【摘要】 于上海地区汉人群中,探讨阿尔茨海默病(AD)患者外周血中β淀粉样前体蛋白基因外显子9、10(APP9～10)的mRNA表达水平,并作cDNA的点突变检测。采用半定量竞争性反转录聚合酶链反应(semi quantitivecompetitiveRT PCR)技术测定APP9～10的mRNA表达水平;以聚合酶链反应(PCR)-限制性片段长度多态(RFLP)方法检测基因组DNA中载脂蛋白E基因(APOE)和早老素1基因(PS1)多态;APP9～10cDNA的点突变检测采用变性梯度凝胶电泳法。结果表明(1)AD患者中APP9～10的mRNA表达水平显著高于健康老人(P<0.05);(2)作为AD的主要风险因子,APOE ε4等位基因并不影响AD患者中APP9～10的mRNA表达水平,而另一可疑风险因子PS1基因1等位基因的存在却可显著增加AD患者中APP9～10的mRNA表达;(3)AD患者中不存在APP9～10cDNA的点突变。提示上海汉人群中,AD患者外周血中APP基因外显子9、10的mRNA表达水平显著升高,也许这一生物学改变可作为一种生物学辅助诊断指标应用于AD的临床研究。更多还原

【Abstract】 To explore the expression differences of exon 9 and 10 in Amyloid Precursor Protein gene(APP9～10) in Alzheimer′s disease,and detect the probable point mutation appeared in cDNA fragment of APP9～10 in the Shanghai Han people.semiquantitaive competitive RTPCR technique was performed to detect the expression of APP9～10 in peripheral lymphocyte,and the Apolipoprotein E gene(ApoE) and Presenilin 1(PS1)gene were genotyped with PCRRFLP method.We also analyzed the point mutation in APP9～10 cDNA through the denatured gel eletrophoresis.The results are as follows:1.While compared with healthy controls,expression of APP9～10 mRNA was significantly enhanced in Alzheimer disease;2.APOE*ε4 allele,the most common genetic risk factor for AD,did not affect the Expression of APP9～10 mRNA,whereas the APP9～10 mRNA expression might be increased by the allele 1 of PS1 gene,another probable susceptibility gene of AD.3.No point mutation in APP9～10 cDNA was detected.In our samples,the expression of APP9～10 mRNA in AD was significantly different from that of controls,suggesting that the change of peripheral APP9～10 mRNA expression might be another biomarker used in clinical diagonosis for AD.更多还原