Alzheimer病患者外周血HO-1基因表达水平的改变  

Expression of HO-1 gene in peripheral blood of the patients with Alzheimer′s disease detected by fluorescence quantitative RT-PCR method

作　　者：陈辉 刘明杰 汪华侨[3] 程刚[4] 何蕴绍[4] 

机构地区：[1]山东省烟台山医院生殖中心,山东烟台264000 [2]烟台出入境检验检疫局 [3]中山大学中山医学院人体解剖与组织胚胎学系 [4]中山大学达安基因诊断中心

出　　处：《中国老年学杂志》2006年第4期429-432,共4页Chinese Journal of Gerontology

基　　金：国家计划委员会项目"荧光探针定量诊断试剂盒的研制与开发"子项目(国经贸委1998-345);广东省自然科学基金(010698)

摘　　要：目的建立荧光定量PCR方法检测A lzheim er病(AD)患者外周血中血红素加氧酶1(HO-1)基因的表达水平,并探讨其表达水平的改变与AD发病的关系。方法在HO-1的基因高保守区设计并合成特异性引物和荧光标记探针。将PCR扩增目的片段用AT克隆的方法克隆入T载体,重组质粒经筛选、鉴定后,作为阳性模板,用于标准曲线的制备和样品检测,并用该方法检测30例AD患者和23例正常老年人对照组外周血HO-1基因mRNA水平。结果应用重组质粒制作的定量曲线循环阈值与模板浓度具有良好的线性关系。AD组HO-1基因平均表达水平(7.04×105±5.96×105)cop ies/μg RNA高于对照组(1.62×105±3.31×105)cop ies/μg RNA(P<0.001)。结论建立了荧光定量PCR方法用于检测AD患者外周血HO-1基因的表达水平,具有较好的检测灵敏度和重复性。HO-1在AD患者外周血中的表达有所增加。Objective To establish fluorogenic quantitative PCR method for detecting heme oxygenase-1 （HO-1）expressions in the peripheral blood of patients with Alzheimer＇s disease（AD） ,and to explore relationship between HO-1 levels and AD. Methods According to the specific sequence of HO-1 genes, the primers and the fluorogenic probe were designed and synthesized. The fragments generated from PCR as templates were cloned into the T vector with the method of AT clone. The positive recombinant plasmid was used as standard quantitative template to make the standard curve for sample detection to detect HO-1 mRNA levels in peripheral blood between 30 cases of patients with AD and 23 normal elder controls. Results The standard curve indicated the linear relationship between cycle threshold （CT） and template concentration. The expression levels of HO-1 in AD patients [（7.04×10^5±5.96×10^5）copies/μg RNA] were higher than that in controls [ （1.62×10^5±3.31×10^5）copies/μg RNA] （P 〈 0. 001 ）. Conelusions The contructed method of fluorogenic quantitative PCR to detect the gene expression of HO-1 in AD patients has better sensitivity and repeatability. The expression level of HO-1 in AD patients was increased.

关 键 词：荧光定量PCR ALZHEIMER病 血红素加氧酶1 基因表达 

分 类 号：R749.16[医药卫生—神经病学与精神病学] R378.61[医药卫生—临床医学]