College of Forensic Medicine, Xi'an Jiaotong University Health Science Center, Xi'an 710061, China.
The Key Laboratory of Health Ministry for Forensic Science, Xi'an Jiaotong University, Xi'an 710061, China.
Int J Mol Sci. 2024 Jan 11;25(2):942. doi: 10.3390/ijms25020942.
Methamphetamine (METH) abuse inflicts both physical and psychological harm. While our previous research has established the regulatory role of miR-29c-3p in behavior sensitization, the underlying mechanisms and target genes remain incompletely understood. In this study, we employed the isobaric tags for relative and absolute quantitation (iTRAQ) technique in conjunction with Ingenuity pathway analysis (IPA) to probe the putative molecular mechanisms of METH sensitization through miR-29c-3p inhibition. Through a microinjection of AAV-anti-miR-29c-3p into the nucleus accumbens (NAc) of mice, we observed the attenuation of METH-induced locomotor effects. Subsequent iTRAQ analysis identified 70 differentially expressed proteins (DEPs), with 22 up-regulated potential target proteins identified through miR-29c-3p target gene prediction and IPA analysis. Our focus extended to the number of neuronal branches, the excitatory synapse count, and locomotion-related pathways. Notably, GPR37, NPC1, and IREB2 emerged as potential target molecules for miR-29c-3p regulation, suggesting their involvement in the modulation of METH sensitization. Quantitative PCR confirmed the METH-induced aberrant expression of , , and in the NAc of mice. Specifically, the over-expression of miR-29c-3p led to a significant reduction in the mRNA level of , while the inhibition of miR-29c-3p resulted in a significant increase in the mRNA level of , consistent with the regulatory principle of miRNAs modulating target gene expression. This suggests that miR-29c-3p potentially influences METH sensitization through its regulation of neuroplasticity. Our research indicates that miR-29c-3p plays a crucial role in regulating METH-induced sensitization, and it identified the potential molecular of miR-29c-3p in regulating METH-induced sensitization.
甲基苯丙胺（METH）滥用会造成身体和心理伤害。虽然我们之前的研究已经确定了 miR-29c-3p 在行为敏化中的调节作用，但潜在的机制和靶基因仍不完全清楚。在这项研究中，我们采用相对和绝对定量同位素标记（iTRAQ）技术结合 Ingenuity 通路分析（IPA），通过抑制 miR-29c-3p 来探究 METH 敏化的潜在分子机制。通过将 AAV-anti-miR-29c-3p 微注射到小鼠伏隔核（NAc）中，我们观察到 METH 诱导的运动效应减弱。随后的 iTRAQ 分析鉴定出 70 个差异表达蛋白（DEPs），通过 miR-29c-3p 靶基因预测和 IPA 分析鉴定出 22 个上调的潜在靶蛋白。我们的研究重点扩展到神经元分支数量、兴奋性突触计数和与运动相关的通路。值得注意的是，GPR37、NPC1 和 IREB2 成为 miR-29c-3p 调节的潜在靶分子，表明它们参与了 METH 敏化的调节。定量 PCR 证实了 miR-29c-3p 在小鼠 NAc 中诱导的 、 和 异常表达。具体而言，miR-29c-3p 的过表达导致 的 mRNA 水平显著降低，而 miR-29c-3p 的抑制导致 的 mRNA 水平显著增加，这与 miRNA 调节靶基因表达的调节原理一致。这表明 miR-29c-3p 通过调节神经可塑性潜在地影响 METH 敏化。我们的研究表明，miR-29c-3p 在调节 METH 诱导的敏化中起着至关重要的作用，并确定了 miR-29c-3p 在调节 METH 诱导的敏化中的潜在分子。
