中文摘要：

目的探讨丙烯醛调控的糖皮质激素受体（GRs）类小泛素（SUMO）化水平对气道黏液高分泌的影响。方法构建重组真核表达载体质粒pcDNA3．1．EGFP．flag．SUM01，酶切电泳鉴定。常规培养16HBE细胞，随机分为9组：A：空白对照组；B：丙烯醛刺激组；C：地塞米松组；D：丙烯醛刺激＋地塞米松组；E：SUMO相关修饰物1（SUM01）转染组；F：空质粒转染组；G：SUM01转染＋丙烯醛刺激组；H：SUM01转染＋丙烯醛刺激＋地塞米松组；I：空质粒转染＋丙烯醛刺激＋地塞米松组。每个实验组设5复孔，重复4次实验用于统计学分析。荧光显微镜下观察转染成功率，通过免疫共沉淀以及Western印迹法分析各实验组GRsSUMO化水平的变化，反转录-PCR法比较各实验组中黏蛋白（MUC）5AC转录水平的差异。用酶联免疫吸附法测定细胞培养上清液中MUC5AC的分泌量。结果B组GRsSUMO化水平（0．079±0．023）低于A组（0．446±0．068）（t=-23．13，P=0．000）；D组GRsSUMO化水平（0．574±0．018）低于c组（0．843±0．028）（t=-36．34，P=0．000）。H组的MUC5ACmRNA转录水平（0．330±0．063）低于D组（0．617±0．190）（q=-7．80，P=0．000）。细胞培养上清液中MUC5AC分泌水平显示，G组与B组差异无统计学意义，而H组[（0．416±0．092）μg／L）]显著低于D组[（0．663±0．104）μg／L]和G组[（0．740±0．343）μg／L]（q=-7．31，-9．59；P=0．001，0．000）。结论丙烯醛可下调GRsSUMO化水平，并降低地塞米松对MUC5AC转录的抑制作用。

英文摘要：

Objective To explore the variation of small ubiquitin-related modification （SUMO） level of glucocorticoid receptors （ GRs ） exposed to acrolein stimulation as well as the influence of the variation on mucus hypersecretion. Methods The recombinant plasmid was constructed by inserting small ubiquitin-like modifier 1 （SUMO1） cDNA into eukaryotic expression plasmid pcDNA3.1-EGFP with a flag sequence marker. The recombinant plasmid peDNA3.1-EGFP-flag-SUMO1 was identified by enzyme digestion analysis. The 16HBE cells were cultured and randomized divided into 9 following groups： A. control; B. aerolein stimulated; C. dexamethasone incubated; D. acrolein stimulated and dexamethasone incubated; E. SUMO1 transfected; F. pcDNA3.1-EGFP vector transfected; G. SUMO1 transfeeted and acrolein stimulated; H. SUMO1 transfected, acrolein stimulated and dexamethasone incubated; I. peDNA3.1-EGFP vector transfected, acrolein stimulated and dexamethasone incubated. Each group consisted of 5 parallel wells and experiments were repeated for 4 times for statistical analysis. The transfection efficiency was evaluated by the expression of enhanced green fluorescent protein （EGFP） via fluorescence microscope. The SUMO modification level of GRs was measured with co-immunoprecipitation and Western blot. The transcription level of mucin （MUC） 5AC was evaluated with reverse transcription- polymerase chain reaction （RT-PCR）. The MUC5AC secreted in supernatant was measured by enzyme- linked immunosorbent assay （ELISA）. Results The SUMO modification level was much lower in group B （0. 079 ±0. 023） than that in group A （0. 446 ±0. 068） （t = -23.13, P =0. 000） and in group D （0. 574 ± 0. 018） than that in group C （ 0. 843 ± 0. 028 ） （ t = - 36. 34, P = 0. 000 ）. The transcription level of MUCSAC was significantly lower in group H （ 0. 330 ± 0. 063 ） than group D （ 0. 617 ± 0. 190 ） （ q = -7. 80, P = 0. 000 ）. Through ELISA, there was no significance between groups G and B in the