NADPH氧化酶对HSV-1感染的小胶质细胞MMP-9激活的影响

NADPH oxidase activation is essential for HSV-1-induced MMP-9 expression in murine microgli

目的探讨NADPH氧化酶在单纯疱疹病毒1型（HSV．1）诱导的小鼠小胶质细胞（BV2）基质金属蛋白酶．9（MMP．9）表达中的作用。方法应用O．5、1．0mmol／LNADPH氧化酶特异性抑制剂Apocynin作用HSV-1诱导的BV2细胞，采用明胶酶谱分析细胞培养上清中MMP-9活性；RT—PCR检测NADPH氧化酶亚基p47phox及MMP-9mRNA的表达；Dihydroethidium（DHE）测定细胞内活性氧簇（ROS）的产生量。结果与正常对照组比较，HSV-1诱导的BV2培养上清中MMP-9活性增加，p47phox、MMP-9mRNA的表达增高，细胞内ROS水平增高约两倍以上，差异均有统计学意义（P〈0．05）；0．5mmoVLApocynin作用后可使上述变化减轻，但仍高于正常对照组，差异有统计学意义（P〈0．05）。结论NADPH氧化酶在HSV-1激活小胶质细胞MMP-9表达中可能起重要作用。

Objective To investigate the role of NADPH oxidase in Herpes simplex virus 1 （HSV-1）-induced matrix metalloproteinase 9 （MMP-9） expression in murine microglial BV2 cells. Methods BV2 cells induced by HSV-1 were divided into normal control group, HSV-1 infection group, and two apocynin treatment groups （in which apocynin was administered at 0.5 and 1.0 mmol/L after HSV-I infection）. MMP-9 gelatinolytic activity in the supernatants of the cell cultures was assessed by zymography. Semi-quantitative reverse transcription polymerase chain reaction （RT-PCR） was used to detect the mRNA expressions of NADPH oxidase subunit p47phox and matrix metalloproteinase-9 （MMP-9）, and the intraccllular reactivc oxygen species （ROS） levels were measured by dihydroethidium staining （DHE）. Results Compared to the normal control group, HSV-1 infection of the cells resulted in significantly up-regulated mRNA expression of NADPH oxidase subunit p47phox and MMP-9, and also in a 2-fold increase in the intracellular ROS level （P〈0.05）. These changes were attenuated by the application ofapocynin, but the mRNA expressions ofp47phox and MMP-9 and ROS level still remained significantly higher than those in the normal control group （P〈0.05）. Conclusion HSV-1 may induce MMP-9 activation through the generation ofNADPH oxidase-dependent ROS in murine microglia.