中文摘要：

目的探讨细胞因子信号转导抑制因子1（SOCS1）对脂多糖诱导的气道黏液高分泌的抑制作用及其机制。方法培养人支气管上皮16HBE细胞，根据脂多糖刺激时长将细胞分为0、0．5、1、6、12及24h组，确定后续实验作用时长。根据不同的转染或处理措施，将细胞分为七组，分别为未转染组、空质粒组、野生型SOCS1组、阴性siRNA组、SOCSl1小干扰RNA（SOCS1-siRNA）组、生理缓冲液组和Filgotinib组，再给予脂多糖刺激，以同体积磷酸盐缓冲液处理细胞为对照。酶联免疫吸附（ELISA）法检测以上各组细胞裂解液中黏蛋白5AC（MUC5AC）（txg）相对裂解液总蛋白（mg）的表达量。Western印迹法分别检测细胞裂解液中SOCS1、磷酸化JAK1和STAT1表达量，分别以β-肌动蛋白或总JAKI和STAT1蛋白表达量为内参。结果脂多糖可诱导MUC5AC蛋白高表达，0、0．5、16、12、24h组MUC5AC的相对表达量分别为（2．86±0．20）、（3．42±0．29）、（3．43±0．12）、（10．22±0．96）、（14．56±1．12）、（14．15±1．34）μg／mg，而SOCSl蛋白则呈反向表达；6h组MUC5AC蛋白和SOCS1蛋白均处于中间水平，故选择6h为脂多糖刺激时长。野生型SOCS1组细胞内SOCS1呈过表达状态，并且同Filgotinib组一样，JAK1和STAT1磷酸化水平均显著降低；野生型SOCS1组和Filgotinib组MUC5AC的相对表达量均显著低于未转染组[（4．04±0．65）、（7．02±0．83）比（10．37±1．00）μg／mg]（均P〈0．05）。反之，SOCS1-siRNA组细胞内SOCS1表达明显下降，JAK1和STAT1磷酸化水平增强；SOCS1-siRNA组MUC5AC的相对表达量显著高于阴性siRNA组[（13．69±1．32）比（11．01±1．41）μg／mg]（P〈0．05）。结论SOCS1可通过对JAK1／STAT1信号通路的负调节作用，抑制脂多糖诱导的MUC5AC高表达。

英文摘要：

Objective To explore the inhibitory role of suppressor of cytokine signaling 1 （ SOCS1 ） in lipopolysaceharide （LPS）-induced mucin5AC （MUC5AC） hypersecretion and the potential mechanism involved in this process. Methods The human bronchial epithelial cells 16HBE were divided into 0, 0. 5, 1, 6, 12 and 24 h groups according to the time of LPS challenge. In gain- and loss- of functions experiments, wild-type SOCSI and SOCS1 -targeted siRNA （SOCSI-siRNA） were synthesized to identify the function of SOCS1 in LPS-mediated MUC5AC hypersecretion, and named wild-type SOCS1 group and SOCSI-siRNA group, respectively, and the non-transfected group and non-targeted siRNA group were used as controls. In Filgotinib group, the specific inhibitor of Janus kinase 1 （ JAK1 ） , Filgotinib, was used to detect the role of JAK1/signal transducer and activator of transcription 1 （ STAT1 ） signaling pathway in LPS challenge, and the aqueous physiological buffer group was used as the control. The production of MUC5AC protein was measured by enzyme linked immunosorbent assay （ELISA） , and the amount of MUC5AC proteinwas normalized to the total protein in cell lysates and was expressed as μg/mg cell lysates. The proteins expressions of SOCS1, phosphorylation of JAK1 and STAT1 were measured by Western blot, and the total expression of its protein （for JAK1 and STAT1 ） or β-actin （ for SOCS1 ） was used as the loading control. Results Compared to 0 h group, LPS induced a robust induction in MUC5AC expression, the expression levels of MUC5AC in 0, 0. 5, 1, 6, 12 and 24 h groups were （2. 86 ±0. 20）, （3.42 ±0. 29）, （3.43 ± 0. 12）, （ 10. 22 ± 0.96）, （ 14. 56 ± 1.12）, （ 14. 15 ±1.34）μg/mg, in association with a decrease of SOCS1 expression. And in 6 h group, the expressions of MUC5AC and SOCS1 were both medium up-regulated （ all P 〈 0. 05 ）. Consequently, the application of LPS for 6 h was selected as the optimal responses period in the ensuing experiments. Compared to the