Department of Psychiatry, University of Ulsan College of Medicine, Asan Medical Center, Seoul, Korea.
Neuroscience. 2012 May 3;209:64-73. doi: 10.1016/j.neuroscience.2012.02.035. Epub 2012 Feb 24.
Early intervention and maintenance treatment for schizophrenia patients may prolong the duration of exposure to antipsychotic agents; however, there have been few studies on the neurotoxicity of these agents. Here, we investigated the effects of antipsychotics on cell viability and autophagy in rat primary neurons. Cultured cortical neurons obtained from rat embryos were treated with various concentrations of haloperidol and clozapine, and the neuronal toxicity was assessed by measuring lactate dehydrogenase (LDH) activity and 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay. Autophagosomes were quantitated by measuring the level of microtubule-associated protein 1A/1B-light chain 3 (LC3-II) by Western blot and immunofluorescence staining. Autophagic flux was assayed using bafilomycin A1 and GFP-mCherry-LC3 transfection. Haloperidol and clozapine decreased the viability of neurons in vitro in a concentration- and time-dependent manner. We also observed increased accumulation of autophagosomes after antipsychotic treatment. Using bafilomycin A1 and GFP-mCherry-LC3 transfection, we discovered that haloperidol and clozapine inhibited autophagosome turnover resulting in a dysfunctional autophagic process, including impaired lysosomal fusion. Together, these results suggest that haloperidol and clozapine negatively affect neuronal viability, possibly by blocking autophagolysosome formation.
抗精神病药物早期干预和维持治疗可能会延长抗精神病药物暴露时间；然而，关于这些药物的神经毒性的研究很少。在这里，我们研究了抗精神病药对大鼠原代神经元细胞活力和自噬的影响。用不同浓度的氟哌啶醇和氯氮平处理从大鼠胚胎中获得的培养皮质神经元，并通过测量乳酸脱氢酶（LDH）活性和 3-(4,5-二甲基噻唑-2-基）-5-(3-羧甲基氧苯基）-2-(4-磺基苯基）-2H-四唑（MTS）测定来评估神经元毒性。通过 Western blot 和免疫荧光染色测量微管相关蛋白 1A/1B-轻链 3（LC3-II）的水平来定量自噬体。使用巴弗洛霉素 A1 和 GFP-mCherry-LC3 转染测定自噬通量。氟哌啶醇和氯氮平以浓度和时间依赖的方式体外降低神经元的活力。我们还观察到抗精神病药物治疗后自噬体的积累增加。使用巴弗洛霉素 A1 和 GFP-mCherry-LC3 转染，我们发现氟哌啶醇和氯氮平抑制自噬体的周转，导致自噬过程功能障碍，包括溶酶体融合受损。总之，这些结果表明氟哌啶醇和氯氮平可能通过阻断自噬溶酶体形成来负性影响神经元活力。
