Nrf2缺失影响甲基苯丙胺致前额叶皮质ASIC表达及神经元损伤
Effects of Nrf2 deficiency on ASIC expressions and neuronal injury in the methamphetamine-exposed prefrontal cortex
摘要
Abstract
OBJECTIVE To investigate the effect of nuclear factor erythroid 2-related factor 2(Nrf2)deficiency on methamphetamine(METH)-induced changes in expressions of acid-sensing ion channel 1α(ASIC1α)and ASIC2 and on neuronal injury in the prefrontal cortex.METHODS ① Immunofluores-cence was used to detect Nrf2 and ASIC1α expressions in primary cortical neurons after 24 hours of treatment with METH at 1 mmol·L-1.An Nrf2 knockdown model was established in the mouse hippo-campal neuronal cell line HT22 by using short hairpin RNA targeting Nrf2(sh-Nrf2).After 24 hours of treatment with METH at 1 mmol·L-1,ASIC1α and ASIC2 protein expressions were detected by Western blotting.② The genotypes of mice were identified using a genotyping kit before male C57BL/6J wild-type(WT)and Nrf2-knockout(Nrf2-KO)mice were randomly assigned to a control group and a METH group.A repeated METH exposure model was established using an escalating-dose regimen in which METH and physiological saline were administered on alternating days.Scores of conditioned place preference(CPP)and rates of spontaneous alternation were determined in mice using the CPP test and Y-maze test,respectively.Double immunofluorescence staining was used to examine the co-expres-sion of ASIC1α+/vesicular glutamate transporter 1(VGLUT-1)+and ASIC2+/VGLUT-1+in VGLUT-1+neurons.HE staining and Nissl staining were performed to assess morphological injury to neurons.In WT mice,Western blotting was used to determine the protein expressions of total Nrf2(T-Nrf2),nuclear Nrf2(N-Nrf2),ASIC1α and ASIC2 in the prefrontal cortex while Western blotting and immunohistochemistry were adopted to determine ASIC1α and ASIC2 expressions in the prefrontal cortex of WT and Nrf2-KO mice.RESULTS ① After 24 hours of treatment with METH,Nrf2 and ASIC1α expressions were signifi-cantly increased in primary cortical neurons.Compared with the sh-NC-METH group,ASIC1α and ASIC2 protein expressions were significantly decreased in HT22 cells in the sh-Nrf2-METH group.② Behavioral results showed that CPP scores were increased in the post-test after METH exposure,accompanied by a decrease in spontaneous alternation in the Y-maze test.Compared with the WT-METH group,spontaneous alternation kept decreasing in the Nrf2-KO-METH group.Compared with the WT-control group,the proportions of ASIC1α+/VGLUT-1+and ASIC2+/VGLUT-1+neurons were significantly increased in the WT-METH group.HE staining and Nissl staining suggested that METH expo-sure induced neuronal morphological injury in the prefrontal cortex,characterized by neuronal shrinkage,nuclear hyperchromasia and an increased percentage of shrunken neurons.Neuronal injury was aggra-vated in the Nrf2-KO-METH group compared with the WT-METH group.Compared with the WT-control group,protein expressions of T-Nrf2,N-Nrf2,ASIC1α and ASIC2 in the prefrontal cortex were signif-icantly increased in the WT-METH group.Compared with the WT-METH group,ASIC1α and ASIC2 protein expressions and percentages of positive staining areas in the prefrontal cortex were signif-icantly decreased in the Nrf2-KO-METH group.CONCLUSION METH can activate Nrf2,upregulate ASIC1α and ASIC2,and cause neuronal injury in the prefrontal cortex.Nrf2 deficiency attenuates the methamphetamine-induced increase in ASIC1α and ASIC2,but aggravates neuronal injury and cognitive dysfunction in the prefrontal cortex.关键词
甲基苯丙胺/核因子E2相关因子2/酸敏感离子通道1α/酸敏感离子通道2/前额叶皮质/神经元损伤Key words
methamphetamine/nuclear factor erythroid 2-related factor 2/acid-sensing ion channel 1α/acid-sensing ion channel 2/prefrontal cortex/neuronal injury分类
医药卫生引用本文复制引用
普珊,孙成会,许睿珂,王敏,李利华,洪仕君,杨根梦..Nrf2缺失影响甲基苯丙胺致前额叶皮质ASIC表达及神经元损伤[J].中国药理学与毒理学杂志,2026,40(7):481-492,12.基金项目
国家自然科学基金项目(82202081) （82202081）
国家自然科学基金项目(82260336) （82260336）
国家自然科学基金项目(82371898) （82371898）
国家自然科学基金项目(82560336) （82560336）
云南省科技厅基础研究项目(202301AT070269) （202301AT070269）
云南省科技厅基础研究项目(202301AU070216) （202301AU070216）
云南省科技厅-昆医联合专项(202301AY070001-234) （202301AY070001-234）
云南省科技厅-昆医联合专项(202501AY070001-002) （202501AY070001-002）
昆明医科大学法医毒物学一流学科团队(2024XKTDYS10) National Natural Science Foundation of China(82202081) （2024XKTDYS10）
National Natural Science Foundation of China(82260336) （82260336）
National Natural Science Foundation of China(82371898) （82371898）
National Natural Science Foundation of China(82560336) （82560336）
Yunnan Fundamental Research Projects(202301AT070269) （202301AT070269）
Yunnan Fundamental Research Proj-ects(202301AU070216) （202301AU070216）
Yunnan Provincial Department of Science and Technology-Kunming Medical University Joint Special Project(202301AY070001-234) （202301AY070001-234）
Yunnan Provincial Department of Science and Technology-Kunming Medical University Joint Special Project(202501AY070001-002) （202501AY070001-002）
and First-Class Discipline Team of Forensic Toxicology,Kun-ming Medical University(2024XKTDYS10) （2024XKTDYS10）
