摘要： 目的 探讨二十二碳六烯酸（DHA）对甲基苯丙胺（METH）诱导的人神经母细胞瘤SH-SY5Y细胞神经毒性的作用及其作用机制。方法 体外培养SH-SY5Y细胞，分为Control组、METH组以及METH+DHA组。显微镜下观察SH-SY5Y细胞形态，测量并计算细胞长轴和短轴的比值，通过流式细胞术检测各组SH-SY5Y细胞凋亡水平。采用蛋白质组学分析METH引起的SH-SY5Y蛋白组改变，并筛选加入DHA后表达回调的蛋白。结果 与Control组比较，METH组SH-SY5Y细胞神经突明显短缩，长轴与短轴比值减小（均P < 0.05）；与METH组比较，METH+DHA组神经突伸长，细胞长轴与短轴比值增大（均P < 0.05）。与Control组比较，METH组早期细胞凋亡和晚期细胞凋亡比例升高，且总凋亡细胞比例也明显升高（均P < 0.05）；与METH组比较，METH+DHA组晚期细胞凋亡和总凋亡细胞比例降低（均P < 0.05）。蛋白组学分析显示，加入DHA可以将22个METH诱导下表达变化的蛋白反向调节，涉及了核糖体、细胞骨架和谷胱甘肽代谢等生物学功能。结论 DHA可减弱METH诱导的SH-SY5Y细胞神经毒性，其机制可能与DHA调控角蛋白表达，改善谷胱甘肽代谢等信号通路有关。

二十二碳六烯酸对甲基苯丙胺诱导的人神经母细胞瘤细胞神经毒性的影响

陈芳萍¹, 王琪¹, 郑丹², 官志忠³, 楼迪栋^1,4

1. 贵州医科大学法医学院法医病理学教研室, 贵阳 550004;
2. 贵阳市妇幼保健院围产期保健科, 贵阳 550003;
3. 贵州医科大学地方病与少数民族疾病教育部重点实验室, 贵州省医学分子生物学重点实验室, 贵阳 550004;
4. 贵州中医药大学基础医学院法医学教研室, 贵阳 550025

收稿日期:2020-11-16出版日期:2021-10-30发布日期:2021-10-11
通讯作者:楼迪栋E-mail:15761879@qq.com
作者简介:陈芳萍(1995-),女,硕士研究生.
基金资助:国家自然科学基金（81760336）


关键词: 二十二碳六烯酸, 甲基苯丙胺, SH-SY5Y细胞, 蛋白质组学
Abstract: Objective To evaluate the effect of docosahexaenoic acid (DHA) on neurotoxicity induced by methamphetamine (METH) in human neuroblastoma SH-SY5Y cells and elucidate the underlying mechanism. Methods SH-SY5Y cells were cultured in vitro and divided into the Control group, METH group, and METH +DHA group according to treatment. The morphology of SH-SY5Y cells in each group was observed under a microscope, and the ratio of the long axis to the short axis of SH-SY5Y cells was determined. Apoptosis of SH-SY5Y cells was determined by flow cytometry. Proteomic changes in SH-SY5Y cells caused by METH were analyzed, and the proteins reversely regulated after DHA addition were screened. Results The neurites of SH-SY5Y cells were significantly shortened and the ratio of the long axis to the short axis was decreased in METH group compared to those in the Control group (P < 0.05). Compared to METH-treated cells, METH+DHA treated cells had elongated neurites and an increased ratio of the long axis to short axis (P < 0.05). The proportion of early cell apoptosis and late cell apoptosis was increased and the proportion of total cell apoptosis was also significantly increased in the METH group compared to those in the Control group (P < 0.05). The proportions of late cell apoptosis and total cell apoptosis were decreased in the METH+DHA group compared to those in the METH group (P < 0.05). Proteomic analysis revealed that the addition of DHA could reversely regulate 22 METH-altered proteins involved in biological functions such as ribosome, cytoskeleton, and glutathione metabolism. Conclusion DHA can reduce METH-induced neurotoxicity in SH-SY5Y cells, and its mechanism may be related to the regulation of keratin expression, improvement of glutathione metabolism, and other signaling pathways.
Key words: docosahexaenoic acid, methamphetamine, SH-SY5Y cells, proteomics
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