【摘要】 目的观察糖原合成酶激酶3β(glycogen synthase kinase 3β,GSK3β)在气道上皮细胞(airway epithelialcells,AECs)损伤修复中的变化。方法利用吸烟/香烟提取物(cigarette smoke extract,CSE)、脂多糖(lipopolysacchor-id,LPS)和博来霉素(bleomycin,BLM)刺激分别建立几种体内、外气道上皮损伤模型,采用免疫细胞荧光、共聚焦成像和Western blot方法,观察GSK3β在几种不同损伤模型中的表达变化。结果①荧光共聚焦成像:GS3β在对照组小鼠AECs胞质内呈均匀分布的红色荧光颗粒,而吸烟、LPS和BLM三种体内模型中AEC胞质内红色荧光均明显减弱;免疫细胞荧光:GSK3β在对照组AECs胞质内呈强阳性均匀分布的红色荧光颗粒,而CSE、LPS、BLM刺激后的三种体外模型GSK3β在胞质内红色荧光均明显减弱。②Western blot:在体内模型中,吸烟组、LPS刺激组及BLM刺激组GSK3β表达均低于对照组,其中吸烟1w、LPS刺激1d、BLM刺激7d时表达最低(P≤0.05);而P-GSK3β均高于对照组,其中吸烟1w、LPS刺激7d和14d达到高峰(P<0.05)。在体外模型中,以上三种刺激均可导致抑制性磷酸化GSK3β的水平升高并呈浓度依赖性(P<0.05),而GSK3β表达则完全相反,随着浓度的升高趋势越明显(P<0.05)。结论吸烟/CSE,LPS、BLM刺激致AEC损伤修复过程中伴有GSK3β表达的活性改变,提示其在损伤修复中发挥重要作用。更多还原

【Abstract】 Objective To observe the changes in the expression of glycogen synthase kinase 3β(GSK3 β in the injured and repaired airway epithelial cells(AECs).Methods We established several in vitro and in vivo models of the injured and repaired airway epithelial cells induced by smoking,cigarette smoke extract(CSE),lipopolysaccharide(LPS) or bleomycin(BLM).The localizations and levels of GSK3β expression were observed by immunofluorescence or Western blot respectively.Results Immunofluorescence showed that GSK3 β were both uniformly expressed in plasma of AECs of the control groups and the experimental groups.There were high levels of GSK3β in the control groups,whereas the levels of GSK3β were decreased in the experimental groups.Western blot showed that the levels of GSK3β in AECs of the three in vivo models obviously decreased(P<0.05 vs the control groups respectively);the levels of P-GSK3β in the experimental groups were higher than those in the corresponding control groups(P<0.05). Moreover,it was found that in vitro cigarette smoke extract(CSE),lipopolysaccharide(LPS) or bleomycin(BLM) notably inhibited GSK3β by increasing inactive phosphorylated GSK3β(P<0.05 vs the control groups respectively) and reducing GSK3 β expression(P<0.05 vs the control groups respectively).Conclusion The expression and the activity of GSK3β are changed in several in vitro and in vivo models of the injured and repaired AECs, suggesting that GSK3β may play an important role in the process of repair of airway epithelial injury.更多还原