荧光定量PCR验证大鼠严重烫伤后早期骨骼肌代谢高通量芯片中miRNA-30a表达及意义.pdfVIP

.822. 临床军医杂志 2016 年8 月第44 卷第8 期 Clin J Med Offic ,Vol. 44 ,No. 8 ， A吨，2016 t仓 毫雪· 荧光定量 PCR 验证大鼠严重烫伤后早期骨髓肌代谢 高通量芯片中 miRNA-30a 表达及意义 张海军， 柴家科， 郝岱峰， 中传安， 郁永辉 解放军总医院第一附属医院全军烧伤研究所烧伤整形科，北京 100037 [摘要] 目的探讨miRNA-30a 在大鼠严重烫伤后早期骨骸肌代谢中的表达及意义。方法 建立严重烫伤大鼠模型，将 16 只大鼠分为实验组与对照组，每组各8 只，于伤后 72h 留取腔骨前肌标本。应用 TRIZOL 法抽提腔骨前肌中总 RNA ，采用 丹麦锁核昔酸技术 Exiqon miRCURyTM LNA microRNA 芯片(第七代) ，约 7∞条捕获探针，数据库版本为 v. 18.0 ，分别检测实 验组与对照组腔骨前肌标本中 miRNA-30a 的表达水平，实时定量聚合酶链反应(PCR) 验证以检验芯片数据的可靠性。 结果 芯片筛查结果显示，miRNA-30a 在大鼠严重烫伤后早期骨髓肌代谢中表达下调，而荧光定量配R 结果同样显示 miR­ NA-30a 表达降低。结论 m刚A拍阳在大鼠严重烫伤后早期骨儒肌代谢过程中表达下调明显，其作用机制需进一步研究证实。 [关键词] 微小 RNA; 大鼠; 烫伤; 骨儒肌; 表达 中图分类号:阳文献标志码:叫doi: 10.166创/j. 1671-3826 却16.08 口|文章编号:1671-3826(2016 )08 Exprωsion and sigI咀ficance of miRNA-30a in skeletal muscle of rats after severe burns by microarray and real-time quanti- tative PCR ZHANG Hai-jun ,CHAI Jia-ke ,HAO Dai-feng ,SHEN Chuan-an ,YU Yong-hui (Department of Bum and Plastic Surge巧， Bum institu- te ,The First Hospital Affiliated to General Hospital of PLA ,Beijing 10∞48 ， China) Abstract: Objective To explore the miRNA-30a expression in skeletal muscle of rats after severe bums by real-time quantitative PCR and analyzed its biological significance. Methods The Danish lock Exiqon miRCURY ( tm) LNA microRNAs oligonucleotide chip was performed( the seventh generation) ,about 700 capture probes ,databases were version v. 18.0 ,detection of tibialis anterior in the experimental group and the control group took the injured after 72 hours( n =4). Total RNA was extracted with Trizol from skeletal muscles ,the expression of miRNA-30a is examined by real-time quantitative polymerase chain reaction( PCR) with Eva-Green staining. Results The microarray chip showed that the expr