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.

当前的研究表明，环状RNA（circRNA）是非编码RNA（ncRNA）物种的重要组成部分，在大脑中高度丰富且动态表达。但是，circRNA调节甲基苯丙胺（METH）诱导的神经元损伤的确切机制仍在很大程度上尚待探索。与我们以前的研究一致，在HT-22细胞中，经METH处理后，circHomer1的表达明显上调。我们通过序列检测其与qRT-PCR产物的反向剪接连接来确认其环结构。此外，circHomer1与其线性mRNA Homer1相比，对RNase R消化具有抵抗力。抑制circHomer1表达确实减轻了METH诱导的神经毒性，通过流式细胞仪检测了较低的细胞凋亡率，并裂解了Caspase3蛋白水平。此外，我们基于计算算法推测Bbc3作为circHomer1的靶标，而敲除circHomer1实际上降低了mRNA和蛋白质水平上Bbc3的表达。此外，Bbc3的抑制降低了PI阳性细胞的活性氧（ROS）水平和放射。此外，我们使用皮尔逊相关系数分析了发达的METH上瘾模型中circHomer1，Bbc3与行为之间的成对相关性，这暗示了circHomer1，Bbc3在成瘾行为中的重要作用。总而言之，我们首次鉴定出一种新型的circRNA，即circHomer1，我们的研究结果表明，circHomer1通过抑制Bbc3表达来调节METH诱导的致死过程。circHomer1的敲低实际上降低了mRNA和蛋白质水平的Bbc3表达。此外，Bbc3的抑制降低了PI阳性细胞的活性氧（ROS）水平和放射。此外，我们使用皮尔逊相关系数分析了发达的METH上瘾模型中circHomer1，Bbc3与行为之间的成对相关性，这暗示了circHomer1，Bbc3在成瘾行为中的重要作用。总而言之，我们首次鉴定出一种新型的circRNA，即circHomer1，我们的研究结果表明circHomer1通过抑制Bbc3表达来调节METH诱导的致死过程。circHomer1的敲低实际上降低了mRNA和蛋白质水平的Bbc3表达。此外，Bbc3的抑制降低了PI阳性细胞的活性氧（ROS）水平和放射。此外，我们使用皮尔逊相关系数分析了发达的METH上瘾模型中circHomer1，Bbc3与行为之间的成对相关性，这暗示了circHomer1，Bbc3在成瘾行为中的重要作用。总而言之，我们首次鉴定出一种新型的circRNA，即circHomer1，我们的研究结果表明circHomer1通过抑制Bbc3表达来调节METH诱导的致死过程。使用皮尔逊相关系数在发达的METH上瘾模型中，Bbc3和行为，这暗示circHomer1和Bbc3在成瘾行为中起重要作用。总而言之，我们首次鉴定出一种新型的circRNA，即circHomer1，我们的研究结果表明circHomer1通过抑制Bbc3表达来调节METH诱导的致死过程。使用皮尔逊相关系数在发达的METH上瘾模型中，Bbc3和行为，这暗示circHomer1和Bbc3在成瘾行为中起重要作用。总而言之，我们首次鉴定出一种新型的circRNA，即circHomer1，我们的研究结果表明circHomer1通过抑制Bbc3表达来调节METH诱导的致死过程。