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微生物学通报

呼吸道合胞病毒感染人肺上皮细胞一氧化氮的产生及其作用
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安徽省高等学校“十五人才”优秀中青年骨干教师基金(No. 安徽省教育厅教人[2002] 023号)


The Production and Effect of Nitric Oxide in Respiratory Syncytial Virus-infected Human Lung Epithelial Cells
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    摘要:

    本文探讨呼吸道合胞病毒(RSV)感染人肺上皮细胞(A549细胞)后, 一氧化氮(NO)的水平变化及其在RSV感染中的氧化损伤和抗病毒作用。RSV以不同时间感染A549细胞, 并给予NO合成的抑制剂氨基胍(AG)处理。收集细胞培养上清, 分别用硝酸还原酶法和硫代巴比妥酸法检测NO和丙二醛(MDA)含量, 化学法检测羟自由基(OH·)与超氧阴离子(O2.—)水平, 空斑形成试验测定病毒复制滴度(PFU)。结果显示在RSV感染4 h后即上调NO、OH·、O2.—和MDA的表达水平。当RSV感染中给予AG处理以抑制iNOS合成NO时, 则降低OH·、O2.—和MDA含量, 但病毒PFU升高。各指标的变化与相应时间点的感染组相比, 差异均有显著性。提示RSV感染肺上皮细胞诱导生成的NO与细胞内自由基水平升高和加重细胞的自由基损伤程度有关; 但在一定程度上可抑制病毒的增殖水平。

    Abstract:

    To investigate the formation and effect of nitric oxide (NO) on oxidative injury and anti-virus in human lung epithelial cells (A549) infected with respiratory syncytial virus (RSV) in vitro. A549 cells infected with RSV treated or untreated with Aminoguanidine (AG), the specific inhibitor for inducible nitric oxide synthase, were used to collect cellular supernatants and do the experiments. At indicated time points, the concentrations of NO, hydroxy radical (OH·), superoxide anion (O2.—) and malondialdehyde (MDA) were measured according to the kit reference. The viral titers were evaluated by plaque forming unit (PFU) assay. We found that RSV infection could markedly up-regulate the levels of OH· and O2.— in A549 cellular supernatants since 4 hours after infection. Meanwhile, the levels of MDA increased obviously in a time-dependent manner. The changes of each index varied significantly compared with the control group. But the NO content in the cellular supernatants could significantly decrease when AG inhibitor was used, and the levels of OH·, O2.— and MDA were also decreased accordingly in comparison with the corresponding RSV-infected group. In addition, the titer of virus was increased, which was about 1.5 times as many the RSV group 24 h after RSV infection. The changes of these indices have significantly different compared with the corresponding RSV-infected group. In conclusion, RSV infection could induce high-level NO. The NO may associate with the content of free radical, which could decrease the viral titers in the infection, but lead oxidative injury to normal cells.

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黄升海,王 杨,刘 伟,史晓佾. 呼吸道合胞病毒感染人肺上皮细胞一氧化氮的产生及其作用[J]. 微生物学通报, 2009, 36(5): 0735-0740

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