Sakai Yuji, Hattori Junta, Morikawa Yoshifumi, Matsumura Toshihiro, Jimbo Shunsuke, Suenami Koichi, Takayama Tomohiro, Nagai Atsushi, Michiue Tomomi, Ikari Akira, Matsunaga Toshiyuki
Forensic Science Laboratory, Gifu Prefectural Police Headquarters, Gifu, 500-8501, Japan.
Laboratory of Bioinformatics, Gifu Pharmaceutical University, Gifu, 502-8585, Japan.
Forensic Toxicol. 2025 Jan;43(1):142-154. doi: 10.1007/s11419-024-00708-x. Epub 2024 Dec 9.
Pyrrolidinophenone derivatives (PPs) are amphetamine-like designer drugs containing a pyrrolidine ring, and their adverse effects resemble those of methamphetamine (METH). Microglial activation has been recently suggested as a key event in eliciting the adverse effects against dysfunction of the central nervous system. The aim of this study is to clarify the mechanisms of microglial activation induced by PPs.
We employed the human microglial cell line HMC3 to assess microglial activation induced by PPs and evaluated the capacities for proliferation and interleukin-6 (IL-6) production that are characteristic features of the activation events.
The WST-1 assay indicated that viability of HMC3 cells was increased by treatment with sublethal concentrations (5-20 µM) of α-pyrrolidinooctanophenone (α-POP), a highly lipophilic PP, whereas it was decreased by treatment with concentrations above 40 µM. Treatment with sublethal α-POP concentrations up-regulated the expression and secretion of IL-6. Additionally, α-POP-induced increase in cell viability was restored by pretreating with N-acetyl-L-cysteine, a reactive oxygen species (ROS) scavenger, and stattic, an inhibitor of signal transducer and activator of transcription 3 (STAT3), respectively, suggesting that activation of the ROS/STAT3 pathway is involved in the α-POP-induced activation of HMC3 cells. The increases in cell viability were also observed in HMC3 cells treated with other α-POP derivatives and METH.
These results suggest that enhanced productions of ROS and IL-6 are also involved in microglial activation by drug treatment and that HMC3 cell-based system is available to evaluate accurately the microglial activation induced by abused drugs.
吡咯烷苯酮衍生物（PPs）是一类含有吡咯烷环的苯丙胺类新型毒品，其不良反应与甲基苯丙胺（METH）相似。近期研究表明，小胶质细胞激活是引发中枢神经系统功能障碍不良反应的关键事件。本研究旨在阐明PPs诱导小胶质细胞激活的机制。
我们使用人小胶质细胞系HMC3来评估PPs诱导的小胶质细胞激活，并评估了增殖能力和白细胞介素-6（IL-6）产生情况，这些都是激活事件的特征。
WST-1检测表明，用亚致死浓度（5-20µM）的α-吡咯烷辛酰苯（α-POP，一种高度亲脂性的PP）处理可提高HMC3细胞的活力，而浓度高于40µM时则会降低细胞活力。用亚致死浓度的α-POP处理可上调IL-6的表达和分泌。此外，分别用活性氧（ROS）清除剂N-乙酰-L-半胱氨酸和信号转导子及转录激活子3（STAT3）抑制剂stattic预处理可恢复α-POP诱导的细胞活力增加，这表明ROS/STAT3通路的激活参与了α-POP诱导的HMC3细胞激活。在用其他α-POP衍生物和METH处理的HMC3细胞中也观察到了细胞活力的增加。
这些结果表明，ROS和IL-6的产生增加也参与了药物治疗诱导的小胶质细胞激活，并且基于HMC3细胞的系统可用于准确评估滥用药物诱导的小胶质细胞激活。
