中文摘要：

目的运用RNA干扰（RNAi）技术研究OIf-1／EBF相关锌指蛋白（OAZ）对其信号通路相关基因的干预作用及对狼疮抗核抗体（ANA）产生的影响。方法采集并分离系统性红斑狼疮（SLE）患者外周血单个核细胞，分为实验组、阳性对照组、阴性对照组和空白对照组进行细胞培养。实验组和阳性对照组分别用OAZ和甘油醛-3-磷酸脱氢酶（GAPDH）小干扰RNA（siRNA）在体外对培养细胞进行干扰，阴性对照组采用无关RNA序列，空白对照组为培养基对照。72h后分离细胞和上清液，从细胞中提取RNA，逆转录为cDNA，用实时荧光定量聚合酶链反应（PCR）分析不同分组OAZ信号通路相关基因OAZ、分化抑制因子（Id）1、Id2、Id3、Id4mRNA表达水平的变化。酶联免疫吸附试验（ELISA）法测定上清液中干扰素（IFN）-γ、白细胞介素（IL）-4、IL-10、IL-12、IL-21、CC趋化因子配体（CCL）2、抗核抗体（ANA）浓度，并分析这些因子分泌水平与OAZ基因表达的相关性。结果实验组OAZ、Id1、Id2、Id3基因的mRNA表达水平（△Ct分别为12．5±1．4、8．9±1．5、4．3±0．8、8．0±1．1）较阴性对照组（△Ct分别为10,2±1．1、6．5±1．2、2．4±1.3、6．2±1．2）显著降低（P〈0．05）；Id4表达水平虽较阴性对照组有低表达趋势，但差异无统计学意义。实验组IFN-1、IL-10、IL-12、IL-21、ANA的水平明显低于阴性对照组；而CCL2高于阴性对照组（P〈0．05）。实验组与阴性对照组OAZmRNA表达水平△△Ct值与ANA的A比值、IL-21浓度比值呈负相关；与Th1／Th2、CCL2比值呈正相关。结论OAZsiRNA可有效降低狼疮外周血单个核细胞中OAZ信号通路相关基因表达，同时减低多种细胞因子及抗核抗体水平，推测OAZ可能通过Id基因影响细胞因子的水平并导致ANA异常表达。

英文摘要：

Objective To explore the role of OAZ gene in the pathogenesis of systemic lupus erythematosus （SLE） by using RNA interfering technique. Methods Peripheral blood mononuelear cells （PBMC） from SLE patients were collected. Each sample was equally divided into four groups for cell culture in 96 well plates. Specific siRNA for OAZ and GAPDH were concordantly added to the experimental group and the positive control group, while nonspeeific siRNA was added to the negative control group and only culture medium was added to the Mock control group. Cells and supernatants were harvested after culturing for 72 hours, then RNA was extracted and reverse transeripted to cDNA. OAZ, Idl, Id2, Id3, ld4 mRNA expression levels were analyzed by using real-time PCR. Levels of IFN-γ IL-4, IL-10, IL-12, IL-21, CCL2, ANA in the supernatant were tested by ELISA. Relationships between the expression levels of OAZ mRNA with levels of cytokines and ANA were analyzed. Results OAZ, Idl, Id2, Id3 gene mRNA expression levels （△Ct： 12.5± 1.4, 8.9±1.5, 4.3±0.8, 8.0±1.1） in the experimental group were significantly decreased comparing to those in the negative control group （△Ct： 10.2±1.1, 6.5±1.2, 2.4±1.3, 6.2±1.2 respectively, P〈0.05）. Levels of IFN-γ IL-10, IL-12, IL-21 and ANA in the experimental group were significantly lower than those in the negative control group （P〈 0.05 ）, but level of CCL2 was higher than the negative control group （P〈0.05）. Difference of OAZ mRNA expression levels （△△Ct） between the experimental group and the negative control group was negatively correlated with changes of ANA, IL-21 levels, but positively correlated with changes of Th1/Th2, CCL2. Conclusion OAZ siRNA can effectively reduce the expression of genes involved in the OAZ signaling pathway in SLE. OAZ may lead to abnormal production of ANA via regulating Id genes and eytokines.