Page 51 - 南京医科大学学报自然科学版

第42卷第8期 南京医科大学学报（自然科学版）
2022年8月 Journal of Nanjing Medical University（Natural Sciences） ·1093 ·

·基础研究·

甲基苯丙胺的神经毒性：基于蛋白组学的靶向神经突触损伤研究

彭 鹏 ，解翠薇 ，王 军 ，肖 杭 2
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2
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1 南京卫生高等职业技术学校临床一教研室，江苏 南京 210038；南京医科大学公共卫生学院毒理学教研室，江苏 南京
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211166

［摘 要］ 目的：探讨甲基苯丙胺（methamphetamine，METH）暴露后原代神经元蛋白组学及其对细胞功能的影响，整体阐述
METH的神经毒性及其影响通路。方法：将原代培养的神经元经METH（0、900 μmol/L）处理，蛋白酶解后，利用液相色谱⁃串联
质谱（LC⁃MS/MS）对其进行检测，利用Uniprot_RattusNorvegicus_36080_20180123数据库对其进行鉴定，利用基因本体论（gene
ontology，GO）分析进行蛋白功能注释，采用京都基因与基因组百科全书（Kyoto Encyclopedia of Genes and Genomes，KEGG）数据
库分析将目标蛋白序列进行归类，获取目标蛋白参与的信号通路；基于上述通路，利用Western blot验证METH引起的神经元
突触数量及表达改变；通过免疫荧光法观察突触间的表达及空间定位。结果：共鉴定出蛋白总数为 6 619个。与对照组比较，
表达差异（上调/下调）倍数大于1.2倍，且P < 0.05的蛋白质归为差异蛋白，通过筛选，METH处理组与对照组的差异蛋白有51种，
其中上调的蛋白40种，下调的蛋白11种。蛋白表达方面，METH 暴露显著降低突触后标志物PSD95及棘突标志蛋白Drebrin
的表达，而突触前标志物Synapsin1明显上调；免疫荧光结果显示，Drebrin表达明显下调且与F⁃actin的共定位减少，而PSD95与
Synapsin 1的空间定位亦显著下降。结论：METH暴露可引起原代神经元多种蛋白表达异常，细胞功能发生紊乱，突触结构破
坏，引发神经元损伤。
［关键词］ 原代神经元；蛋白组学；神经突触；神经损伤
［中图分类号］ R392.12 ［文献标志码］ A ［文章编号］ 1007⁃4368（2022）08⁃1093⁃06
doi：10.7655/NYDXBNS20220807

Neurotoxicity of methamphetamine：a proteomic study of targeted synaptic damage
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PENG Peng ，XIE Cuiwei ，WANG Jun ，XIAO Hang 2
1 Clinical Teaching Department of Nanjing Vocational Health College，Nanjing 210038；Department of Toxicology，
2
School of Public Health，Nanjing Medical University，Nanjing 211166，China
［Abstract］ Objective：To investigate the proteomics of primary neurons exposed to methamphetamine（METH）and the
corresponding cellular functions，unraveling the potential neurotoxicity of METH exposure. Methods：The primary cultured neurons
were treated with METH（0，900 μmol/L）and then hydrolyzed by protease. LC⁃MS/MS was used to detect and quantify the proteins，
then the database of Uniprot RattusNorvegicus_ 36080_ 20180123 was used to identify the target protein，gene ontology（GO）was used
to annotate the protein function，and Kyoto Encyclopedia of Genes and Genomes（KEGG）database was used to classify the target
protein sequence by KO analysis to obtain the target protein associated signal pathways. Based on the above pathways，Western blot
was used to verify the synaptic damage induced by METH，and immunofluorescence was performed to examine the expression and
spatial localization of synapses. Results：The total number of proteins identified was 6619. Compared with the control group，the
protein with expression difference more than 1.2 times and P < 0.05 was classified as differential protein. Through screening，51 different
proteins were screened，of which 40 proteins were up⁃regulated while 11 proteins were down⁃regulated. For protein expression，METH
exposure significantly decreased the expression of post synaptic density protein 95（PSD95）and Drebrin，while presynaptic marker
synapsin 1 was significantly up ⁃ regulated；immunofluorescence results showed that Drebrin expression was significantly decreased
accompanied by reduced co⁃ localization with F⁃actin，meanwhile，the spatial localization of PSD95 and synapsin 1 was significantly
decreased. Conclusion：METH exposure causes abnormal expression of numerous proteins in primary neurons as well as the disorder

［基金项目］ 江苏省自然科学基金（BK20191349）
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通信作者（Corresponding author），E⁃mail：601230640@qq.com