Abstract
Gastrodin (GAS), the principal bioactive component derived from Gastrodia elata Bl., has demonstrated efficacy in attenuating methamphetamine (MA) induced conditioned place preference (CPP) in animal models. However, the molecular mechanisms underlying its anti-addictive effects, particularly the role of miRNAs, remain insufficiently understood. A CPP model of MA dependence was established in rats. Hippocampal tissues were subjected to microarray analysis to identify differentially expressed miRNAs following GAS intervention. Key techniques including behavioral evaluation, stereotaxic injection of miR-128-3p antagomir, quantitative real time PCR, network pharmacology analysis, molecular docking, and dual-luciferase reporter assays were employed to elucidate the involvement of miR-128-3p in the anti-MA effects of GAS. GAS treatment significantly reversed MA induced CPP behaviors and mitigated the upregulation of hippocampal miR-128-3p. Inhibition of miR-128-3p via antagomir also attenuated MA induced CPP, mirroring the effect of GAS administration. Network pharmacology analysis identified 106 putative miR-128-3p targets enriched in learning, memory, and behavioral regulation pathways including PI3K Akt, MAPK, and FoxO signaling. Molecular docking revealed strong binding affinities between GAS, MA, and 6 key targets, with miR-128-3p interacting at multiple 3' UTR sites. dual-luciferase reporter assays confirmed the suppressive effect of miR-128-3p on these targets. GAS exerts its inhibitory effect on MA dependence through modulation of miR-128-3p and its downstream targets. These findings suggest that miR-128-3p represents a promising therapeutic target, and GAS may serve as a novel miRNA based intervention strategy for MA addiction.