Mir-215-5p induces autophagy by targeting PI3K and activating ROS-mediated MAPK pathways in cardiomyocytes of chicken
Mir-215-5p induces autophagy by targeting PI3K and activating ROS-mediated MAPK pathways in cardiomyocytes of chicken
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Mir-215-5p 通过靶向 PI3K 并激活鸡心肌细胞中 ROS 介导的 MAPK 途径诱导自噬
DOI：
10.1016/j.jinorgbio.2019.01.010
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发表时间：
2019-04-01
影响因子：
3.9
通讯作者：
Zhang, Ziwei
中科院分区：
文献类型：
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作者：
Cai, Jingzeng;Yang, Jie;Zhang, Ziwei
我们先前的研究表明，硒缺乏可通过触发自噬而导致心肌损伤。MicroRNAs(MiRNAs)在自噬细胞死亡中起着至关重要的作用。然而，miRNAs与缺硒所致心肌自噬损伤的关系尚不清楚。我们利用高通量miRNA测序技术在缺硒心肌组织中筛选出差异microRNA-215-5p(miR-215-5p)。为进一步探讨miR-215-5p在心肌损伤中的作用，采用miRNAs干扰技术建立了miR-215-5p在原代心肌细胞模型中的过表达/下调。在本研究中，我们报道miR-215-5p通过直接与磷脂酰肌醇-4，5-二磷酸3-激酶(PI3K)的3‘非翻译区(3’UTR)结合来促进心肌自噬。应用双荧光素酶报告基因分析、实时定量聚合酶链式反应(qRT-PCR)和Western印迹等方法对其靶基因PI3K进行了鉴定。我们的结果表明miR-215-5p的过表达可能通过PI3K-苏氨酸蛋白激酶(AKT)-雷帕霉素(TOR)途径的靶标触发心肌自噬。进一步的研究表明，自噬细胞的死亡依赖于细胞外信号调节的激动素/2(ERK1/2)、c-jun氨基末端激酶(JNK)、p38激酶(P38)的激活以及miR-215-5p过表达所产生的活性氧(ROS)。相反，miR-215-5p抑制剂可以通过抑制Ros丝裂原活化蛋白激酶(MAPK)通路和激活PI3K/AKT/TOR通路来增强细胞抵抗自噬的生存能力。总之，我们的发现支持miR-215-5p可能通过调节自噬来调节细胞存活计划，miR-215-5p在调控反馈环路中扮演自噬调节者的角色，通过调节PI3K/AKT/TOR通路和ROS依赖的MAPK通路来调节心肌细胞的存活。
Our previous study revealed that selenium (Se) deficiency can cause myocardial injury through triggering autophagy. MicroRNAs (miRNAs) play crucial roles in autophagic cell death. However, the relationship between miRNAs and myocardial autophagy injury caused by Se deficiency remains unclear. We selected differential microRNA-215-5p (miR-215-5p) in Se-deficient myocardial tissue using high-throughput miRNA-sequencing. To further explore the role of miR-215-5p in myocardial injury, overexpression/knockdown of miR-215-5p in primary cardiomyocyte model was established by miRNAs interference technology. In this study, we report that miR-215-5p can promote myocardial autophagy by directly binding to the 3'untranslated region (3'UTR) of phosphatidylinositol-4, 5-bisphosphate 3-kinase (PI3K). Its target gene PI3K was confirmed by dual luciferase reporter assay, quantitative real-time polymerase chain reaction (qRT-PCR) and western blot in cardiomyocytes. Our results showed that overexpression of miR-215-5p could trigger myocardial autophagy through PI3K-threonine-protein kinase (AKT)-target of rapamycin (TOR) pathway. Further studies revealed that autophagic cell death was dependent on the activation of extracellular signal-regulated kinasel/2 (ERK1/2), c-Jun N-terminal kinase (JNK), p38 kinase (p38) and generation of reactive oxygen species (ROS) in overexpression of miR-215-5p in cardiomyocytes. On the contrary, miR-215-5p inhibitor can enhance cell survival capacity against autophagy by inhibiting ROS-mitogen-activated protein kinase (MAPK) pathways and activating the PI3K/AKT/TOR pathway in cardiomyocytes. Together, our findings support that miR-215-5p may modulate cell survival programs by regulating autophagy, and miR-215-5p acts as an autophagic regulator in the regulatory feedback loop that regulates cardiomyocyte survival by modulating the PI3K/AKT/TOR pathway and ROS-dependent MAPK pathways.