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目的利用基因芯片技术研究富氢水对γ射线照射后Beagle犬外周血淋巴细胞基因表达的影响。方法将雄性Beagle犬随机分为对照组、单纯照射组和富氢水组(n=5),采用基因芯片技术对2.0Gy 60 Coγ射线照射后6h各组Beagle犬外周血淋巴细胞的差异表达基因进行筛选,利用GO(gene ontology)和KEGG(Kyoto encyclopedia of genes and genomes)数据库对差异表达基因进行生物信息学分析,并应用实时荧光定量PCR技术对基因芯片结果进行验证。结果单纯照射组筛选出差异表达2倍以上的基因4 730条,富氢水组筛选出差异表达2倍以上的基因4 493条,单纯照射组与富氢水组共同差异表达2倍以上的基因有1 606条;单纯照射组的差异表达基因对应的生物学过程、细胞组分及分子功能分别有10、9、3个功能簇,富氢水组差异表达基因对应的生物学过程、细胞组分及分子功能分别有15、3、4个功能簇。单纯照射组差异表达基因涉及19条生物学通路,富氢水组差异表达基因涉及24条生物学通路,单纯照射组与富氢水组的差异表达基因有5条共同生物学通路。选择2条差异表达基因进行mRNA水平的验证,PCR扩增结果与芯片结果具有良好的一致性。结论富氢水对γ射线照射后Beagle犬外周血淋巴细胞基因表达的影响,可能涉及多种生物学过程、细胞组分、分子功能的改变及多条信号通路的激活。
Objective To study the effects of hydrogen-rich water on gene expression of peripheral blood lymphocytes in Beagle dogs after γ-ray irradiation by gene chip technology. Methods Male Beagle dogs were randomly divided into control group, irradiation group and hydrogen-rich water group (n = 5). The gene expression of differentially expressed genes in peripheral blood lymphocytes of Beagle dogs at 2.0 Gy 60Coγ ray irradiation The bioinformatics analysis of differentially expressed genes was carried out by using GO (gene ontology) and KEGG (Kyoto encyclopedia of genes and genomes) databases, and the real-time PCR method was used to verify the results of microarray. RESULTS: 4 730 genes with more than 2 fold differential expression were screened out in the irradiation alone group, 4 493 genes with more than 2 fold difference were screened in the hydrogen-rich water group, those with more than 2-fold difference in the group with hydrogen peroxide alone There were 1 606 lines. The biological processes, cell components and molecular functions corresponding to the differentially expressed genes in the group of irradiation alone had 10, 9, and 3 functional clusters, respectively. The biological processes corresponding to differentially expressed genes in hydrogen-rich water group, Points and molecular functions were 15,3,4 clusters. There were 19 biological pathways involved in the differentially expressed genes in pure irradiation group, 24 genes were involved in the differential expression genes in hydrogen-rich water group, and 5 common biological pathways were found in the differentially expressed genes in pure irradiation group and hydrogen-rich water group. Select two differentially expressed genes for mRNA level validation, PCR amplification results and chip results have good consistency. Conclusion The effect of hydrogen-rich water on the gene expression of peripheral blood lymphocytes in Beagle dogs after γ-ray irradiation may involve a variety of biological processes, changes in cellular components, molecular functions and activation of multiple signaling pathways.