PARP-1抑制对PM2.5致人支气管上皮细胞炎症反应的保护作用

The Protective Effect of Inhibition of PARP-1 on Inflammation Induced by PM2.5 in Human Bronchial Epithelial Cell Line

目的 研究抑制多聚二磷酸腺苷核糖多聚酶-1（poly ADP-ribose polymerase-1,PARP-1）是否能缓解/逆转细颗粒物（PM2.5）诱导的炎症反应。方法 以不同质量浓度（0-1 000μg/mL）PM2.5处理正常人支气管上皮细胞（human bronchial epithelium cell line,HBE细胞）24h,台盼蓝拒染法测定细胞活力,采用200、400、600μg/mL PM2.5进行后续实验;设PM2.5（600μg/mL）单处理组、PARP-1抑制剂4-氨基-1,8-萘二胺（4-AN）（10μg/mL）单处理组、4-AN＋PM2.5组、溶剂（DMSO）对照组,免疫印迹法检测PARP-1、核因子κ-B（NF-κB）的p65亚基和诱导型一氧化氮合酶（iNOS）表达水平,硝酸酶还原法检测一氧化氮（nitric oxide,NO）水平。结果 PM2.5 200、400、600μg/mL单独处理HBE细胞时,细胞存活率随着PM2.5质量浓度增高而下降,PARP-1、p65核转位、iNOS和NO水平升高,400、600μg/mL PM2.5处理组与DMSO对照组比较,差异有统计学意义（P〈0.05）;4-AN预处理可拮抗PM2.5诱导的PARP-1表达与p65核转位升高,同时炎症介质NO与催化合成NO的iNOS水平显著下降（与PM2.5单处理组比较,差异有统计学意义,P〈0.05）,并恢复至正常水平（与4-AN单处理组和对照组比较差异无统计学意义,P〉0.05）。结论 抑制PARP-1可以明显缓解PM2.5对HBE细胞的致炎作用,其机制与下调NF-κB核转位进而阻断炎症介质表达有关。

Objective To explore whether the inhibition of poly ADP-ribose polymerase-1（PARP-1） could attenuated inflammation induced by fine particulate matter （PM2.5） in human bronchial epithelial cell line. Methods Cell viability was detected by Trypan Blue assay after incubated with PM2.5 for 24 h. PM2.5 doses no more than 600μg/mL were utilized in the following experiments. In order to observe how PARP-1 would effect the expression of nuclear factor-κB p65 and inducible nitric oxide synthase （iNOS）, cells were respectively treated with 600 μg/mL PM2.5, 10 μmol/L 4-amino-1, 8-naphthalimide （4-AN）, 600 μg/mL PM2. 5 ＋ 10 μmol/L 4-AN or DMSO. Western blot assay was used to estimate the protein expression of PARP-1, p65 in nuclear and iNOS in cytoplasm. Nitric acid enzyme reduction assay was used to determine the production of nitric oxide （NO）. Results As the PM2.5 concentration increased, the cell viability decreased, while the expression of PARP-1, p65, iNOS and NO increased significantly （P〈0.05）. After pretreatment of 4-AN for 24 h, the expression of PARP-1, p65, iNOS and NO almost decreased to the normal level （P〈0. 05）. Conclusion Inflammation triggered by PM2. 5 could be attenuated by the inhibition of PAPR-1, which involved the block of transcriptional activity of NF-κB for inflammatory mediator.