摘要
目的:研究大剂量水杨酸钠耳毒性的作用机制。方法:运用寡核苷酸基因芯片技术分析大剂量水杨酸钠注射对大鼠耳蜗基因表达谱的影响。联用TRIzol和RNA纯化柱分别从13周正常大鼠和水杨酸钠注射后的大鼠耳蜗内容物提取总RNA,将总RNA逆转录为cDNA,再将cDNA体外转录为cRNA,在此过程中同时进行生物素标记,将cRNA片段化后作为探针与质量检测芯片杂交,确定无问题后再与寡核苷酸基因芯片杂交扫描,用Affymetrix Microarray Suite software 5.0分析处理扫描结果,获得两种情况下耳蜗的基因表达谱。结果:表达上调2倍以上的基因和EST有42个,下调2倍以上的有49个。结论:水杨酸钠影响各种离子通道和突触蛋白基因,可能有助于解释水杨酸钠所致的耳聋和耳鸣作用机制,而其他变化基因亦有助于解释水杨酸钠广泛的药理作用。
OBJECTIVE To study the mechanism of ototoxicity of salycilate. METHODS Total RNAs from cochleas of normal rats and salicylate injected rats were extracted combination TRIzol with RNeasy column. Double-stranded eDNA were synthesized by the reverse transcription of the total RNAs, biotin-labeled cRNAs were prepared from eDNA by in vitro transcription then further fraigmented for target preparation. After an aliquot was analyzed by agarose gel electrophoresis, part of cRNAs were hybridized with test chip, the remaining target preparation were hybridized with the real chip. The hybridized chips were scanned and analyzed by Affymetrix Microarray Suite software 5.0. RESULTS Two type of gene expression profiles of cochleas suggested that 42 genes upregulated and 59 genes down-regulated over two times, which involved in sigal transduction, gene transcription regulation, ion channels, synaptie proteins, calcium binding proteins, oxidation reduction, et al. CONCLUSlON Evident gene expression changes of Scn8a and other types of ion channels and synaptic proteins genes have been found, in which Scn8a is sodium ion channel gene and associated with auditory function. This may be contributed to interpret the mechanism ofototoxicity of salicylate-hearing loss and tinnitus. Furthermore the other gene expression changes may help to underlie the broad pharmacologic effects of salicylatel.
出处
《中国医院药学杂志》
CAS
CSCD
北大核心
2007年第12期1659-1662,共4页
Chinese Journal of Hospital Pharmacy
关键词
基因芯片
基因表达谱水杨酸钠
耳毒性
gene chip
gene expression profile
sodium salicylate
ototoxicity
