七氟烷麻醉大鼠海马组织特异microRNA的差异表达及靶标预测
Differential expression and target prediction of microRNA in hippocampus in Sevoflurane-anesthetized rats
杨宇帆 1潘波 1李思源 1黄金勇 1刘驰 2潘志强1
作者信息
- 1. 徐州医学院麻醉学院,江苏徐州221004
- 2. 徐州医学院临床医学系,江苏徐州221004
- 折叠
摘要
目的 定量检测和分析七氟烷麻醉SD大鼠海马miR-133a、miR-96、miR-183的表达,并结合生物信息学,分析预测差异表达miRNA的调控靶标基因.方法 54只SD雄性大鼠以6L/min通气量不同浓度七氟烷(1.5％、2.5％、3.5％)麻醉4h,苏醒后不同时间(0 h、4 h、2 d和10 d,每组6只)提取大鼠海马miRNA,并反转录为cDNA,用RT-qPCR方法对miRNA进行定量分析.使用3种生物信息学预测软件对miR-133a、miR-96、miR-183进行靶标分析.结果 miR-133a、miR-96、miR-183在麻醉后大鼠海马中均有表达,但表达差异不一致,与未麻醉的大鼠相比,miR-133a差异最显著,麻醉后0 h、4 h、2 d和10d分别下调2、1、2.5倍和1倍.信息学分析表明,mirSV score在1.0以上得分有12个靶标基因,其中3个靶基因与记忆障碍有关,分别为溶质载体家庭6成员1 (Slc6a1)、腺苷酸环化酶(Adcyap1)和蛋白磷酸酶催化亚基(Ppp2c),它们可能与七氟烷麻醉导致的记忆减退相关.结论 miR-133a可能参与七氟烷致学习记忆下降相关,它可能通过多个基因参与对麻醉后记忆障碍的调控.
Abstract
Objective To quantitatively analyze the differential expression of miR-133a,miR-96,miR-183 in hippocampus of Sevoflurane-anesthetized rats,combined with the bioinformatics predicte to the potential targets of the differential expressed miRNAs.Methods 54 male SD-rats were anesthezied for 4 h by the use of 6 L/min ventilatory capacity different concentrations of Sevoflurane including 1.5％,2.5％ and 3.5％.Rats hippocampus was harvested after different aweaking time (0 h,4 h,2 d and 10 d).miRNAs were extracted from hippocampus and reverse transcript into cDNA.Then,RT-qPCR was used to detect and analyze the expression of these miRNAs.Three bioformatics sofiwares including Target,PicTar and Micro were utilized to predict the miRNA target.Results Three miRNA including miR-133a,miR-9 and miR-183 were all expressed in hippocampus of Sevoflurane-anesthetized rats.However,their differential values were inconsistent.Of three miNRAs from Sevoflurane-anesthetized rats,miR-133a was most differentially expressed compared with unanesthetized-rats,and had 2 fold differential down-regulation for 0 h aweaking,1 fold for 4 h aweaking,2.5 fold for 2 d aweaking and 1 fold for 10 d after anesthetization,respectively.The analysis of bioinformatics indicated there were 12 targets for miR-133a.Among these targets,Slc6a1,Adcyap1 and Ppp2c were possibly associated with memory impairment induced by Sevoflurane.Conslusion miR-133a may be implicated in memory impairment,which may be regulated by several genes together.
关键词
miRNA/七氟烷/学习记忆障碍Key words
miRNA/Sevoflurane/Learning and memory impairment引用本文复制引用
基金项目
出版年
2013
NETL
NSTL
维普
万方数据