Li Hancheng, Peng Yongen, Wu Yangkai, Chen Yiling, Li Jieyu, He Yunbing, Wang Hongwu, Luo Chaohua, Mo Zhixian
Department of Pharmaceutical Engineering, School of Food and Pharmaceutical Engineering, Zhaoqing University, Zhaoqing, China.
Department of Pharmacology of Chinese Medicine, School of Traditional Chinese Medicine, Southern Medical University, Guangzhou, China.
Front Pharmacol. 2025 May 14;16:1541442. doi: 10.3389/fphar.2025.1541442. eCollection 2025.
Methamphetamine (MA) is one of the most harmful synthetic drugs, yet the mechanisms underlying its addiction and relapse remain incompletely understood. This study investigates how cardiomyocyte-derived exosomes carrying miRNAs facilitate heart-brain crosstalk and contribute to MA dependence.
A conditioned place preference (CPP) model of MA dependence was established in rats. High-throughput sequencing were employed to identify candidate miRNAs in cardiac exosomes and brain tissues. Behavioral assessments, real-time PCR, nanoparticle tracking analysis, imaging, uptake assays, network pharmacology, and dual-luciferase reporter assays were used to explore the role of cardiomyocyte-derived exosomes in MA dependence.
MA induced significant CPP in rats. miR-181a-5p was markedly upregulated in cardiac exosomes and brain tissue, with higher levels observed in cardiac exosomes. biodistribution showed that cardiomyocyte-derived exosomes cross the blood-brain barrier and accumulate in the brain. uptake assays demonstrated that SH-SY5Y cells internalized these exosomes, leading to increased miR-181a-5p expression. Tail vein injections of miR-181a-5p-enriched exosomes enhanced MA CPP behavior in rats. Network pharmacology revealed 108 potential targets of miR-181a-5p, enriched in processes such as steroid biosynthesis, amide metabolism, and apoptosis, involving pathways related to the endoplasmic reticulum, MAPK signaling, and amyotrophic lateral sclerosis. Molecular docking identified stable interactions between MA and 12 targets, including HSP90B1, TNF, and MAP2K1, with miR-181a-5p binding to the 3'-UTR regions of these targets. Dual-luciferase assays confirmed the negative regulation of six targets by miR-181a-5p.
This study reveals that cardiomyocyte-derived exosomes transport miR-181a-5p, facilitating heart-brain crosstalk and exacerbating MA CPP behavior in rats. These effects are mediated through the regulation of key brain targets, including HSP90B1, TNF, and MAP2K1, providing new insights into the molecular mechanisms of MA addiction and potential therapeutic targets.
甲基苯丙胺（MA）是最有害的合成毒品之一，但其成瘾和复发的潜在机制仍未完全了解。本研究调查携带微小RNA（miRNA）的心肌细胞衍生外泌体如何促进心脏与大脑的相互作用并导致MA依赖。
在大鼠中建立MA依赖的条件性位置偏爱（CPP）模型。采用高通量测序来鉴定心脏外泌体和脑组织中的候选miRNA。使用行为评估、实时聚合酶链反应（PCR）、纳米颗粒跟踪分析、成像、摄取试验、网络药理学和双荧光素酶报告基因试验来探究心肌细胞衍生外泌体在MA依赖中的作用。
MA在大鼠中诱导出显著的CPP。miR-181a-5p在心脏外泌体和脑组织中显著上调，在心脏外泌体中观察到更高水平。生物分布显示心肌细胞衍生的外泌体穿过血脑屏障并在脑中积累。摄取试验表明SH-SY5Y细胞摄取这些外泌体，导致miR-181a-5p表达增加。尾静脉注射富含miR-181a-5p的外泌体增强了大鼠的MA CPP行为。网络药理学揭示了miR-181a-5p的108个潜在靶点，富集于类固醇生物合成、酰胺代谢和凋亡等过程，涉及与内质网、丝裂原活化蛋白激酶（MAPK）信号传导和肌萎缩侧索硬化相关的途径。分子对接确定了MA与12个靶点之间的稳定相互作用，包括热休克蛋白90β1（HSP90B1）、肿瘤坏死因子（TNF）和丝裂原活化蛋白激酶1（MAP2K1），miR-181a-5p与这些靶点的3'-非翻译区（3'-UTR）结合。双荧光素酶试验证实miR-181a-5p对六个靶点具有负调控作用。
本研究表明心肌细胞衍生的外泌体运输miR-181a-5p，促进心脏与大脑的相互作用并加剧大鼠的MA CPP行为。这些作用是通过调节关键脑靶点介导的，包括HSP90B1、TNF和MAP2K1，为MA成瘾的分子机制和潜在治疗靶点提供了新的见解。
