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目的 利用基因表达芯片检测反复摄取烟碱对大鼠脑内膜受体和细胞信号转导系统相关的蛋白激酶基因表达的影响。方法 大鼠每天 2次皮下注射烟碱 (1 2mg·kg-1) ,连续用药 2wk后取全脑 ,提取RNA ,逆转录合成cD NA ,转录合成生物素化RNA ,并将其片断化后与芯片杂交 ,对荧光信号扫描分析。结合RT PCR方法对芯片分析结果进行验证。结果 反复摄入烟碱 ,脑内共有 2 2 4个重要蛋白的基因表达发生变化。其中 ,蛋白激酶、膜受体和离子通道共 94个发生变化。在蛋白激酶中 ,MAPK激酶等 13种上调 ;IP3 激酶等 7种发生下调。在膜受体中 ,NMDA受体等 9种发生上调 ;GluRD等 13种发生下调。结论 反复给予烟碱可影响多种膜受体和细胞信号转导相关基因的表达水平 ,这些变化可能与脑内N受体失敏有关。
Objective To detect the effect of nicotine on protein kinase gene expression related to rat brain endoplasmic reticulum receptor and cell signal transduction system by gene expression microarray. Methods Rats were injected subcutaneously twice daily with nicotine (12 mg · kg -1) for 2 weeks. The whole brain was harvested for RNA extraction and cD NA was reverse transcribed to synthesize biotinylated RNA. Chip hybridization, fluorescence signal scanning analysis. Combined with RT PCR method to verify the results of chip analysis. The results of repeated intake of nicotine, brain a total of 224 kinds of important protein gene expression changes. Among them, 94 protein kinases, membrane receptors and ion channels changed. Among protein kinases, MAPK kinase and other 13 kinds of up-regulation; IP3 kinase and other 7 kinds of down-regulation. Among membrane receptors, 9 such as NMDA receptors were upregulated; 13 such as GluRD were down-regulated. Conclusions Nicotine can affect the expression of various membrane receptors and cell signal transduction related genes, which may be related to the desensitization of N receptor in the brain.