实验性自身免疫性脑脊髓炎小鼠模型的基因芯片研究

Gene expression profile in mice model of experimental autoimmune encephalomyelitis using cDNA microarrays

目的 用 4 0 96点基因芯片筛选实验性自身免疫性脑脊髓炎小鼠模型有表达差异的基因。方法 将 4 0 96条基因cDNA产物按微矩阵排列点样在玻片上。以髓鞘蛋白脂质蛋白多肽139 15 1及福氏佐剂为抗原免疫SJL小鼠 ,取免疫后 3～ 4周急性发病的小鼠及同代正常小鼠各 6只 ,每一只急性发病的小鼠及其对照为一组 ,分别取全脑组织 ,于 - 80℃冰箱保存。提取小鼠全脑组织总RNA ,RT PCR逆转录成cDNA。标记cDNA探针 ,分别用cy3 dUTP、cy5 dUTP标记正常小鼠和实验性自身免疫性脑脊髓炎小鼠的cDNA。将基因芯片和杂交探针变性后杂交 ,洗干。用ScanArray4 0 0 0扫描芯片 ,GenePixPro 3 0软件分析cy3、cy5两种荧光信号的强度和比值。结果 1～ 6组中筛选出差异表达的数据分别为 4 3、30、176、76、2 94和 12 9项。这些基因在与两种探针杂交时表现出一定的差异。 6个组中一致发现表达异常的基因涉及了免疫相关基因、细胞骨架蛋白基因、细胞周期相关基因、能量代谢相关基因、蛋白质合成相关基因、信号转导及离子通道相关基因等广泛的基因变化。结论 除免疫相关基因外 ,细胞骨架蛋白相关基因、能量代谢相关基因、蛋白质合成相关基因、细胞周期相关基因、离子通道相关基因等可能参与了自身免疫性脑脊髓膜炎的发病。

Objective To construct the genes expression profile database of experimental autoimmune encephalomyelitis (EAE) mice model and to screen the high candidate genes with microarray analysis. Methods Polymerase chain reaction (PCR) products on behalf of 4 096 genes from mixed mice tissue library were assembled on the modified Oako glass slide. The general RNA from 6 EAE mice model and 6 normal mice head was transcripted into cDNA by RT-PCR and labelled with cy-3 and cy-5.6 groups each consist one EAE mice and a control. These two probes were mixed and hybridized with the above mentioned gene chips. Scan array 4000 was used for scanning the hybridizing signals and GenePixPro 3.0 for date analysis. Results Of the total 4 096 genes monitored, 43 genes in group one showed changes in expression level of the ratio outside 0.5 and 2, 30 genes differed in group two, 176 genes changed in group three, 76 in group four, 294 in group five and 129 in group six. Conclusions The results showed the consistent different genes expression throughout the EAE mice model. The genes related to immune, cell structure, cell cycle, ion channel, signal transduction, protein synthesis and metabolism are involved in EAE pathogenesis. The results provide deeper insights into the mechanism of EAE and multiple sclerosis.