基于转录组测序技术揭示salsolinol的神经毒性作用  

Neurotoxic Effects of Salsolinol Revealing by Transcriptome Analysis

作　　者：张芳芳[1] 李梓晗 高怿宣 王雨桐[1] 冯予鑫 王佳楠 陈薛钗 ZHANG Fangfang;LI Zihan;GAO Yixuan;WANG Yutong;FENG Yuxin;WANG Jianan;CHEN Xuechai(Ministry of Environment and Life,Beijing University of Technology,Beijing 100124)

机构地区：[1]北京工业大学环境与生命学部,北京100124

出　　处：《生命科学仪器》2021年第6期29-34,共6页Life Science Instruments

基　　金：北京市自然科学基金面上项目(7202001);国家自然科学基金青年基金项目(81400935);北京市教委项目(KM202010005022)。

摘　　要：Salsolinol(1-甲基-6,7-二羟基-1,2,3,4-四氢异喹啉,Sal)是一种儿茶酚异喹啉类化合物,由多巴胺和醛类物质催化生成,其在神经系统中的毒性作用机制仍不清楚。本文主要探究Sal对细胞转录组的影响揭示其神经毒性的调控机制。方法选用大鼠肾上腺嗜铬细胞瘤PC12细胞进行Sal诱导,采用转录组测序分析(RNA-Seq)建立全转录组图谱,对差异基因进行GO分析和KEGG分析,最后采用Western-blot进行验证。结果 RNA-Seq分析Sal处理组和对照组的mRNA丰度差异,共鉴定出2405个显著性差异的表达基因(倍数变化≥2.0),包括740个上调基因,1665个下调基因。信号通路研究显示,这些基因与氧化应激、氧化磷酸化、自噬和溶酶体调节、PI3K-Akt信号通路、AMPK信号通路等密切相关,并发现Sal可以使Akt和mTOR蛋白的磷酸化水平增加。结论 Sal激活Akt-mTOR信号通路来调控细胞内的自噬水平,从而引发神经毒性作用。Salsolinol(1-methyl-6,7-dihydroxy-1,2,3,4-tetrahydroisoquinoline, Sal), one kind of catechol isoquinolines, is derived from dopamine and aldehydes. However, the neurotoxic effects of it are still remain unclear. Methods In this research, rat pheochromocytoma PC12 cells were used to analyze the cytotoxicity of Sal,and RNA-seq analysis was used to establish the whole-transcriptome profile to explore the differential abundance of mRNA. GO and KEGG analysis were carried out for differentially expressed genes(DEGs), and final confirmed by Western-blot. Results Comparative analysis of the mRNA abundance between control and Sal induced PC12 cells, a total of 2405 DEGs(multiple variation ≥2.0) with significant difference were identified, with the 740 unigenes up-regulated and 1665 unigenes down-regulated. By GO and KEGG analysis, these unigenes were found to be implicated in response to oxidative stress, oxidative phosphorylation, regulation of autophagy and lysosome,transferase activity, PI3 K-Akt signaling pathway, AMPK signaling pathway, etc. According to the result of WesternBlot, Sal was found to promote phosphorylation of Akt and mTOR. Conclusions Sal induces neurotoxicity by activating Akt-mTOR signaling pathways to regulate autophagy.

关 键 词：SALSOLINOL 转录组 差异表达基因 自噬 

分 类 号：Q189[生物学—神经生物学]