富氢水对γ射线照射Beagle犬淋巴细胞基因表达谱的影响

Effect of hydrogen-rich water on lymphocyte gene expression profiles of Beagle dogs receiving gamma ray irradiation

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 effect of hydrogen-rich water on lymphocyte gene expression of Beagle dogs exposed to gamma ray （γ ray） using GeneChip technology. Methods Male Beagle dogs were divided into control group, irradiation （ IR） group and hydrogen-rich water treatment （HRW ＋ IR） group. GeneChip technology was applied to screen the differentially expressed genes in peripheral blood lymphocyte exposed to 2. 0 Gy ^60Co γ ray for 6 h. Gene ontology （GO） and Kyoto encyclopedia of gene and genomes （KEGG） databases were used to analyze the bioinformatics of the differentially expressed genes,and real-time quantitative PCR was used to validate the results of GeneChip. Results We screened 4 730 differentially expressed genes with over 2-fold changes in IR group and 4 493 genes in HRW＋IR group, and identified 1 606 co-differentially expressed genes in the IR and HRW＋IR groups. GO terms of biological process,cellular component and molecular function in the IR group were ten, nine and three, respectively, and in HRW＋IR group were fifteen, three and four,respectively. The differentially expressed genes were involved in 19 KEGG pathways in the IR group, 24 KEGG pathways in HRW＋IR group, and five KEGG pathways in both groups. The PCR results of two genes,selected to validate the GeneChip, were consistent with the GeneChip results. Conclusion Our results indicate that the effect of hydrogen-rich water on lymphocyte gene expression of γ ray-radiated Beagle dogs may involve changes of molecular function, cellular component and biological process,and activation of many signal pathways.