糖原合成酶激酶3参与尼古丁调控支气管上皮细胞促纤维因子的分泌

Nicotine induces the expression of fibrogenic mediators via glycogen synthase kinase 3 in human airway epithelial cells

目的本研究旨在探讨尼古丁是否通过糖原合成酶激酶3β（GSK3β）的活性变化及其对β-连环素信号的影响调控气道上皮细胞基质金属蛋白酶9（MMP-9）、血小板衍生生长因子AB（PDGF—AB）和PDGF-BB分泌。方法尼古丁（6×10μmol／L）刺激人气道上皮细胞株（16HBE），采用Western blotting方法检测刺激24h后细胞中GSK3β和抑制性磷酸化GSK3β（P—GSK3β）的表达水平。预先加入10μmol／LGSK3~特异性抑制剂SB216763孵育1h，再加入尼古丁刺激16HBE24h，细胞免疫荧光和Western blotting方法检测D-连环素胞浆和胞核的表达，用酶联免疫吸附试验检测MMP-9、PDGF-AB和PDGF-BB分泌。结果尼古丁刺激24h后P—GSK313的表达较未予尼古丁刺激的正常对照组明显升高，GSK3β的表达下降。加入SB216763后尼古丁刺激24h组，β-连环素胞浆和胞核的表达较尼古丁组明显增加，同时MMP-9、PDGF-AB和PDGF—BB分泌量显著增加。结论GSK3β／β-连环素信号通路参与了尼古丁调控支气管上皮细胞MMP-9、PDGF—AB和PDGF-BB分泌。

Objective The aim of the present study was to test whether nicotine induces the secretion of matrix metalloprotein-9 （MMP-9）, platelet-derived growth factor-AB （PDGF-AB） and PDGF-BB in human bronchial epithelial cells （HBECs） in vitro via the GSK3β/β-catenin signaling pathway. Methods Cultured HBECs were exposed to nicotine （6×10^-6 mol/L） for 24 h. The expression of GSK3β and P-GSK3β was assessed by Western blotting. HBECs were incubated with GSK3β inhibitor SB216763 for 1 h and subsequently stimulated with nicotine. The cytoplasmic and nuclear of β-catenin was assessed by Western blotting and immunofluorescence, the secretion of fibrogenic mediators was assessed bY enzyme linked immunosorbent assay. Results We showed that cells exposed to nicotine for 24 h exhibited significantly increased P-GSK3β expression and decreased GSK3β expression, SB216763 significantly increased cytoplasmic accumulation and nuclear translocation Of β-catenin and increased the secretion MMP-9, PDGF-AB and PDGF-BB from HBECs. Conclusions These results suggest that nicotine induces the secretion of MMP-9, PDOF-AB and PDGF-BB in HBECs in vitro via the OSK3β- catenin signaling pathway.