红景天苷干预人胚肺二倍体成纤维细胞衰老的microRNA芯片筛选与分析  

The miRNA microarray analysis of salidroside's anti-aging function in human embryonic lung fibroblast 2BS cells

作　　者：张婧[1] 苏慧丽[1] 王雅珍[1] 文晓林[1] 贾兵兵[1] 毛根祥[1] 

机构地区：[1]浙江医院,浙江杭州310013

出　　处：《药学学报》2017年第4期550-555,共6页Acta Pharmaceutica Sinica

基　　金：国家自然科学基金资助项目(31201040);浙江省医药卫生平台计划(学科带头人)(2015DTA001);国家卫计委科学研究基金项目(WKJ2011-2-014);浙江省中医药科学研究基金项目(2015ZA002);浙江省卫生医药科研项目(2016KYB005)

摘　　要：本文运用microRNA芯片方法检测红景天苷干预人胚肺二倍体成纤维细胞衰老过程中的microRNA表达变化,并预测miRNA调控的靶基因功能与信号通路。将人胚肺二倍体成纤维细胞分为3组:年轻细胞(28 PD,young)、近复制衰老(50 PD,old)和红景天苷处理的50 PD细胞(old+SAL),进行miRNA表达谱分析。结果找到43个miRNA与细胞复制性衰老显著相关。58个miRNA在红景天苷干预后表达发生了改变。6个miRNA分子包括hsa-let-7c、hsa-let-7e和hsa-mir-3620表达量在衰老细胞组中表达降低而在红景天苷干预后表达增加,hsa-mir-411、hsa-mir-24-2-5p和hsa-mir-485-3p则表现出相反的趋势。对已经报道过的靶基因进行功能聚类和信号通路分析,靶基因功能明显富集于31个GO,11条信号通路。挑选4个miRNA经RT-PCR验证结果与芯片结果相符。本研究结果为从microRNA水平探讨红景天苷抗衰老机制提供了线索。This study was designed to investigate the microRNA expression profile in human embryonic lung fibroblast 2BS cells upon salidroside （SAL） treatment, and predict the target genes of miRNAs and related pathways delaying cellular senescence. Samples were divided into three groups： young control （28 PD）, old control （50 PD）, and old＋SAL （50 PD with SAL）, RNA from three groups was used for miRNA microarray analysis. In late PD cells, 43 miRNAs were found significantly changed relatively to those in young cells, and 58 miRNAs were regulated by SAL. The miRNAs including hsa-let-7c, hsa-let-7e and hsa-mir-3620 were significantly down-regulated in late PD cells which could be reversed by SAL treatment. However, hsa-mir-411, hsa-mir-24-2-5p and hsa-mir-485-3p exhibited an opposite trend. Gene Ontology and Pathway analysis revealed that target genes were significantly enriched in 31 GO and 11 pathways. The microarray data was further validated with qRT-PCR. This research provides new clues regarding the underlying mechanisms of SAL on cellular senescence through miRNAs regulation.

关 键 词：红景天苷 衰老 MIRNA芯片 成纤维细胞 LET-7 

分 类 号：R966[医药卫生—药理学]