【摘要】 目的探寻储存红细胞添加一氧化氮(NO)供体硝普钠(SNP)后红细胞内特定microRNA表达水平的变化,以了解体外添加NO对与红细胞衰老、凋亡相关的microRNAs表达的影响。方法 3(人)份血样来自健康无偿献血者血液制备的悬浮红细胞,滤除白细胞后均分为4℃储存20 d红细胞组(储存组)和4℃储存20 d红细胞添加1μmol/L SNP组(SNP组);对2组标本用基因芯片检测,根据差异倍数≥2或P≤0.05 2个条件,以火山图筛选出差异基因,并结合已有文献报道从中筛选出与细胞衰老、凋亡相关的microRNAs作RT-qPCR验证。结果基因芯片检测:SNP组与储存组比较,发现有69个上调基因和52个下调基因;RT-qPCR验证试验:筛选出与细胞衰老、凋亡相关的13个microRNAs没有明显变化(P>0.05)。结论体外添加NO供体SNP后储存红细胞内microRNAs的基因表达有变化,但NO短期内不影响与红细胞衰老、凋亡相关的microRNAs表达,显示其对红细胞的寿命可能无影响。更多还原

【Abstract】 Objective To investigate the effects of addition nitric oxide donor—sodium nitroprusside(SNP) on the expression of apoptosis-related miRNAs in stored RBCs in suspension.Methods Three blood samples were collected from healthy donors and made into suspended red cells.The leukocytes were filtrated,then divided into 20 days stored suspension RBCs and 20 days stored suspension RBCs incubated with 1 μmol/L SNP.RNA were isolated and subjected to miRNA profiling by using microarray,then differential expression genes were isolated according to the fold-change ≥2 or P≤0.05.13 miRNAs were chosen by Volcano Plot and the references.miRNAs were verified by RT-q PCR.Results MiRNA profile results showed that 69 up-regulated miRNAs and 52 down-regulated miRNAs were found when SNP was added to stored RBCs.SNP incubation did not affect the expression of selected apoptosis associated miRNAs(P>0.05).Conclusion Microarray data show miRNAs expression changes in stored RBC after SNP addition,but no impact on the expression of miRNAs related to cell senescence and apoptosis.These results suggest that NO does not affect the RBCs lifespan.更多还原