This study was purposed to identify endothelial nitric oxide synthase (eNOS) mRNA in human RBCs during storage and to investigate the relationship of its changing profile and preservation time at 4℃. RT-PCR and gene sequencing were applied to identify eNOS-mRNA in banked RBC. Real time PCR was used to study the relationship of eNOS-mRNA expression and preservation time. The results showed that eNOS mRNA was detected in RBC. Compared with fresh RBC, the content of eNOS mRNA in RBC was 0.868 ±0.119 stored for 1 week, which was 0.379 ±0.289, 0.108 ±0.134, 0.141 ±0.141, 0.125 ±0.12 stored for 2, 3, 4 and 5 weeks respectively. It is concluded that eNOS mRNA exists in human RBC and its content is decreasing gradually along with the prolongation of storag time in banked RBC. Stored for 3 weeks, the content of eNOS-mRNA remains to be at lower level of concentration in human RBC.%本研究鉴定红细胞中有无内皮型一氧化氮合成酶(eNOS) mRNA,探讨在4℃保存条件下红细胞中的eNOS mRNA随保存时间变化的规律.采用逆转录聚合酶链反应(RT-PCR)和基因测序方法检测库存红细胞中有无eNOS mRNA的表达,采用实时荧光定量-聚合酶链反应(real time PCR)检测库存红细胞中eNOS mRNA的表达随时间变化的规律.结果表明:红细胞中eNOS mRNA检测结果呈阳性;保存1周的红细胞中eNOS mRNA含量为新鲜红细胞的0.868±0.119,保存2周为0.379±0.289,3周为0.108±0.134,4周为0.141 ±0.141,5周为0.125±0.12.结论:在红细胞中检测到eNOS mRNA;随着保存时间延长红细胞中eNOS mRNA含量逐渐下降,保存3周后eNOS mRNA表达量维持在一个低浓度水平.
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