研究氦氧饱和高气压暴露对铜绿假单胞菌基因表达谱的系统影响,对急性毒力基因表达的调节.用全基因组DNA芯片分析技术比较菌株暴露前后基因表达谱差异;RT-PCR方法验证部分差异表达基因;用分光光度法在细胞水平验证弹性蛋白酶含量;小鼠染毒法观察暴露组细菌毒力在整体动物水平的变化.基因表达谱分析结果表明,铜绿假单胞菌暴露12h差异表达基因达243个、72 h差异表达基因为1 168个.72 h差异表达基因中与细菌应激响应、蛋白折叠、转录调节、菌毛和鞭毛合成、毒力因子调节与合成、细菌外膜蛋白和抗原合成的基因大量上调;部分基因的RT-PCR验证结果与芯片结果一致;细胞水平验证结果显示暴露72 h细菌毒力表型增强.因此,氦氧饱和高气压暴露对铜绿假单胞菌基因表达谱有明显影响,对急性侵袭性感染毒力因子基因表达水平有正向诱导调节作用.%To elucidate the systemic gene expression variation and virulence gene regulation of Pseu-domonas aeruginosa in hyperbaric oxyhelium exposure environment. The level of differential gene expression was analyzed by PAO1 genomic DNA chip and 5 strikingly up-regulated genes were confirmed by RT-PCR. The level of elastase was detected by spectrophoto-metric method. Microarrays analysis showed that the expression levels of 243 genes were altered after 12 hours exposure and 1 168 genes altered after 72 hours exposure under the hyperbaric oxyhelium condition. These genes aremainly related to stress-sense/response, protein folding, transcriptional regulation, pili and flagellum metabolism, virulence factors adaptation, membrane proteins or antigens metabolism. The results of RT-PCR showed the same expression trends as in the micro-array. The ability of bacterial infection also increased after 72 hours exposure. Based on the results, we concluded that the hyperbaric oxyhelium environments affected PAO1 genes expression and several virulence-related genes were up-regulated.
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