Wang Wei, Li Chen, Sun Guangsheng, Qiu Cunxi, Fan Junyi, Jin Yuhan, Liu Kunpeng, Sun Peng
Department of Drug Prohibition and Public Security, Criminal Investigation Police University of China, Shenyang, 110035, China.
Key Laboratory of Drug Control Technology in Liaoning Province, Shenyang, 110035, China.
Sci Rep. 2024 Dec 30;14(1):31729. doi: 10.1038/s41598-024-81921-x.
Methamphetamine use disorder has emerged as a significant public health concern globally. This study endeavors to elucidate the alterations in expression changes of miRNAs in the plasma of methamphetamine use disorder and elucidate the alterations in miRNA expression in the plasma of individuals with methamphetamine use disorder and investigate the relationship between these differentially expressed miRNAs and the disorder itself, cravings for methamphetamine, and associated mental disorders. Furthermore, the study seeks to clarify the expression of downstream target molecules of specific miRNAs in the plasma of methamphetamine use disorder, assess the diagnostic utility of these miRNAs and their target molecules, explore their potential as biomarkers, and identify potential targets for the diagnosis and treatment of methamphetamine use disorder. The research subjects included 112 individuals with methamphetamine use disorder and 112 healthy controls. A questionnaire was utilized to gather baseline information on methamphetamine use disorder, assess craving for methamphetamine using a Visual Analog Scale (VAS), and evaluate anxiety (SAS) and depression (SDS) states. Employing qRT-PCR technology, we measured the expression levels of miR-9-3p and let-7b-3p in the plasma of those with the disorder. Bioinformatics tools were then used to predict downstream target molecules. Western blot analysis was conducted to quantify the levels of these target molecules in the plasma. Receiver operating characteristic (ROC) curves were utilized to evaluate the diagnostic accuracy of miRNAs and their target molecules. Additionally, we analyzed the correlation between the miRNA expression levels and the psychiatric symptoms of methamphetamine use disorder. The qRT-PCR results revealed that the expression levels of miR-9-3p and let-7b-3p were significantly elevated in the plasma of individuals with methamphetamine use disorder compared to healthy controls (P < 0.05). Furthermore, qRT-PCR and Western blot analyses demonstrated that transfection with miR-9-3p mimic led to overexpression of miR-9-3p, while transfection with let-7-3p mimic promoted overexpression of let-7-3p, concurrently inhibiting the protein levels of BDNF and GSK3B in cells (P < 0.05); ROC curve analysis indicated that the AUC for miR-9-3p was 0.782 (95% CI 0.716-0.848) and for let-7b-3p was 0.720 (95% CI 0.650-0.789). However, Spearman correlation analysis showed no significant association between the expression levels of target miRNAs and proteins and the psychiatric symptoms of methamphetamine use disorder. Notably, the combination of miR-9-3p, let-7b-3p, and BDNF exhibited high diagnostic accuracy for methamphetamine use disorder, suggesting their potential as biomarkers for its diagnosis.
甲基苯丙胺使用障碍已成为全球重大的公共卫生问题。本研究旨在阐明甲基苯丙胺使用障碍患者血浆中微小RNA（miRNA）表达变化的改变，研究这些差异表达的miRNA与该障碍本身、对甲基苯丙胺的渴望以及相关精神障碍之间的关系。此外，该研究还试图明确甲基苯丙胺使用障碍患者血浆中特定miRNA的下游靶分子的表达，评估这些miRNA及其靶分子的诊断效用，探索它们作为生物标志物的潜力，并确定甲基苯丙胺使用障碍诊断和治疗的潜在靶点。研究对象包括112名甲基苯丙胺使用障碍患者和112名健康对照者。通过问卷调查收集甲基苯丙胺使用障碍的基线信息，使用视觉模拟量表（VAS）评估对甲基苯丙胺的渴望程度，并评估焦虑（SAS）和抑郁（SDS）状态。采用qRT-PCR技术检测了患者血浆中miR-9-3p和let-7b-3p的表达水平。然后使用生物信息学工具预测下游靶分子。进行蛋白质免疫印迹分析以量化血浆中这些靶分子的水平。利用受试者工作特征（ROC）曲线评估miRNA及其靶分子的诊断准确性。此外，我们分析了miRNA表达水平与甲基苯丙胺使用障碍精神症状之间的相关性。qRT-PCR结果显示，与健康对照者相比，甲基苯丙胺使用障碍患者血浆中miR-9-3p和let-7b-3p的表达水平显著升高（P<0.05）。此外，qRT-PCR和蛋白质免疫印迹分析表明，转染miR-9-3p模拟物导致miR-9-3p过表达，而转染let-7-3p模拟物促进let-7-3p过表达，同时抑制细胞中脑源性神经营养因子（BDNF）和糖原合成酶激酶3β（GSK3B）的蛋白水平（P<0.05）；ROC曲线分析表明，miR-9-3p的曲线下面积（AUC）为0.782（95%可信区间0.716-0.848），let-7b-3p的AUC为0.720（95%可信区间0.650-0.789）。然而，Spearman相关性分析显示，靶miRNA和蛋白的表达水平与甲基苯丙胺使用障碍的精神症状之间无显著关联。值得注意的是，miR-9-3p、let-7b-3p和BDNF的联合检测对甲基苯丙胺使用障碍具有较高的诊断准确性，表明它们有潜力作为该疾病诊断的生物标志物。
