2023 年第 11 期 第 18 卷

黄连解毒汤含药血清对脂多糖诱导的肺泡上皮A549细胞炎症反应的作用研究

Study on the effect of Huanglian Jiedu decoction containing serum on lipopolysaccharide-induced inflammatory response in alveolar epithelial A549 cells

作者：刘心伟1，2胡玥1刘玉春1许晓娜3郭梦雨3任彦颖3刘莹3费冰3王春霞1，3李永伟1，2，3

英文作者：Liu Xinwei12 Hu Yue1 Liu Yuchun1 Xu Xiaona3 Guo Mengyu3 Ren Yanying3 Liu Ying3 Fei Bing3 Wang Chunxia13 Li Yongwei123

单位：1河南省中医院（河南中医药大学第二附属医院）检验科，郑州450002；2河南省防治耐药菌感染中医药重点实验室，郑州450002；3河南中医药大学第二临床医学院，郑州450046

英文单位：1Department of Clinical Laboratory Henan Province Hospital of Traditional Chinese Medicine (The Second Affiliated Hospital of Henan University of Chinese Medicine) Zhengzhou 450002 China; 2The Key Laboratory of Henan Province of Resistant Pathogen Prevention and Therapy by Traditional Chinese Medicine Zhengzhou 450002 China; 3The Second Clinical Medical College Henan University of Chinese Medicine Zhengzhou 450046 China

关键词：黄连解毒汤；肺泡上皮A549细胞；CD147；核因子κB；炎症反应

英文关键词：HuanglianJiedudecoction;AlveolarepithelialA549cells;CD147;Nuclearfactor-κB;Inflammatoryresponse

• 摘要：

• 目的  探讨黄连解毒汤对脂多糖诱导的肺泡上皮A549细胞炎症反应的作用，为临床合理应用该方提供理论依据和数据支撑。方法  将处于对数生长期的肺泡上皮A549细胞完全随机分为对照组、模型组和低、中、高剂量中药组。对照组加入5%正常大鼠血清；模型组加入含20 mg/L脂多糖的5%正常大鼠血清；低、中、高剂量中药组分别加入含20 mg/L脂多糖的5%、10%、15%黄连解毒汤含药血清。培养24 h后，噻唑蓝法检测肺泡上皮A549细胞活性，酶联免疫吸附试验法检测各组细胞上清中炎症因子白细胞介素1β（IL-1β）、IL-6和肿瘤坏死因子α含量，实时荧光定量聚合酶链反应和蛋白质印迹法分别检测各组细胞CD147和核因子κB p65的mRNA和蛋白表达。结果  模型组细胞上清中IL-1β、IL-6和肿瘤坏死因子α水平及细胞CD147和核因子κB p65的mRNA和蛋白表达水平均明显高于对照组（均P<0.01）。中剂量中药组和高剂量中药组IL-1β、IL-6和肿瘤坏死因子α水平均明显低于模型组和低剂量中药组［（133±14）、（119±14）ng/L比（219±20）、（189±20）ng/L，（308±31）、（288±28）ng/L比（400±39）、（367±35）ng/L，（362±37）、（310±32）ng/L比（473±46）、（448±46）ng/L］（P<0.01或P<0.05）。中剂量中药组和高剂量中药组CD147和核因子κB的mRNA和蛋白表达水平均明显低于模型组和低剂量中药组（P<0.01或P<0.05）。结论  黄连解毒汤可抑制脂多糖诱导的肺泡上皮A549细胞炎症因子表达，其作用机制与CD147和核因子κB表达下调有关。

• Objective  To explore the effect of Huanglian Jiedu decoction on lipopolysaccharide-induced inflammatory response of alveolar epithelial A549 cells, so as to provide theoretical basis and data support for the rational clinical application of this prescription. Methods  Alveolar epithelial A549 cells in the logarithmic growth phase were completely random divided into control group, model group, low, medium, and high-dose traditional Chinese medicine groups. The control group was given 5% normal rat serum; the model group was given 5% normal rat serum containing 20 mg/L lipopolysaccharide; the low, medium, and high dose traditional Chinese medicine groups were given 5%, 10%, and 15% Huanglian Jiedu decoction containing serum containing 20 mg/L lipopolysaccharide, respectively. After 24 h of incubation, the vitality of alveolar epithelial A549 cells was detected by the methyl thiazolyl tetrazoliun method. The levels of serum inflammatory cytokines interleukin 1β(IL-1β), IL-6 and tumor necrosis factor-α of alveolar epithelial A549 cells in each group were detected by enzyme linked immunosorbent assay, and the mRNA and protein expressions of CD147 and nuclear factor-κB p65 were detected by quantitative real-time-polymerase chain reaction assay and western blotting respectively. Results  IL-1β, IL-6 and tumor necrosis factor-α levels in cell supernatant, the mRNA and protein expression levels of CD147 and nuclear factors-κB p65 of cellular of model group were significantly higher than those of the control group(all P<0.01). The levels of IL-1β, IL-6 and tumor necrosis factor-α of medium-dose traditional Chinese medicine group and high-dose traditional Chinese medicine group were significantly lower than those of the model group and the low-dose traditional Chinese medicine group［（133±14）,（119±14）ng/L vs （219±20）,（189±20）ng/L;（308±31）,（288±28）ng/L vs （400±39）,（367±35）ng/L;（362±37）,（310±32）ng/L vs （473±46）,（448±46）ng/L］(P<0.01 or P<0.05). The mRNA and protein expression levels of CD147 and nuclear factor-κB of the medium- and high-dose traditional Chinese medicine groups were significantly lower than those of the model group and low-dose traditional Chinese medicine group(P<0.01 or P<0.05). Conclusion  Huanglian Jiedu decoction can inhibit the high expression of alveolar epithelial A549 cells inflammatory factors induced by lipopolysaccharide, which is associated with the down-regulation of CD147 and nuclear factor-κB.