中文摘要：

目的观察BTB-cap′n′collar（CNC）同源异构体1（Bachl）-siRNA腺病毒载体对TGF-β1诱导后的小鼠肺成纤维细胞（MLF）中抗氧化因子及纤维化相关细胞因子表达的影响。方法构建Bachl siRNA腺病毒载体及空病毒载体，5ng/ml TGF-β1处理MLF24h，分别感染空病毒载体及Bachl siRNA腺病毒载体。RT—PCR及蛋白印迹法分别检测细胞中Bach1、血红素加氧酶1（HO-1）、谷胱甘肽过氧化酶1（GPxl）及还原型烟酰胺腺嘌呤二核苷酸磷酸氧化还原酶1（NQ01）的mRNA和蛋白表达水平；ELISA检测细胞上清液中纤维化相关因子TGF-β1及IL-6的水平。多个组间的比较采用单因素方差分析，两两比较采用LSD检验。结果TGF-β1诱导后MLF中Bachl mRNA水平（2．127±0．089）和蛋白表达较空白对照组mRNA（1．000±0．067,t=-21．77，P〈0．01）及蛋白表达明显升高，细胞上清中纤维化相关因子TGF-β1（52±6）及IL-6（34±6）较空白对照组（26±4，t=-11．11，P〈0．01及20±5，t=-5．32，P〈0．01）的表达也明显升高；但抗氧化因子HO-1及GPx1的mRNA水平（0．403±0．040及0．567±0．112）和蛋白表达较对照组mRNA（1．000±0．181，t=25．57，P〈0．01及1．000±0．212，t=6．68，P〈0．05）和蛋白表达明显减低；BachlsiRNA处理后，MLF中Bachl mRNA水平（0．153±0．015）和蛋白表达较TGF-β1组及空载体组的mRNA水平（2．129±0．089及1．973±0．035，F=1835．95，P〈0．01）和蛋白表达明显降低；细胞上清中TGF-β1（26±3）和IL-6（11±3）的表达水平也较TGF-β1组（52±6及34±6）及空载体组（49±5及33±6）明显降低（F=22．25，P〈0．01及F=28．38，P〈0．01），但HO-1（3．303±0．294）和GPx1（1．840±0．231）mRNA水平和蛋白表达较TGF-β1组（0．403±0．040及0．567±0．112）及空载体组（0．353±0．057及0．667±0．090）明显升高（F=53．90，P〈0．01及F=526．25，P〈0．01）。结论沉默Bach1能?

英文摘要：

Objective To investigate the effects of adenovirus vectors for BTB and capOn＇collar protein homology 1 （Bachl） small interfering RNA （siRNA） on antioxidanffactors and fibrosis related cytokines in lung fibroblasts （MLF） in transforming growth factor （TGF）-β1 induced mouse. Methods Bachl siRNA recombinant adenovirus vectors and blankadenovirus vectorwere constructed, then the MLF cells were incubated with TGF-β1 （5 ng/ml） for 24 h and infected with blankvector and successful constructed Bachl siRNAs. The messenger RNA （mRNA） and protein expression of Bachl, heine oxygenase （HO）-I, glutathione peroxidase 1 （GPxl） and nicotinamide-adenine dinucleotide phosphate： quinone oxidoreductase-1 （NQO1） in cell super- natants were determined by reverse transcription-polymerase chain reaction （RT-PCR） and Westein blot. Changes of fibrosis-related cytokines including TGF-β1 and interleukin （IL）-6 levels in cell supernatants were determined by enzyme-linked immunosorbent assay （ELISA）. One-way analysis of variance （ANOVA） was performed for multiple group comparisons and LSD test was used to compare the two groups. Results Bachl mRNA （2.127±0.089） and protein expression increased significantly after TGF-β1 stimulation compared with blank group （1.000±0.067, t=-21.77, P〈0.01）, as well as the expression of fibrosis-related cytokines TGF-β1 （52±6） and IL-6 （34±6） in cell supernatants increased significantly after TGF-β1 stimulation compared with blank group （26±4, t=-11.11, P〈0.01 and 20±5, t=-5.32, P〈0.01）, but the mRNA expression of HO-1 and GPxl （0.403±0.040 and 0.567±0.112） also the protein expression decreased significantly compared with mRNA （1.000±0.181, t=25.57, P〈0.01 and 1.000±0.212, t=6.68, P〈0.05） and protein expression in blank group. Follow the Bachl siRNA treatment, Bachl mRNA （0.153±0.015） and protein levels were significantly downregulated compared with mRNA （2.129±0.089 and 1.973±0.035, F=1835.95,