摘要
目的利用基因芯片技术研究藜芦定碱对SH-SY5Y细胞相关基因表达的影响。方法体外培养神经细胞SH-SY5Y,不同浓度的藜芦定碱作用8 h后,提取细胞总RNA,纯化mRNA后逆转录制备目标序列,与含有29000种人cD-NA的表达谱芯片HOA 5.1杂交,以AXON4000B荧光扫描仪扫描芯片上的荧光信号,获得的荧光信号的强度应用Rosetta ResolverSystem软件进行差异表达谱分析。结果与正常对照组相比,SH-SY5Y细胞受藜芦定碱50,200,800μmol.L-1作用后,显著差异表达基因分别有71,182,77条,其中上调基因分别有45,81,45条,下调基因分别为26,101,32条。差异表达基因涉及细胞物质与能量代谢、线粒体功能相关基因,细胞周期、细胞分化、细胞凋亡及细胞信号传导相关基因。结论采用基因芯片技术筛选藜芦定碱对SH-SY5Y细胞毒性相关基因,寻找藜芦定碱毒性作用的靶基因,可能为藜芦定碱对神经毒性机制研究提供新思路。
Aim To screen the differently expressed genes of SH-SYSY cell treated with veratridine by using cDNA microarray. Methods In vitro cultured SH-SY5Y cells treated with Veratridine for 8 hours, and total RNA was isolated. The target sequences were hybridized to cDNA microarray HOA 5.1 of 29000 human genes. After washing, the cDNA microarraies were scanned by fluorescent scanner AXON4000B. And the fluorescent signals were analyzed by Rosetta Resolver？ System. Result Compared with the control group, there were 71,182,77 differential expressed genes in SH-SY5Y treated by veratridine 50,200,800 μmol · L^-1. The differential expressed genes were related with Material and energy metabolism, mitochondrial function, the cell cycle, differentiation and apoptosis , cell signal conduct, et al. Conclusion The investigations based on cDNA microarray can sequence related gene in SH-SY5Y treated by veratridine,and may provide a new idea in studing the neurotoxieity of veratridine.
出处
《中国药理学通报》
CAS
CSCD
北大核心
2013年第5期643-647,共5页
Chinese Pharmacological Bulletin
基金
国家重点基础研究发展计划资助项目(No2011CB505304
2012CB518402)
