中文摘要：

目的观察补肺益肾方对单核细胞条件培养基刺激肺泡上皮细胞炎症反应的影响，探讨补肺益肾方治疗慢性阻塞性肺疾病（COPD）的抗炎机制。方法复制人髓系白血病单核细胞（THP-1）细胞株条件培养基刺激A549细胞炎症模型，分为空白对照组（20％空白兔血清）、模型组（20％空白兔血清＋25．0％THP-1细胞条件培养基）、核转录因子激活蛋白酶-1（AP-1）通路抑制剂T-5224组（20％空白兔血清＋25．0％THP-1细胞条件培养基4-100μmol／L T-5224）、补肺益肾组（20％含补肺益肾方兔血清＋25．0％THP-1细胞条件培养基）。用酶联免疫吸附试验（ELISA）检测细胞培养上清液中白细胞介素（IL-6、IL-8）、肿瘤坏死因子-α（TNF-α）、基质金属蛋白酶-9（MMP-9）等细胞因子的含量，用硫代巴比妥酸（TBA）法检测上清液中丙二醛（MDA）含量，用羟胺法检测总超氧化物歧化酶（T—SOD）活性，用凝胶迁移率实验（EMSA）检测AP-1通路的活性。结果与空白对照组比较，25．0％条件培养基作用24h、48h A549细胞增殖活性均明显增加[吸光度（A）值：24h为0．41±0．02比0．37±0．04，48h为1．30±0．09比1．15±0．19]；模型组IL-6、IL-8、TNF-α、MMP-9、MDA含量及AP-1表达均增多[IL-6（ng／L）：35．00±3．63比23．15±1．72，IL-8（ng／L）：273．09±164．36比231．45±33．90，TNF-α（ng／L）：51．61±9．51比28．87±3．34，MMP-9（ng／L）：442．85±78．86比235．60±14．62，MDA（μmol／L）：6．90±0．11比6．01±0．12，AP-1表达（A值）：2．260±0．062比1．000±0．000j。MDA/T—SOD比值增大（4．43±0．05比3．96±0．06）。与模型组比较，T一5224组IL-8（ng／L：100．29±17．03）、TNF—α（ng／L：25．13±0．46）、AP-1表达（A值：1．38±0．02）、MDA／T—SOD比值（4．23±0．23）和补肺益肾组MMP-9（ng/L：195．44±9．80）、MDA（μmol／L：5．86±0．30?

英文摘要：

Objective To observe the effects of nourishing lung and kidney formulas on inflammatory response of alveolar epithelial cells stimulated by monocytes conditioned medium and study the anti-inflammatory mechanism of the formulas for the treatment of chronic obstructive pulmonary diseases （COPD）. Methods The reproduction of inflammation models of A549 cells were stimulated by monocyte THP-1 cell strain conditioned medium. A549 cells were randomly divided into blank control group （20% blank rabbits serum）, model group （20% blank rabbits serum＋25.0% THP-1 cell conditioned medium）, nuclear transcription factor activator protein-1 （AP-1） pathway inhibitor T-5224 group （20% blank rabbits serum＋25.0% THP-1 cells conditioned medium＋100 μmol/L T-5224）, nourishing lung and kidney group （20% rabbits serum with nourishing lung and kidney formulas＋25.0% THP-1 cells conditioned medium）. The contents of interleukins （IL-6, IL-8）, tumor necrosis factor -α（TNF-α）, matrix metalloprotein-9 （MMP-9） in cell culture supernatant were detected with enzyme linked immunosorbent assay （ELISA）, the supernatant content of malondialdehyde （MDA） was detected with thibabituric acid （TBA） method, the total activity of superoxide dismutase （T-SOD） was detected with hydroxylamine method, and the activity of AP-1 pathway was detected with electrophoretic mobility shift assay （EMSA） method. Results Compared with the blank control group, the A549 cell proliferation were significantly increased at 24 hours, 48 hours stimulation by 25.0% cell conditioned medium （A value： 24 hours was 0.41 ±0.02 vs. 0.37 ±0.04, 48 hours was 1.30 ± 0.09 vs. 1.15 ±0.19）. Compared with the blank control group, the contents of IL-6, IL-8, TNF-α, MMP-9, MDA, AP-1 expression were significantly increased in model group [IL-6 （ng/L）： 35.00±3.63 vs. 23.15±1.72, IL-8 （ng/L）： 273.09± 164.36 vs. 231.45±33.90, TNF-α（ng/L）： 51.61 ±9.51 vs. 28.87±3.34, MMP-9 （ng/L）： 442.85±78.86