Zhou Yun, Hong Qingxiao, Xu Wenjin, Chen Weisheng, Xie Xiaohu, Zhuang Dingding, Lai Miaojun, Fu Dan, Xu Zemin, Wang Majie, Zhou Wenhua, Liu Huifen
School of Medicine, Ningbo University, Laboratory of Behavioral Neuroscience, Ningbo Kangning Hospital, Ningbo, Zhejiang, China.
Key Laboratory of Addiction Research of Zhejiang Province, Ningbo, Zhejiang, China.
Front Genet. 2023 Feb 2;14:1088498. doi: 10.3389/fgene.2023.1088498. eCollection 2023.
Transfer RNA-derived small RNAs (tsRNAs) are a novel class of short, non-coding RNAs that are closely associated with the pathogenesis of various diseases. Accumulating evidence has demonstrated their critical functional roles as regulatory factors in gene expression regulation, protein translation regulation, regulation of various cellular activities, immune mediation, and response to stress. However, the underlying mechanisms by which tRFs & tiRNAs affect methamphetamine-induced pathophysiological processes are largely unknown. In this study, we used a combination of small RNA sequencing, quantitative reverse transcription-polymerase chain reaction (qRT‒PCR), bioinformatics, and luciferase reporter assays to screen the expression profiles and identify the functional roles of tRFs and tiRNAs in the nucleus accumbens (NAc) of methamphetamine self-administration rat models. A total of 461 tRFs & tiRNAs were identified in the NAc of rats after 14 days of methamphetamine self-administration training. Of those, 132 tRFs & tiRNAs were significantly differentially expressed: 59 were significantly upregulated, whereas 73 were significantly downregulated in the rats with methamphetamine self-administration. Decreased expression levels of tiRNA-1-34-Lys-CTT-1 and tRF-1-32-Gly-GCC-2-M2, as well as increased expression levels of tRF-1-16-Ala-TGC-4 in the METH group compared with the saline control were validated by using RT‒PCR. Then, bioinformatic analysis was performed to analyse the possible biological functions of tRFs & tiRNAs in methamphetamine-induced pathogenesis. Furthermore, tRF-1-32-Gly-GCC-2-M2 was identified to target BDNF using the luciferase reporter assay. An altered tsRNA expression pattern was proven, and tRF-1-32-Gly-GCC-2-M2 was shown to be involved in methamphetamine-induced pathophysiologic processes by targeting BDNF. The current study provides new insights for future investigations to explore the mechanisms and therapeutic methods for methamphetamine addiction.
转运RNA衍生的小RNA（tsRNAs）是一类新型的短链非编码RNA，与多种疾病的发病机制密切相关。越来越多的证据表明，它们作为调节因子在基因表达调控、蛋白质翻译调控、各种细胞活动调节、免疫介导和应激反应中发挥着关键的功能作用。然而，tRFs和tiRNAs影响甲基苯丙胺诱导的病理生理过程的潜在机制在很大程度上尚不清楚。在本研究中，我们结合小RNA测序、定量逆转录-聚合酶链反应（qRT-PCR）、生物信息学和荧光素酶报告基因检测，以筛选甲基苯丙胺自我给药大鼠模型伏隔核（NAc）中tRFs和tiRNAs的表达谱并确定其功能作用。在甲基苯丙胺自我给药训练14天后，在大鼠的NAc中共鉴定出461种tRFs和tiRNAs。其中，132种tRFs和tiRNAs有显著差异表达：59种显著上调，而73种在甲基苯丙胺自我给药的大鼠中显著下调。通过RT-PCR验证了与生理盐水对照组相比，甲基苯丙胺组中tiRNA-1-34-Lys-CTT-1和tRF-1-32-Gly-GCC-2-M2的表达水平降低，以及tRF-1-16-Ala-TGC-4的表达水平升高。然后，进行生物信息学分析以分析tRFs和tiRNAs在甲基苯丙胺诱导的发病机制中的可能生物学功能。此外，通过荧光素酶报告基因检测确定tRF-1-32-Gly-GCC-2-M2靶向脑源性神经营养因子（BDNF）。证实了tsRNA表达模式的改变，并且tRF-1-32-Gly-GCC-2-M2通过靶向BDNF参与甲基苯丙胺诱导的病理生理过程。本研究为未来探索甲基苯丙胺成瘾的机制和治疗方法提供了新的见解。
