中文摘要：

目的：观察茶黄素对人中性粒细胞弹性蛋白酶（human neutrophil elastase，HNE）诱导气道黏液高分泌的作用。方法：采用HNE刺激人肺腺癌细胞A549，导致黏液高分泌，以茶黄素双没食子酸酯（theaflavin-3，3＇-digallate，TF3）单体进行干预。首先用四甲基偶氮唑盐光吸收法（MTT）测定细胞活性，确定作用剂量，其后细胞分为4组进行实验，分别为A组：对照组；B组：HNE（50nmol／L）刺激24h；C组：用TF3（浓度分别为50、100、200μg/ml）作用24h后，再予HNE刺激24h；D组：先用EGFR阻断剂AG1478（5μmol／L）处理30min后，也予HNE刺激24h。并测定不同剂量和TF3（200μg/ml）不同作用时间（12h、24h、36h）的黏蛋白变化。RT-PCR检测备组黏蛋白（MUC）5ACmRNA的变化；酶联免疫吸附测定法（ELISA）观察MUC5AC蛋白表达的变化。结果：HNE能导致A549的MUC5ACmRNA和MUC5AC蛋白显著增高，而给予TF3预处理后，与HNE刺激组比，两指标均明显下调，且呈浓度剂量和作用时间正相关，其差异有统计学意义；茶黄索纽与AG1478组比较，后者抑制作用更为明显；与对照纽比，仍有增高。结论：荼黄素双没食子酸酯（TF3）单体可显著下调炎性气道的黏液高分泌，但并不能完全抑制炎性黏液的分泌。

英文摘要：

Objective： To investigate the effects of theaflavin-3,3＇-digallate （TF3） on airway mucous hypersecretion induced by human neutrophil elastase（HNE）. Methods： Human lung adenocarcinoma cells-AS49 mucous hypersecretion was induced by the stimulation of HNE. The cell activity of TF-3 treated was assessed by MTT method. In this study, we divided into 4 groups： a negative control group（ A）, HNE treatment group（ B）, TF3 pre-treatment group（C） and AG1478 pre-treatment group（D）. Group B was treated with HNE （50nmol/L） for 24 hours. Group C were pre-treated with TF3 （50,100,200 μg/ml） for 24 hours, group D was pre-treated with AG1478（5 μmol/L）for 30 minutes respectively, then they were stimulated with HNE （50 nmol/L） for 24 hours. We tested the change of the mucin at 12 h, 24 h and 36h. The reverse transcriptase-polymerase chain reaction （RT-PCR） was used to examine a changes of MUC5ACmRNA. The protein expression changes of MUCSAC was tested by Enzyme-hnked immunosorbent assay （ELISA）. Results： The MUCSACmRNA expression and protein expression of the B group were 0.86 ±0.03 and 163.22 ±7.34 respectively, both significantly higher than those of the A group（ 0.22 ±0.02 and 102. 28±3.87 both P 〈0.01 ）. The above two indexes were down regulated obviously after the cells were pretreated by TF-3 compare with HNE group. The difference was significant statistically （ P 〈 0.01 ） and it presented positive correlation of the concentration and the action time. On the other hand, we also observed that the inhibitory action of D group were stronger than C group. However, there were significantly augmented between C group and A group （ P 〈 0.01 ）. Conclusion： We conclude that TF3 were able to inhibit airway mucous hypersecretion induced by inflammatory factor obviously. But it could not completely inhibited mucous hypersecretion.