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目的应用基因表达谱芯片筛选急性病毒性心肌炎相关基因,并对这些基因的功能进行初步分析。方法将8 464条小鼠PCR产物按微聚阵排列点样于化学涂层的载玻片上,制成基因芯片;用柯萨奇B3 (CVB3)病毒感染BALBc小鼠,在感染后的第4、8、21天,分别将正常和病毒性心肌炎心肌中mRNA逆转录,合成荧光分子(cy3／cy5)掺入的cDNA链制备表达谱探针,芯片杂交和严格洗片后,用ScanArray3000荧光扫描芯片荧光信号图像,利用计算机分析病毒性心肌炎心肌与正常心肌中丝氨酸苏氨酸磷酸化酶基因(PIM-3)、血管形成因子基因(Ang)-1、组织相容性基因-2的表达,并对获得的基因进行生物信息学分析。结果小鼠在CVB3感染后的第4、8、21天,心肌中分别发现了1 684、284、98条差异表达基因:PIM-3、Ang-1、组织相容性基因-2的表达被持续抑制。结论应用基因表达谱芯片可筛选出差异表达基因,PIM-3、Ang-1、组织相容性基因-2的差异表达可能与病毒性心肌炎发生和发展有关。
Objective To screen genes related to acute viral myocarditis by gene expression microarray and to analyze the function of these genes. METHODS: 8 464 mouse PCR products were microscopically arrayed on chemical-coated glass slides to make gene chips. BALBc mice were infected with Coxsackie B3 (CVB3) virus, , 8,21 days, reverse transcription of mRNA in normal and viral myocarditis myocardium, synthesis of fluorescent molecule (cy3 / cy5) cDNA chain preparation of expression profiling probe, chip hybridization and strict washing, scanning with ScanArray3000 fluorescence Chip fluorescence signal images, the expression of serine threonine phosphorylase gene (PIM-3), angiogenesis factor-1 (Angiopoietin-1) and histocompatibility gene-2 in myocardium and normal myocardium of viral myocarditis were analyzed by computer. The bioinformatics analysis of the obtained genes was carried out. Results The expression of 1 684,284,98 differentially expressed genes in myocardium was found on the 4th, 8th and 21st day after CVB3 infection, respectively. The expression of PIM-3, Ang-1 and histocompatibility gene-2 Continuous inhibition. Conclusion Gene expression profiling can differentiate differentially expressed genes. The differential expression of PIM-3, Ang-1 and histocompatibility gene-2 may be related to the occurrence and development of viral myocarditis.