miR-1187靶向调控Arhgef9在七氟醚致海马神经元凋亡中的作用  

Regulatory effect of miR-1187 on Arhgef9-mediated apoptosis of hippocampal neuron in mice

作　　者：孙华琴 梁哲浩[2] 徐孝平[3] 胡弘毅[1] 陶涛[1] UN Huaqin;LIANG Zehao;XU Xiaoping(Department of Anesthesiology, the First Affiliated Hospital of Zhejiang Chinese Medical University, Hangzhou 310006, China)

机构地区：[1]浙江中医药大学附属第一医院麻醉科,杭州310006 [2]浙江中医药大学附属第一医院超声诊断科,杭州310006 [3]浙江中医药大学动物实验研究中心比较医学研究所

出　　处：《浙江医学》2019年第10期991-994,I0005,共5页Zhejiang Medical Journal

基　　金：浙江省医药卫生科技计划(2014KYA159);浙江省中医药科技计划(2012ZA048;2019ZA047)

摘　　要：目的探讨miR-1187靶向调控Rho鸟苷交换因子-9(Arhgef9)在七氟醚致小鼠海马神经元凋亡中的调控作用。方法通过基因芯片技术测定小鼠海马miRNA。利用miR Walk数据库对差异miRNA的靶基因进行预测。为验证预测结果与实际情况是否相符,同步进行mRNA水平的表达谱芯片实验,筛选出与差异miRNA确有互作关系的m RNA。为说明miRNA与m RNA的互作关系,进一步利用David数据库对靶基因进行注释,最终形成miRNA与m RNA共表达网络。采用RT-PCR法检测海马神经元miR-1187mRNA相对表达量,细胞计数试剂盒(CCK-8)法检测miR-1187转染后海马神经元存活率,荧光素酶实验检测Arhgef9是否为miR-1187的靶基因。结果海马神经元凋亡主要与9个miRNA相关,即miR-101b-3p、miR-1187、miR-188-5p、miR-219a-5p、miR-338-3p、miR-425-5p、miR-467a-3p、miR-705、miR-92a-3p。miR-1187在七氟醚诱导海马神经元损伤模型中的相对表达量明显降低(P<0.05),miR-1187高表达质粒(miR-1187 mimics)转染后,海马神经元存活率明显升高(P<0.05)。miR-1187与Arhgef9存在靶向关系,miR-1187通过与Arhgef9基因m RNA的3′非翻译区互补配对来发挥作用。结论 miR-1187是海马神经元抗细胞凋亡的抑制因子;高表达miR-1187通过靶向调节Arhgef9 m RNA表达来抑制海马神经元凋亡的发生。Objective To investigate the regulative effect of miR-1187 on Arhgef9-mediated apoptosis of hippocampal neurons in mice. Methods Mouse hippocampal microRNA was determined by gene chip technology. Target genes for differential miRNAs were predicted using the miR Walk database. The mRNAs interacting with the differential miRNAs were screened with microarray to verify the predicted results. The interaction between miRNA and mRNA was annotated with the David database to form a co-expression network of miRNAs and mRNAs. The expression of miR-1187 mRNA was detected by RT-PCR. The survival rate of hippocampus cells after miR-1187 transfection was detected by CCK-8. The Arhgef9 was detected with luciferase reporter technique to be proved as a target gene of miR-1187. Results Hippocampal neuronal apoptosis was associated with miR-101b-3p, miR-1187, miR-188-5p, miR-219a-5p, miR-338-3p, miR-425-5p, miR-467a -3p, miR-705 and miR-92a-3p. The expression of miR-1187 was significantly decreased in the model of hippocampal cell injury induced by sevoflurane (P<0.05). After miR-1187 mimics transfection, the survival rate of hippocampus neurons was significantly increased (P<0.05). There was a targeting relationship between miR-1187 and Arhgef9, and miR-1187 played its role by complementary pairing with the 3'UTR of Arhgef9 gene mRNA. Conclusion High expression of miR-1187 inhibits the apoptosis of hippocampal neurons by targeting the regulation of Arhgef9 mRNA expression in mice.

关 键 词：miR-1187 RHO 鸟苷交换因子-9 海马神经元 细胞凋亡 

分 类 号：R614[医药卫生—麻醉学]