Psychoactive drugs frequently contaminate aquatic environments after human consumption, raising concerns about their residues and ecological harm. This study investigates the effects of flunitrazepam (FLZ) and its metabolite 7-aminoflunitrazepam (7-FLZ), benzodiazepine-class psychoactive drugs, on brain accumulation, blood-brain barrier (BBB), and neuroinflammation of the model organism zebrafish. Molecular dynamics simulation and transcriptome sequencing were used to uncover their toxic mechanisms. Results demonstrate that both FLZ and 7-FLZ can accumulate in the brain, increasing Evans blue levels by 3.4 and 0.8 times, respectively. This increase results from abnormal expression of tight junction proteins, particularly ZO-1 and Occludin, leading to elevated BBB permeability. Furthermore, FLZ and 7-FLZ can also induce neuroinflammation, upregulating TNFα by 91% and 39%, respectively, leading to pathological changes and disrupted intracellular ion balance. Molecular dynamics simulation reveals conformational changes in ZO-1 and Occludin proteins, with FLZ exhibiting stronger binding forces and greater toxicity. Weighted gene co-expression network analysis identifies four modules correlated with BBB permeability and neuroinflammation. KEGG enrichment analysis of genes within these modules reveals pathways like protein processing in the endoplasmic reticulum, NOD-like receptor signaling pathway, and arginine and proline metabolism. This study enhances understanding of FLZ and 7-FLZ neurotoxicity and assesses environmental risks of psychoactive substances. ENVIRONMENTAL IMPLICATION: With the increasing prevalence of mental disorders and the discharge of psychoactive drugs into water, even low drug concentrations (ng/L-μg/L) can pose neurological risks. This study, utilizing molecular dynamic (MD) simulations and transcriptome sequencing, investigate the neurotoxicity and mechanisms of flunitrazepam and 7-aminoflunitrazepam. It reveals that they disrupt the blood-brain barrier in zebrafish and induce neuroinflammation primarily by inducing conformational changes in tight junction proteins. MD simulations are valuable for understanding pollutant-protein interactions. This research offers invaluable insights for the environmental risk assessment of psychoactive drugs and informs the development of strategies aimed at prevention and mitigation. Copyright © 2023 Elsevier B.V. All rights reserved.
Wenting Lin, Yingjun Qin, Yuan Ren. Flunitrazepam and its metabolites induced brain toxicity: Insights from molecular dynamics simulation and transcriptomic analysis. Journal of hazardous materials. 2024 Mar 05;465:133113
PMID: 38043427
View Full Text