| 摘要: |
| 【目的】研究H3N2和H1N1甲型流感病毒感染对仔猪肺脏CDK9基因表达的影响，揭示流感发病机制与CDK9基因的相关性。【方法】以接种PBS的仔猪为空白对照，利用实时荧光定量PCR技术和半定量免疫组织化学法，检测H3N2和H1N1甲型流感病毒接种感染7 d后，仔猪肺脏CDK9的mRNA和蛋白表达的变化，并在电子显微镜下观察CDK9阳性细胞的定位。【结果】实时荧光定量PCR检测表明，H3N2和H1N1甲型流感病毒感染仔猪7 d后，肺脏组织内CDK9基因mRNA的相对表达量分别为0.42和0.36，与对照（1.0）相比显著降低(P＜0.05)。半定量免疫组织化学检测结果显示，仔猪受到H3N2和H1N1流感病毒感染后，肺脏组织内CDK9蛋白的相对表达水平分别为31.4和38.7，较正常组织（61.4）显著降低(P＜0.05)。电子显微镜观察发现，H3N2和H1N1甲型流感病毒感染仔猪后，肺脏细胞界限不明显，CDK9阳性细胞散在分布于终末细支气管和肺泡内，且胞核着色较深，但是细胞整体着色变浅。【结论】受到流感病毒感染后，仔猪肺脏组织内CDK9表达量降低，可能是RNApolⅡ磷酸化水平降低的原因之一。 |
| 关键词: 流感病毒 CDK9基因 实时荧光定量PCR 免疫组织化学 |
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| 基金项目:国家自然科学基金项目(31072115) |
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| Effect of H3N2 and H1N1 influenza A virus infection on expression of porcine CDK9 gene |
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| Abstract: |
| 【Objective】This study aimed to investigate the influence of influenza A virus H3N2 and H1N1 infection on the expression of CDK9,and uncover the association between pathogenesis of influenza A virus and CDK9.【Method】qRT-PCR and semi-quantitative immunohistochemical assay were applied to detect the mRNA and protein expression levels of CDK9 in swine lung tissue 7 d after infection of H3N2 and H1N1 virus.Locations of CDK9 positive cells were observed with electron microscope.Piglets inoculated with PBS were used as blank control.【Result】qRT PCR detection showed that CDK9 mRNA expression levels were 0.42 and 0.36 after being infected with influenza A virus H3N2 and H1N1,respectively,which were significantly reduced (P＜0.05) compared with blank control (1.0).Semi-quantitative immunohistochemistry assay showed that the expression of CDK9 protein in piglet tissues infected with H3N2 and H1N1 decreased to 31.4 and 38.7 from 61.4 (P＜0.05).Electron microscope observation showed that lung cell boundaries became vague after influenza A virus H3N2 and H1N1 infection,CDK9-positive cells scattered in the terminal bronchiole and alveolus,and the nucleus stained dark,while the overall color of cells became shallow.【Conclusion】The expression of porcine CDK9 decreased after influenza A virus infection,which might be associated with the decreased of RNApolⅡ phosphorylation level after influenza A virus infection. |
| Key words: influenza virus CDK9 gene qRT-PCR immunohistochemistry |
