【摘要】 目的:研究多聚腺苷二磷酸核糖聚合酶（poly（ADP-ribose）polymerase,PARP）抑制剂PJ34对绿脓菌素（Pyocyanin,PCN）引起的腹腔来源巨噬细胞RAW264.7损伤的保护作用。方法:分别采用800nM的PJ34预处理细胞2h（对照组给与0.1% Dimethyl sulfoxide,DMSO）后,不同浓度的PCN（50μM,100μM）刺激24h,MTT法检测绿脓菌素及PJ34对RAW264.7细胞存活率的影响;采用800nM的PJ34预处理细胞2h（对照组给与0.1%DMSO）,给与PCN（100μM,3h）刺激,荧光显微镜及流式细胞术检测细胞内活性氧自由基（Reactive oxidant species,ROS）的水平,并采用激光共聚焦显微镜观察RAW264.7细胞内钙离子水平;或加入18μg·mL^-1碳颗粒继续共孵育2h,显微镜下观察RAW264.7细胞吞噬碳颗粒的情况。结果:PJ34预处理组细胞与单用PCN处理组比较,细胞存活率提高近20%;PJ34预处理缓解了PCN刺激诱导的RAW264.7细胞内ROS和Ca²⁺水平的升高（P<0.05）,并降低了RAW264.7细胞由PCN刺激引起的吞噬功能损伤。结论:PJ34通过抑制PCN引起的RAW264.7细胞内ROS和Ca²⁺水平升高,提高细胞的存活率并增强细胞的吞噬功能。更多还原

【Abstract】 Objective:To investigate the protective effects of poly ADP-ribose polymerase（PARP）inhibitor PJ34 in the pyocyanin（PCN）induced dysfunction of RAW264.7 cells.Methods:MTT assay was employed to measure the cell viability of RAW264.7 at 24 hpost-PCN（50μM,100μM）challenge following 2 hpre-treatment of PJ34 or 0.1% DMSO.ROS generated in RAW264.7 pre-treated with PJ34 or 0.1% DMSO for 2 hprior to 3 hof exposure to 100μMPCN was observed by DHE staining and quantified by Flow cytometry.The Ca²⁺level generated in RAW264.7 cells with the same treatment was observed by the laser confocal microscopy.Phagocytosis of carbon particles in PCN-stressed RAW264.7 cells with/without pre-treatment of PJ34 was observed under fluorescence microscopy and analyzed by image J.Results:Compared with the un-pretreated group,the cell viability of PJ34 pretreatment group was enhanced by nearly 20% under PCN stimulation.PJ34 pretreatment attenuated the increase of ROS and Ca²⁺levels in RAW264.7 cells induced by PCN stimulation（P<0.05）.Carbon particles with RAW264.7 cells under PCN-challenge were significantly decreased and pre-treatment of PJ34 reversed such decrement.Conclusion:PCN induced oxidative damage of RAW264.7 macrophages in a dose-dependent manner.PJ34 shows protective effects on oxidative damage of RAW264.7 cells induced by PCN.更多还原