氧化应激环境下猪胸膜肺炎放线杆菌ohr基因转录活性的变化

Elevated transcription activity of ohr gene in Actinobacillus pleuropneumoniae under oxidative stress

使用异丙苯化过氧氢(Cumene hydroperoxide,CHP)模拟体内氧化应激环境,给予猪胸膜肺炎放线杆菌(Actinobacillμspleuropneμmoniae,APP)血清1型菌不同浓度CHP或相同浓度CHP不同时间的刺激,使细胞处于氧化应激状态,采用半定量RT-PCR方法,以看家基因recF为内参对照,观察ohr基因的转录活性,并进一步分析刺激强度、刺激时间与ohr基因转录活性的关系。结果表明,模拟的氧化应激环境明显增加了ohr基因的转录活性,该基因在转录水平上受到氧化应激调控。ohr mRNA的转录在一定范围内(CHP浓度不大于300μmol/L时)与氧化物刺激的强度具有时间依赖和浓度依赖关系。模拟的氧化应激促进了ohr基因的转录活性,由此可推知ohr基因可能是APP潜在的毒力基因,在APP致病过程中具有潜在的促进作用。

Stimulate Actinobacillus pleuropneuminiae serotype 1 ohr gene with different concentration of cumene hydroperoxide（CHP） or the same concentration of CHP at different times, through which mimicking the oxygen stress enviroment in the porcine, then confirm the transcription activity of ohr gene with semi-quantification RT- PCR, and using housekeeping gene recF as an internal standard. Analyze the correlation between the transcription activity of ohr gene with the strength of stimuli or the course. The oxygen stress enviroment,mediated by CHP,obviously upregulate the transcription activity of ohr gene and in a dose- and time-dependent manner, when expose to oxygen stress enviroment,the ohr mRNA level was obviously increased. Oxidative stress, mediated by CHP, can improve transcription activity of ohr gene, and ohr might be a potential virulence gene of Actinobacillus pleuropneumoniae ,it plays an important role in pathogenesis.