Han Xing, Ding Xin, Xu Li-Xiao, Liu Ming-Hua, Feng Xing
Department of Neonatology, Children's Hospital of Soochow University, Suzhou, Jiangsu 215000, China.
Zhongguo Dang Dai Er Ke Za Zhi. 2016 Mar;18(3):270-6. doi: 10.7499/j.issn.1008-8830.2016.03.016.
To study the changes of miRNA expression in the pineal gland of neonatal rats with hypoxic-ischemic brain damage (HIBD) and the possible roles of miRNA in the pathogenesis of circadian rhythm disturbance after HIBD.
Seven-day-old Sprague-Dawley (SD) rats were randomly divided into 2 groups: HIBD and sham-operated. HIBD was induced according to the Rice-Vannucci method. The pineal glands were obtained 24 hours after the HIBD event. The expression profiles of miRNAs were determined using GeneChip technigue and quantitative real-time PCR (RT-PCR). Then the miRNA which was highly expressed was selected. The expression levels of the chosen miRNA were detected in different tissues (lungs, intestines, stomach, kidneys, cerebral cortex, pineal gland). RT-PCR analysis was performed to measure the expression profiles of the chosen miRNA and the targeted gene Clock mRNA in the pineal gland at 0, 24, 48 and 72 hours after HIBD.
miRNA-182 that met the criteria was selected by GeneChip and RT-PCR. miRNA-182 was highly expressed in the pineal gland. Compared with the sham-operated group, the expression of miRNA-182 was significantly up-regulated in the pineal gland at 24 and 48 hours after HIBD (P<0.05). Compared with the sham-operated group, Clock mRNA expression in the HIBD group increased at 0 hour after HIBD, decreased at 48 hours after HIBD and increased at 72 hours after HIBD (P<0.05).
miRNA-182 may be involved in the pathogenesis of circadian rhythm disturbance after HIBD.
研究缺氧缺血性脑损伤(HIBD)新生大鼠松果体中微小RNA(miRNA)表达的变化,以及miRNA在HIBD后昼夜节律紊乱发病机制中的可能作用。
将7日龄的Sprague-Dawley(SD)大鼠随机分为2组:HIBD组和假手术组。按照Rice-Vannucci法诱导HIBD。HIBD造模后24小时获取松果体。采用基因芯片技术和定量实时聚合酶链反应(RT-PCR)测定miRNA的表达谱。然后筛选出高表达的miRNA。检测所选miRNA在不同组织(肺、肠、胃、肾、大脑皮质、松果体)中的表达水平。进行RT-PCR分析,以检测HIBD后0、24、48和72小时松果体中所选miRNA和靶向基因Clock mRNA的表达谱。
通过基因芯片和RT-PCR筛选出符合标准的miRNA-182。miRNA-182在松果体中高表达。与假手术组相比,HIBD后24和48小时松果体中miRNA-182的表达显著上调(P<0.05)。与假手术组相比,HIBD组Clock mRNA表达在HIBD后0小时升高,48小时降低,72小时升高(P<0.05)。
miRNA-182可能参与HIBD后昼夜节律紊乱的发病机制。