文献类型：期刊文献

英文题名：Identification of stable reference genes in plasma astrocyte-derived exosomal mRNA for RT-qPCR analysis in Alzheimer's disease

作者：Guo, Guigui[1];Qu, Yiwen[1];Wang, Qiyao[1];Xiao, Zhihua[2];Liu, Qin[1];Zeng, Xianfeng[3,4];Chen, Shouwen[1]

机构：[1]East China Univ Sci & Technol, Shanghai Frontiers Sci Ctr Optogenet Tech Cell Met, State Key Lab Bioreactor Engn, Shanghai 200237, Peoples R China;[2]Shanghai OPM Biosci Co Ltd, Shanghai 201321, Peoples R China;[3]Yanan Univ, Dept Lab Med, Daxing Hosp, Xian 710000, Peoples R China;[4]Northwest Univ, Sch Med, Xian 710069, Peoples R China

年份：2026

卷号：814

外文期刊名：BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS

收录：;WOS:【SCI-EXPANDED(收录号:WOS:001723883000001)】；

基金：The study was conducted in accordance with the Declaration of Helsinki, and approved by Ruijin Hospital (protocol code: 2022-254) . The presented work was supported by Science and Technology Commission of Shanghai Municipality (24HC2820500) , the National Key Research and Development Program of China (2022YFC3400103) , and the ECUST-OPM Open Fund (20220701) .

语种：英文

外文关键词：Reference gene; RT-qPCR; Astrocyte-derived exosome; mRNA; Alzheimer's disease

摘要：Accurate quantification of relative gene expression by RT-qPCR requires the selection of appropriate reference genes for data normalization. Recently, extracellular vesicle components, particularly exosomal RNAs, have recently emerged as promising biomarkers for Alzheimer's disease (AD). Among these, mRNAs from astrocytederived exosomes (ADEs) hold particular significance for AD diagnosis and prognosis due to their high in vivo stability and capacity to cross the blood-brain barrier, thereby faithfully reflecting cerebral pathological changes in AD patients. However, suitable reference genes for normalizing target gene expression in ADEs have not been systematically identified. In this study, we comprehensively evaluated the expression stability of candidate reference genes in plasma ADEs across cognitively unimpaired (CU), mild cognitive impairment (MCI), and AD patients using five robust algorithms, including Genorm, Bestkeeper, Delta Ct method, Normfinder, and RefFinder. Our results revealed that YWHAE and RNPS1 exhibited superior expression stability across all three groups, whereas GAPDH, a conventionally used reference gene, demonstrated inadequate stability. Collectively, our work provides the first systematic validation of reference genes for mRNA quantification in plasma ADEs, establishing a methodological foundation for ADE-based AD biomarker development.