利血平脾虚证模型海马基因表达谱变化的研究

Primary study on changes of gene expression profile in hippocampi of reserpine animal models with spleen-asthenia

目的 应用基因芯片技术研究利血平脾虚证大鼠模型海马基因表达谱的变化。方法 雌性Wistar大鼠 3 0只 ,按体质量均衡随机分为实验组和对照组 ,实验组用利血平注射液背部皮下注射 ,造模第 1～ 9天 0 .15mg/ (kg·d) ,第 10～ 12天 0 .2 5mg/ (kg·d) ,第 13天 0 .5mg/(kg·d)。对照组同法注射同量注射用水。两组均第 14天处死 ,取海马 ,提取总RNA ,逆转录为cDNA ,用Cy3和Cy5两种荧光物质分别标记两组大鼠海马组织的cDNA ,制成探针以备检测。选用 2张BioDoorChip 40 96基因表达谱芯片 ,将探针与芯片杂交 ,计算机扫描后 ,用GenePixPro3 .0软件进行分析。结果 筛选出 2张芯片上 40 96条靶基因中 ,与脾虚证模型基因差异表达一致的基因 51条 ,占检测基因的 1.2 5% ,其中表达下调的基因 3 7条 ,已知基因 8条 ;表达上调的基因 14条 ,已知基因6条。模型大鼠海马的基因表达改变涉及免疫、炎症反应、蛋白聚糖、细胞骨架、离子通道、信号传导等多方面。结论脾虚证实质涉及海马的基因表达谱改变 。

Objective:It is to study the changes of gene expr ession profile in hi ppocampi of reserpine rat models with spleen-asthenia with gene CMOS chip techn i que. Methods:30 Wistar rats which were all female were randomly divided into ex periment group and control group according to balance body mass. The experiment group were injected reserpine injection under subcutaneous dorsum, which were us ed 0.15 mg/(kg·day) from the first day to the ninth day,0.25 mg/(kg·day) in the next three days and 0.5 mg/(kg·day) on the 14th day. The control group w e re injected the same dose physiological saline with the same method. The rats we re killed on the 14th day. The hippocampi of rats were taken out to extract gene ral RNA for reverse transcription cDNA. The cDNA in the two groups were marked w ith Cy5 and Cy3 fluorescence respectively as probes. The probes were hybridized with the two pieces of BioDoor Chip 4?096 gene expression profile CMOS chip and s canned with computer. Then the data were analyzed with GenePix Pro 3.0 software. Results:There were 51(1 25%) genes with altered expression am ong the 4?096 tar get genes. There were 8 known-genes among 14 up-regulated genes and 6 known-g ene s among 37 down-regulated genes. The changes were involved in the function of i m munity, Inflammatory reaction, proteoglycan, cytoskeleton, ion channel and signa l conduction etc. Conclusion:Parenchyma of spleen-asthenia is involved in the c hanges of gene expression profile in hippocampi, but the alter is fewer than in cerebral cortex.