miR-145靶向PDCD4在缺血性脑卒中的作用机制  

The Mechanism of miR-145 Targeting PDCD4 in Ischemic Stroke

作　　者：刘宏伟[1] 姚建辉[2] 刘鹏军 李卫民[1] LIU Hong-wei;YAO Jian-hui;LIU Peng-jun;LI Wei-min(Dept.of Emergency;Dept.of Critical Medicine,The First Hospital,Yulin Shaanxi 719000,China)

机构地区：[1]榆林市第一医院急诊科,陕西榆林719000 [2]榆林市第一医院重症医学科,陕西榆林719000

出　　处：《昆明医科大学学报》2020年第4期18-24,共7页Journal of Kunming Medical University

基　　金：陕西省科技厅科研基金资助项目(2013C214)。

摘　　要：目的探讨mi R-145对缺血性脑卒中引起的神经干细胞凋亡和炎性反应的分子机制。方法采用H2O2处理小鼠神经干细胞NE-4C细胞构建缺血性脑卒中细胞模型,流式细胞仪检测NE-4C细胞凋亡水平;RT-q PCR检测mi R-145和炎性相关因子(IL-1β、IL-6、TNF-α)的表达;采用双荧光素酶报告基因验证mi R-145与PDCD4之间的靶向关系;Western blot检测PDCD4的表达水平。结果 H2O2能够引起NE-4C细胞凋亡并上调IL-1β、IL-6、TNF-α的表达(P<0.01或P<0.05)。mi R-145在H2O2处理的NE-4C细胞表达下调;当过表达mi R-145时能显著抑制NE-4C细胞凋亡并下调IL-1β、IL-6、TNF-α的表达(P<0.01或P<0.05)。双荧光素酶报告基因证实mi R-145靶下调PDCD4的表达水平(P<0.01)。实验进一步证实,mi R-145通过靶向下调PDCD4的表达水平抑制NE-4C细胞凋亡并下调IL-1β、IL-6、TNF-α的表达(P<0.01或P<0.05)。结论过表达mi R-145靶向下调PDCD4抑制H2O2处理引起的NE-4C细胞凋亡和炎性反应。Objective To explore the molecular mechanism of mi R-145 on neural stem cell apoptosis and inflammatory response induced by ischemic stroke. Me thods Construction of ischemic stroke cell model by treatment of mouse neural stem cell NE-4 C with H2 O2. Flow cytometry was used to detect the apoptosis level of NE-4 C cells. The expressions of mi R-145 and inflammatory related factors(IL-1β, IL-6,TNF-α) expression levels were detected by RT-qPCR. Dual luciferase reporter assay was applied to verify the targeted regulatory relationship between mi R-145 and PDCD4. The expression level of PDCD4 was detected by Western blot.Re s ults H2 O2 induced apoptosis of NE-4 C cells and up-regulated the expressions of IL-1β,IL-6 and TNF-α (P<0.01 or P <0.05). mi R-145 was down-regulated in H2 O2-treated NE-4 C cells;Overexpression of mi R-145 significantly inhibited NE-4 C cell apoptosis and down-regulated the expression levels of IL-1β,IL-6,TNF-αexpression( P <0.01 or P <0.05). Dual luciferase reporter gene confirmed that mi R-145 targeted down-regulationof PDCD4 expression(P<0.01). Furthermore,we found that mi R-145 inhibited the apoptosis of NE-4 C cells and down-regulated the expression of IL-1β, IL-6 and TNF-α by down-regulating the expression level of PDCD4(P <0.01 or P <0.05). Conclus ion Overexpression of mi R-145 targets down-regulation of PDCD4 to inhibit apoptosis and inflammatory responses in NE-4 C cells induced by H2 O2 treatment.

关 键 词：缺血性脑卒中 细胞凋亡 炎性反应 MIR-145 PDCD4 

分 类 号：R45[医药卫生—治疗学]