[1] 陆泽生[1] 翁建宇[1] 吴穗晶[1] 林伟[1] 

机构地区：[1]广东省人民医院血液科,广州510080 [2]郑州大学第一附属医院心内科,郑州450052

出　　处：《免疫学杂志》2008年第3期264-267,共4页Immunological Journal

基　　金：广东省医学科学技术研究基金资助(A2005017);广东省自然科学基金资助(04003959);广东省人民医院科学技术研究基金(Y2004087)资助项目

摘　　要：目的探讨主要组织相容性复合物Ⅱ类转录激活因子(CⅡTA)的siRNA对软骨细胞表面主要组织相容性复合物(MHC)Ⅱ类分子表达的抑制。方法设计并合成针对CⅡTA编码区第176、741及3061位点的siRNA(分别为siRNA1、siR-NA2、siRNA3)。通过纳米载体介导siRNA稳定转染原代软骨细胞,流式细胞术检测经典的MHC-Ⅱ(HLA-DR、-DP、-DQ)类抗原表达,RT-PCR检测CⅡTA及其调控的MHC-Ⅱ类分子的mRNA水平。结果在重组人干扰素(interferon,IFN)-γ诱导下,siRNA3阳性软骨细胞表面HLA-DR、-DP抗原表达分别降低了93.36%、94.16%,同时CⅡTA的mRNA含量明显减少。结论siRNA3抑制了CⅡTA的mRNA含量,并阻止其调控的MHC-Ⅱ类分子的表达。Objective To investigate the inhibitory effects of siRNA against the wansactivator of MHC-Ⅱ molecules （CⅡTA） on the expression of MHC-Ⅱ in chondrecytes. Methods Three siRNAs recognizing the 176, 741, 3 061 site of CⅡTA were respectively synthesized （siRNA1, siRNA2. and siRNA3）. Stable transfectants of chondrocytes with siRNA were analyzed for classic MHC-Ⅱ （HLA-DR,-DP,-DQ） expression by flow cytometry. The level of CⅡTA mRNA was detected by RT-PCR. Results The expressions of HLA-DR,-DP on siRNA3 positive chondrecytes under IFN-γ induction were decreased significantly （93.36% and 94.16% respectively, P 〈 0.05） ; so did the content of CⅡTA mRNA （89.11% ±7.81%, P〈0.05）. Conclusion siRNA3 significantly decreases the content of CⅡTA mRNA, thus blocks the expression of the family of MHC-Ⅱ molecules, which would benefit the development of a novel nucleic acid drug for autoimmune disease.

关 键 词：小干扰RNA MHCⅡ类分子转录激活因子 自身免疫性疾病 

分 类 号：R593.2[医药卫生—内科学]