Knockdown of circHomer1 ameliorates METH-induced neuronal injury through inhibiting Bbc3 expression
Knockdown of circHomer1 ameliorates METH-induced neuronal injury through inhibiting Bbc3 expression
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敲低 circHomer1 通过抑制 Bbc3 表达改善 METH 诱导的神经元损伤
DOI：
10.1016/j.neulet.2020.135050
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发表时间：
2020-07-27
影响因子：
2.5
通讯作者：
Sun, Jinhao
中科院分区：
文献类型：
--
作者：
Li, Junwei;Sun, Qiyun;Sun, Jinhao
近年来的研究表明，环状RNA（circular RNA，circRNA）是非编码RNA（non-coding RNA，ncRNA）的重要组成部分，在脑组织中的表达量非常丰富，且呈动态变化。然而，circRNA调节甲基苯丙胺（METH）诱导的神经元损伤的确切机制仍然在很大程度上未被探索。与我们先前的研究一致，HT-22细胞中的circHomer 1的表达在METH处理后显著上调。我们通过测序用qRT-PCR产物检测其反向剪接点来证实其环结构。此外，与其线性mRNA Homer 1相比，circHomer 1对RNase R消化具有抗性。抑制circHomer 1表达确实减轻了MET诱导的神经毒性，通过流式细胞术和切割的Caspase 3蛋白水平降低了凋亡率。此外，我们推测，Bbc 3作为一个目标的circHomer 1的基础上计算算法，和敲低circHomer 1实际上减少Bbc 3的表达在mRNA和蛋白水平。此外，抑制Bbc 3可降低活性氧（ROS）水平和PI阳性细胞比率。此外，我们还利用Pearson相关系数分析了circHomer 1和Bbc 3与成熟的冰毒成瘾模型中行为的成对相关性，这表明circHomer 1和Bbc 3在成瘾行为中起着重要作用。总之，我们首次发现了一种新的circRNA，circHomer 1，我们的研究结果表明，circHomer 1通过抑制Bbc 3的表达来调节MET诱导的致死过程。
Current studies have illustrated that circular RNAs (circRNAs) are a vital part of non-coding RNA (ncRNAs) species and highly abundant and dynamically expressed in brain. However, the exact mechanisms by which circRNAs modulate methamphetamine (METH)-induced neuronal damage still remain largely unexplored. Consistent with our previous study, the expression of circHomer1 was significantly up-regulated after METH treatment in HT-22 cells. We confirmed its loop structure by detection of its back-splice junction with qRT-PCR product via sequence. Moreover, circHomer1 was resistant against RNase R digestion compared with its linear mRNA Homer1. Inhibition of circHomer1 expression indeed alleviated METH-induced neurotoxicity, with lower apoptosis rate via flow cytometry and cleaved Caspase3 protein level. Furthermore, we speculated that Bbc3 functioned as a target of circHomer1 based on computational algorithm, and knockdown of circHomer1 actually reduced Bbc3 expression at the mRNA and protein level. Besides, suppression of Bbc3 decreased the reactive oxygen species (ROS) level and radio of PI-positive cells. Furthermore, we analyzed the correlation in pairs among circHomer1, Bbc3 and behaviors in well-developed METH-addicted models using Pearson's correlation coefficient, which implied an important role of circHomer1 and Bbc3 in addictive behaviors. In all, we for the first time identified a novel circRNA, circHomer1 and our results suggested that circHomer1 regulated METH-induced lethal process by suppressing the Bbc3 expression.