Necroptosis contributes to methamphetamine-induced cytotoxicity in rat cortical neurons.
Necroptosis, a programmed necrosis, is involved in various types of neurodegenerative diseases. In this study, we investigated whether necroptosis contributed to neuronal damage in a methamphetamine injury model.


METHODS
Primary cultures of embryonic cortical neurons from Sprague-Dawley rats were subjected to different doses of methamphetamine with/without pre-treatment with a specific necroptosis inhibitor, Necrostatin-1. Necrosis was assessed by determining lactate dehydrogenase release and by Annexin V/propidium iodide double staining, while the neuronal ultra-structure was examined by electron microscopy. Tumor necrosis factor-α protein levels were determined by enzyme-linked immunosorbent assay.


RESULTS
At early stages (12h) of post-treatment with methamphetamine, significant necrosis occurred and the viability of neurons decreased in a dose- and time-dependent manner in this model of acute neuronal injury. Pretreatment with Necrostatin-1 led to significant neuronal preservation compared with the methamphetamine-treated groups. Furthermore, tumor necrosis factor-α expression increased in a dose-dependent manner following methamphetamine exposure.


CONCLUSIONS
Methamphetamine induced necrosis in rat cortical neurons in vitro, both time and dose dependently, and necroptosis may be an important newly identified mode of cortical neuronal death caused by single high-dose methamphetamine administration.
Department of Anatomy and Neurobiology, School of Basic Medical Science, Central South University, Changsha, Hunan 410013, China.
Kun
K
Xiong
Department of Forensic Science, School of Basic Medical Science, Central South University, Changsha, Hunan 410013, China.
Huidan
H
Liao
Department of Forensic Science, School of Basic Medical Science, Central South University, Changsha, Hunan 410013, China.
Lingling
L
Long
Department of Forensic Science, School of Basic Medical Science, Central South University, Changsha, Hunan 410013, China.
Yanjun
Y
Ding
Department of Anatomy and Neurobiology, School of Basic Medical Science, Central South University, Changsha, Hunan 410013, China.
Jufang
J
Huang
Department of Forensic Science, School of Basic Medical Science, Central South University, Changsha, Hunan 410013, China. Electronic address: wills212156@gmail.com.
Jie
J
Yan
D000818:Animals; D016923:Cell Death / Q000187:drug effects; D005260:Female; D007093:Imidazoles / Q000378:metabolism; D007211:Indoles / Q000378:metabolism; D008694:Methamphetamine / Q000633:toxicity*; D008854:Microscopy, Electron; D009474:Neurons / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology / Q000648:ultrastructure; D051381:Rats; D017207:Rats, Sprague-Dawley; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism
D007093:Imidazoles; D007211:Indoles; D014409:Tumor Necrosis Factor-alpha; C507699:necrostatin-1; D008694:Methamphetamine
Embryonic cortical neurons; Methamphetamine; Necroptosis; Necrosis; Neurotoxicity