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

Expression and significance of miRNA-30a in skeletal muscle of rats after severe burns by microarray and real-time quantitative PCR

目的探讨miRNA-30a在大鼠严重烫伤后早期骨骼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72 h留取胫骨前肌标本。应用TRIZOL法抽提胫骨前肌中总RNA,采用丹麦锁核苷酸技术Exiqon miRCURYTMLNA microRNA芯片(第七代),约700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胫骨前肌标本中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骼肌代谢中表达下调,而荧光定量PCR结果同样显示miRNA-30a表达降低。结论 miRNA-30a在大鼠严重烫伤后早期骨骼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。

Objective To explore the miRNA-30 a expression in skeletal muscle of rats after severe burns 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-30 a is examined by real-time quantitative polymerase chain reaction（ PCR） with Eva-Green staining.Results The microarray chip showed that the expression of miRNA-30 a was decreased in burned rats. Real-time PCR showed the reduced expression of miRNA-30 a in the experimental group compared with the control group（ P〈0. 05）. Conclusion The expression of miRNA-30 a was down-regulated in skeletal muscle of rats after severe burns,the mechanism,maybe associated with autophagy,need to be further research.