葛根素对火药烟雾诱导支气管上皮细胞凋亡的防护作用

Puerarin protects human bronchial epithelial cells from apoptosis induced by gunpowder smog

目的探讨葛根素对火药烟雾诱导的支气管上皮细胞(BEAS-2B)凋亡的保护作用及其机制。方法体外培养BEAS-2B细胞,随机分为对照组(不加任何干预)、烟熏组(火药烟雾4g,作用10min)、烟熏+葛根素组(12.5、25、50、100μg/ml)。细胞接种12h后加入葛根素,继续培养至24h后烟雾作用10min,继续培养2h后进行检测。采用CCK-8试剂盒检测细胞活力,Hoechst染色观察细胞凋亡情况,流式细胞仪检测Annexin V-PI双染细胞及caspase-3阳性率。结果与对照组相比,烟熏组BEAS-2B细胞活力明显下降(P<0.01),不同浓度葛根素能够不同程度地对抗烟熏的作用,其最佳保护浓度为25μg/ml。与烟熏组相比,烟熏+葛根素组Hoechst染色细胞凋亡率、Annexin V-PI双染细胞凋亡率及caspase-3阳性率均明显降低(P<0.05,P<0.01)。结论火药烟雾可致体外BEAS-2B细胞凋亡,而葛根素对火药烟雾诱导的BEAS-2B气管上皮细胞凋亡具有一定防护作用。

Objective To investigate protective effects of puerarin on the human bronchial epithelial（BEAS-2B） cell line against apoptosis caused by gunpowder smog and its mechanisms. Methods BEAS-2B cells cultured in vitro were randomly divided into control group, smog group（the group treated with 4g gunpowder smog for 10min）, and smog ＋ puerarin group [puerarin group, the cells were pre-incubated with various concentrations of puerarin（12.5, 25.0, 50.0, 100.0μg/ml） and then exposed to smoke]. Puerarin was added into the cells after innoculation for 12 h and then the cells were sequentially cultured for 24 h and followed by exposure to smoke for 10 min. After being cultured again for 2h, the smoked cells were examined for cell viability using Cell Counting Kit-8（CCK-8）, cell apoptosis was obser ved using Hoechst33258 nucleus staining, and positive rates of Annexin V-PI staining cells and caspase-3 were determined with flow cytometer. Resu lts Compared with control, treatment of BEAS-2B cells with 4g gunpowder smog induced a characteristic apoptotic cell death（P〈0.01）. Pretreatment with various concentrations of puerarin antagonized the action of gunpowder smog in different degrees. The 25μg/ml was determined as the optimal effective concentration of puerarin. Compared with smog group, the apoptosis rate of BEAS-2B cells and positive rates of Annexin V-PI staining cells and caspase-3 were decreased significantly in smog ＋ puerarin group（P〈0.05, P〈0.01）. Conclusion Gunpowder smog can induce apoptosis of BEAS-2B cells in vitro, while pretreatment with puerarin could protect BEAS-2B cells against apoptosis induced by gunpowder smog.