Liu Tian-Cong, Li Hong-Xi, Wan Yu-Xiao, Shi Guang, Zhao Yun-Peng, Liu Yi-Fei, Fan Xin-Yu
Department of Otolaryngology, Shengjing Hospital of China Medical University, Shenyang, China.
Department of Pain Management, Shengjing Hospital of China Medical University, Shenyang, China.
CNS Neurosci Ther. 2024 Jul;30(7):e14830. doi: 10.1111/cns.14830.
N6-methyladenosine (m6A) methylation is a vital epigenetic mechanism associated with drug addiction. However, the relationship between m6A modification and oxycodone rewarding is less well explored. Based on an open field test, the present study evaluated oxycodone rewarding using chromatin immunoprecipitation PCR, immunofluorescence, and RNA sequencing. A marked increase in METTL14 protein and a decrease in PP1α protein due to oxycodone abundance in the striatal neurons were observed in a dose- and time-dependent manner. Oxycodone markedly increased LSD1 expression, and decreased H3K4me1 expression in the striatum. In the open field test, intra-striatal injection of METTL14 siRNA, HOTAIR siRNA, or LSD1 shRNA blocked oxycodone-induced increase in locomotor activity. The downregulation of PP1α was also inhibited after treatment with METTL14/HOTAIR siRNA and LSD1 shRNA. Enhanced binding of LSD1 with CoRest and of CoRest with the PP1α gene induced by oxycodone was also reversed by LSD1 shRNA. In addition, H3K4me1 demethylation was also blocked by the treatment. In summary, the investigation confirmed that METTL14-mediated upregulation of HOTAIR resulted in the repression of PP1α, which in turn facilitated the recruitment of LSD1, thus catalyzing H3K4me1 demethylation and promoting oxycodone addiction.
N6-甲基腺苷(m6A)甲基化是一种与药物成瘾相关的重要表观遗传机制。然而,m6A修饰与羟考酮奖赏效应之间的关系尚未得到充分研究。基于旷场试验,本研究采用染色质免疫沉淀PCR、免疫荧光和RNA测序技术评估羟考酮的奖赏效应。结果发现,纹状体神经元中,随着羟考酮含量的增加,METTL14蛋白显著增加,PP1α蛋白显著减少,且呈剂量和时间依赖性。羟考酮显著增加纹状体中LSD1的表达,并降低H3K4me1的表达。在旷场试验中,纹状体内注射METTL14 siRNA、HOTAIR siRNA或LSD1 shRNA可阻断羟考酮诱导的运动活性增加。METTL14/HOTAIR siRNA和LSD1 shRNA处理后,PP1α的下调也受到抑制。LSD1 shRNA还可逆转羟考酮诱导的LSD1与CoRest以及CoRest与PP1α基因结合增强的现象。此外,该处理还可阻断H3K4me1的去甲基化。综上所述,本研究证实METTL14介导的HOTAIR上调导致PP1α受到抑制,进而促进LSD1的募集,从而催化H3K4me1去甲基化并促进羟考酮成瘾。