题名页 micrornas荧光定量pcr检测方法的建立及其在急性肺损伤中 .doc

题名页才志刚、张绍明、张珩、周宜勇、吴海波、徐小平通讯作者：才志刚 地址：上海市淮海西路338号解放军455医院 电话：021传真：021-mail: caizg12345@ 总页数：图数：文章字数： This work was supported by the Medical Science Foudation of Nanjing Military Command (No. 09MA028). 本研究工作受2009年南军区医学科研立项课题基金资助（No.09MA028）。 Corresponding author. Tel: +86-21 Fax: +86-21 E-mail: caizg12345@ MicroRNAs荧光定量PCR检测方法的建立及其在急性肺损伤中的表达分析 才志刚*、张绍明、张珩、周宜勇、吴海波、徐小平200052 摘要：MicroRNA (miRNA) 是发现一类具有转录后调节活性的小分子RNA转录后负性调节蛋白质表达miRNA参与了许多重要的细胞Department of Cardio-Thoracic Surgery, Number 455 Hospital of The Chinese People’s Liberation Army, Shanghai 200052, China Abstract: microRNA (miRNA) was a class of highly conserved, single strand, non-coding small RNA that can regulate the expression of target mRNAs post-transcriptionally either through translational inhibition or destabilization of target mRNAs. A novel miRNA quantification method has been developed using stem-loop RT followed by real time PCR analysis. To investigate the expression level of miRNAs in acute lung injury, we established the miRNA quantification method using Real-time PCR. Verification of the dynamic range and melt curve of qRT-PCR indicated that the method we established here showed highly specificity and precision. By analysis of the expression of miR-1, an muscle-specific miRNA, using our quantification method, we showed, in the four tissues examined here, miR-1 was only detected in heart, suggesting our method is sensitivity and specifity to observe the tissue miRNA expression level. To gain insights into the expression profile of miRNAs in acute lung injury, mouse lung injury model was established using LPS intratracheal administration. Using real-time PCR analysis , we found that a collection of miRNAs was dynamically regulated in LPS induced lung injury, indicating that miRNAs played an important role in the processes of acute lung injury. Key words: acute lung injury; microRNA; stem-loop; PCR MiRNA是在果蝇、线虫、小鼠和人等多种生命体中发现一类具有转录后调节活性的小分子RNA其它一起形成了RNA诱导的沉