Neural Regeneration Research ›› 2020, Vol. 15 ›› Issue (5): 865-874.doi: 10.4103/1673-5374.268902

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RIP3/MLKL-mediated neuronal necroptosis induced by methamphetamine at 39°C

Li-Min Guo1, Zhen Wang1, 2, Shi-Ping Li3, Mi Wang1, Wei-Tao Yan1, Feng-Xia Liu4, Chu-Dong Wang5, Xu-Dong Zhang6, Dan Chen1, Jie Yan5, Kun Xiong1#br#   

  1. 1 Department of Neurobiology and Human Anatomy, School of Basic Medical Science, Central South University, Changsha, Hunan Province, China
    2 Department of Basic Medicine, Wuxi School of Medicine, Jiangnan University, Wuxi, Jiangsu Province, China
    3 Department of Neurology, People’s Hospital of Lianhua, Pingxiang, Jiangxi Province, China
    4 Department of Human Anatomy, School of Basic Medical Science, Xinjiang Medical University, Urumqi, Xinjiang Uygur Autonomous Region, China
    5 Department of Forensic Science, School of Basic Medical Science, Central South University, Changsha, Hunan Province, China
    6 Narcotics Division, Municipal Security Bureau, Changsha, Hunan Province, China
  • Online:2020-05-15 Published:2020-06-01
  • Contact: Kun Xiong, MD,xiongkun2001@163.com; Jie Yan, wills212156@csu.edu.cn.
  • Supported by:
    This study was funded by the National Natural Science Foundation of China, No. 81971891 (to KX), 81571939 (to KX), 81772134 (to KX), 81772024 (to JY), and 81860781 (to FXL); the Key Research and Development Program of Hunan Province of China, No. 2018SK2091 (to KX); the Natural Science Foundation of Hunan Province of China, No. 2017JJ2339 (to JY); the Wu Jie-Ping Medical Foundation of the Minister of Health of China, No. 320.6750.14118 (to KX).

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Abstract: Methamphetamine is one of the most prevalent drugs abused in the world. Methamphetamine abusers usually present with hyperpyrexia (39°C), hallucination and other psychiatric symptoms. However, the detailed mechanism underlying its neurotoxic action remains elusive. This study investigated the effects of methamphetamine + 39°C on primary cortical neurons from the cortex of embryonic Sprague-Dawley rats. Primary cortex neurons were exposed to 1 mM methamphetamine + 39°C. Propidium iodide staining and lactate dehydrogenase release detection showed that methamphetamine + 39°C triggered obvious necrosis-like death in cultured primary cortical neurons, which could be partially inhibited by receptor-interacting protein-1 (RIP1) inhibitor Necrostatin-1 partially. Western blot assay results showed that there were increases in the expressions of receptor-interacting protein-3 (RIP3) and mixed lineage kinase domain-like protein (MLKL) in the primary cortical neurons treated with 1 mM methamphetamine + 39°C for 3 hours. After pre-treatment with RIP3 inhibitor GSK’872, propidium iodide staining and lactate dehydrogenase release detection showed that neuronal necrosis rate was significantly decreased; RIP3 and MLKL protein expression significantly decreased. Immunohistochemistry staining results also showed that the expressions of RIP3 and MLKL were up-regulated in brain specimens from humans who had died of methamphetamine abuse. Taken together, the above results suggest that methamphetamine + 39°C can induce RIP3/MLKL regulated necroptosis, thereby resulting in neurotoxicity. The study protocol was approved by the Medical Ethics Committee of the Third Xiangya Hospital of Central South University, China (approval numbers: 2017-S026 and 2017-S033) on March 7, 2017.

Key words: GSK’872, human brain tissue, hyperpyrexia, methamphetamine, mixed lineage kinase domain-like protein, necrostatin-1, necroptosis, nerve regeneration, neural regeneration, rat cortical neurons, receptor-interacting protein-3, synergistic effect