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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
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摘要 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.
作者 邹威凤 盛青 周晓婷 黎希 刘锐强 Zou Weifeng, Sheng Qing, Zhou Xiaoting, Li Xi, Liu Ruiqiang(Department of Respiratory Medicine, Guangzhou Chest Hospital, Guangzhou 510000, China)
出处 《国际呼吸杂志》 2018年第14期1057-1060,共4页 International Journal of Respiration
基金 广东省自然基金项目(2015A030310378、2016A030313422) 广州市卫生和计划生育科技项目(20161A011034)
关键词 人气道上皮细胞株 尼古丁 糖原合成酶激酶3β/β连环素 促纤维因子 Human bronchial epithelial cells Nicotine OSK3β/β-catenin Fibrogenic mediators
作者简介 通信作者:邹威凤,Email:szwei555@163.com
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