2. Academic Validation
  3. Chlorpyrifos-oxon results in autophagic flux dysfunction contributing to neuronal apoptosis via a ROS/AMPK/CHOP activation pathway

Chlorpyrifos-oxon results in autophagic flux dysfunction contributing to neuronal apoptosis via a ROS/AMPK/CHOP activation pathway

  • Chem Biol Interact. 2025 May 1:412:111452. doi: 10.1016/j.cbi.2025.111452.
Jui-Ming Liu 1,  Kuan-I Lee 2,  Chin-Chuan Su 3,  Kai-Min Fang 4,  Shing-Hwa Liu 5,  Shih-Chang Fu 6,  Chun-Ying Kuo 7,  Kai-Chih Chang 8,  Jun-An Ke 9,  Ya-Wen Chen 10,  Ching-Yao Yang 11,  Chun-Fa Huang 12
Affiliations

Affiliations

  • 1 Division of Urology, Department of Surgery, Taoyuan General Hospital, Ministry of Health and Welfare, Taoyuan, 330, Taiwan; Department of Obstetrics and Gynecology, Tri-Service General Hospital, National Defense Medical Center, Taipei, 114 Taiwan.
  • 2 Department of Emergency, Taichung Tzu Chi Hospital, Buddhist Tzu Chi Medical Foundation, Taichung, 427, Taiwan.
  • 3 Department of Otorhinolaryngology, Head and Neck Surgery, Changhua Christian Hospital, Changhua City, 50006, Taiwan; Department of Post-Baccalaureate Medicine, College of Medicine, National Chung Hsing University, Taichung, 402202, Taiwan.
  • 4 Department of Otolaryngology, Far Eastern Memorial Hospital, New Taipei City, 220, Taiwan.
  • 5 Institute of Toxicology, College of Medicine, National Taiwan University, Taipei, 100, Taiwan.
  • 6 Division of Urology, Department of Surgery, Taoyuan General Hospital, Ministry of Health and Welfare, Taoyuan, 330, Taiwan.
  • 7 Department of Otorhinolaryngology, Head and Neck Surgery, Changhua Christian Hospital, Changhua City, 50006, Taiwan.
  • 8 Center for Digestive Medicine, Department of Internal Medicine, China Medical University Hospital, Taichung, 404, Taiwan.
  • 9 Department of Medical Education, Changhua Christian HospitalChanghua City, 500, Taiwan.
  • 10 Department of Physiology, School of Medicine, College of Medicine, China Medical University, Taichung, 404, Taiwan.
  • 11 Department of Surgery, National Taiwan University Hospital, And Department of Surgery, College of Medicine, National Taiwan University, Taipei, 100, Taiwan. Electronic address: cyang@ntuh.gov.tw.
  • 12 School of Chinese Medicine, College of Chinese Medicine, China Medical University, Taichung, 404, Taiwan; Department of Nursing, College of Medical and Health Science, Asia University, Taichung, 413, Taiwan. Electronic address: cfhuang@mail.cmu.edu.tw.
PMID: 40049439 DOI: 10.1016/j.cbi.2025.111452
Abstract

Chlorpyrifos (CPF) is a widely used organophosphate (OP) pesticide in agriculture and sanitation, known to elicit neurotoxic effects. Chlorpyrifos-oxon (CPO), a metabolite of CPF, is the primary neurotoxic agent, yet its mechanisms are less understood. In this study, we investigated the effects and underlying mechanisms of CPO-induced neurotoxicity. CPO exposure significantly induced cytotoxicity in Neuro-2a cells, alongside the activation of Apoptosis, as evidenced by an increase in the apoptotic cell population, Caspase-3 activity, and cleavage of caspaspe-3, -7, and PARP proteins. Furthermore, defective Autophagy was observed in CPO-treated Neuro-2a cells, indicated by increased expression of Beclin-1, Atg5, LC3-II, and p62 proteins. 3-MA, an Autophagy Inhibitor, suppressed CPO-activated LC3-II and apoptotic marker proteins expression, but not p62. In contrast, chloroquine and bafilomycin A1, autophagic flux inhibitors, potentiated the CPO-induced elevation of LC3-II, p62, and cleaved Caspase-3 and -7 protein levels. CPO exposure also upregulated CHOP protein expression. Transfection with CHOP-specific siRNA markedly reduced CHOP protein expression, autophagic flux dysfunction, and Apoptosis. Additionally, CPO exposure significantly increased AMPKα phosphorylation and Reactive Oxygen Species (ROS) generation. Antioxidant N-acetylcysteine (NAC), but not the AMPK Inhibitor Compound C, effectively attenuated the CPO-induced ROS generation in neuronal cells, which was accompanied by the prevention of AMPKα activation, downstream CHOP expression, autophagic flux dysfunction, and Apoptosis. Collectively, these findings suggest that CPO-induced neurotoxicity arises from autophagic flux dysfunction, contributing to Apoptosis via the ROS-activated AMPK pathway, which regulates CHOP expression, ultimately leading to neuronal cell death. Targeting the ROS/AMPK/CHOP axis may offer a promising intervention to against CPO-induced neurotoxicity.

Keywords

AMPKα; Apoptosis; Autophagic flux; CHOP; Chlorpyrifos-oxon (CPO); Neurotoxicity; Reactive oxygen species.

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