摘要
目的 研究抗抑郁剂相关基因 ,探讨其可能作用机制。方法与结果 以高浓度皮质酮 (Cort) 1 0μmol·L-1或Cort与三环类经典抗抑郁剂地昔帕明(DIM) 5 μmol·L-1共同处理PC1 2细胞 3d分别作为对照组和实验组 ,按Trizol一步法提取细胞总RNA并纯化mRNA ,将等量的对照组与实验组mRNA以逆转录法分别标记荧光素cy3和cy5 ,合成cDNA探针并等量混合 ,与小鼠 2 0 48点微矩阵基因芯片杂交 ,扫描荧光信号图像并分析比较二组差异表达基因 ,发现DIM与Cort共孵PC1 2细胞 3d可以诱导2 5 9个基因表达水平发生改变。其中 1 63个基因表达降低 ,如葡萄糖调节蛋白 78ku、葡萄糖激酶(GA)、细胞骨架蛋白、转化性生长因子 β受体结合蛋白、细胞色素C氧化酶、锂盐敏感性内消旋肌醇一磷酸酯酶等 ;有 96个基因表达水平升高 ,如神经元生长分化因子 9(GDF 9)、锌指蛋白 2 1 6、细胞色素P45 0、睾丸特异基因 1等。随机选取二个差异表达基因GA和GDF 9,以细胞原位杂交方法验证 ,取得与基因芯片评价一致的结果。结论 抗抑郁剂DIM作用可能与能量代谢、细胞骨架、营养因子、酶等多层次、多水平的基因表达改变有关。本研究利用基因芯片技术对抗抑郁剂相关基因进行了初步探索 。
AIM To study the antidepressant related genes and explore the possible action mechanism of antidepressants. METHODS and RESULTS PC12 cells were treated with corticosterone(Cort) 10 μmol·L -1 for 3 d in the absence or presence of desipramine(DIM) 5 μmol·L -1 and the mRNA was extracted, and then labeled with fluorescein cy3 or cy5 respectively using reverse transcription. The labeled probes were mixed equally and hybridized with the mouse 2048 gene dot microarray gene chip. After scanning and analysis, it was found that DIM induced 259 gene expression changes in Cort treated PC12 cells, in which 163 gene expression down regulated 〔for example, glucose regulated protein 78 ku, glucokinase (GA), cytoskeleton tubulin or actin, transforming growth factor β receptor binding protein, cytochrome C oxidase, lithium sensitive myo inositol monophosphatase A1, etc.〕 and 96 gene expression up regulated 〔for example, growth differentiation factor 9(GDF 9), zinc finger protein 216, cytochrome P450 subfamily Ⅳ B, testis specific gene 1, etc.〕. Using hybridization in situ, the effect of DIM on the GA or GDF 9 expression in Cort treated PC12 cells was also detected and the gene expression changes were consistent with results of gene chip detection. CONCLUSION It was indicated that the effect of DIM is associated with the gene expression changes at the levels of energy metabolism, cytoskeleton, neurotrophic factors, enzyme, ect.. The antidepressant related genes were elementarily studied with microarray gene chip and clues for further studies were provided.
出处
《中国药理学与毒理学杂志》
CAS
CSCD
北大核心
2003年第3期184-191,共8页
Chinese Journal of Pharmacology and Toxicology
基金
国家自然科学基金资助项目(39870876)~~
