博来霉素致肺纤维化大鼠不同时间点吸及STAT1反义寡核苷酸的疗效比较  

Therapeutic effects of aerosolized signal transducer and activator of transcription 1 antisense oligonucleotide administered at different time points on bleomycin-induced pulmonary fibrosis： experiment with rats

作　　者：李晶[1] 曾鸣[1] 朱晨[1] 王文军[1] 湛晓勤[1] 范贤明[1] 

机构地区：[1]四川省泸州医学院附属医院呼吸内科,646000

出　　处：《中华医学杂志》2009年第12期831-835,共5页National Medical Journal of China

基　　金：国家自然科学基金（30570814）;四川省教育厅支持计划（05011）

摘　　要：目的探讨雾化吸入信号转导和转录活化因子1（STAT1）反义寡核苷酸（ASON）干预肺纤维化的最佳给药时机。方法Wistar雌性大鼠25只随机均分为博来霉素（BLM）组、ASON0d组、ASON7d组、ASON14d组和生理盐水（NS）组，前4组气管内灌注BLM建立肺纤维化模型，NS组气管内灌注NS。ASON0、7、14d组分别于气管内灌注BLM后立即、第7和14天开始雾化吸入STAT1 ASON；NS组和BLM组雾化吸入NS。气管内灌注BLM后第28天处死各组大鼠，取肺组织分别行HE和Masson染色，观察肺泡炎和纤维化情况并评分；酶联免疫吸附试验（ELISA）测定支气管肺泡灌洗液（BALF）中转化生长因子β（TGF-β）和肿瘤坏死因子仅（TNF-α）浓度。结果肺组织病理学观察显示ASON0d组大鼠肺泡炎和肺纤维化程度明显轻于BLM组和ASON14d组，肺泡炎评分（1．80±0．84）和肺纤维化评分（2．60±0．55）均明显低于BLM组（2．40±0．55、4．40±0．55）、ASON7d组（2．20±0．45、3．00±0．71）和ASON14d组（2．20±0．84、4．00±1．00）（均P〈0．05）；ASON7d组肺纤维化评分也明显低于BLM组和ASON14d组（均P〈0．05）。ASON0d组BALF中TGF—β与TNF-α浓度[（48．11±3．46）pg／ml、（1．93±0．14）ng／ml]均明显低于BLM组[（57．67±2．46）pg／ml、（2．45±0．25）ng／ml，均P〈0．05]，TGF—β浓度明显低于ASON7d组[（51．42±3．57）pg／ml]和ASON14d组[（55．83±1．79）pg／ml]（均P〈0.05）；ASON7d组BALF中TGF—β浓度也明显低于BLM组和ASON14d组（均P〈0．05）。结论早期雾化吸入STAT1ASON对BLM致肺纤维化大鼠的肺纤维化形成有明显阻抑作用，用药越早效果越好，提示雾化吸入STAT1 ASON有可能成为肺纤维化的早期干预手段。Objective To investigate the curative effects of inhaling signal transducer and activator of transcription 1 （STAT1） antisense oligonucleotidc （ASON） on alveolitis and pulmonary fibrosis and the best administration time. Methods Twenty-five adult female Wistar rats were randomly divided into 5 equal groups ： BLM group, undergoing intra-tracheal perfusion of BLM so as to establish animal models of alveolitis and pulmonary fibrosis and then inhaling aerosolized normal saline （NS）; NS group undergoing intratracheal perfusion of NS and then inhaling aerosolized NS; ASON 0 d group, undergoing intra-tracheal perfusion of BLM and then inhaling aerosolized STAT1 ASON 3 ml immediately; ASON 7 d group, undergoing intra-tracheal perfusion of BLM and then inhaling STAT1 ASON 3 ml 7 days later; and ASON 14 d group undergoing intra-tracheal perfusion of BLM and then inhaling aerosolized STAT1 ASON 3 ml 14 days later. Aerosolized inhalation was repeated once every other day for 4 times. Twenty-eight days after intratracheal perfusion the rats were sacrificed with their lungs taken out to undergo pathological examination. NS was infused into the right lungs to get bronchoalveolar lavage fluid （BALF）. ELISA was used to examine the concentrations of transforming growth factor-β （TGF-β） and tumor necrosis factor-α（TNF-α） in the BALF. Results The pathology result of the lung tissues showed that compared with the BLM and ASON 14 d groups, the atveolitis and pulmonary fibrosis of the ASON 0 d group were obviously milder. The scores of alveolitis and pulmonary fibrosis of the ASON 0 d group were （1. 80 ± 0. 84 ） and （2. 60 ±0. 55 ） respectively, both significantly lower than those of the BLM group [ （ 2.40 ± 0. 55 ） and （4.40 ± 0. 55 ） respectively ] and those of the ASON 7 d group [ （2.20 ± 0.45 ） and （ 3.00 ± 0.71 ） respectively ] （ all P 〈 0. 05）. The scores of pulmonary fibrosis of the ASON 7 d group was significantly lower than those of the BLM and ASON 14 d groups

关 键 词：STAT1转录因子类 寡核苷酸类 反义 肺纤维化 大鼠 Wistar 

分 类 号：R285.5[医药卫生—中药学] R735.2[医药卫生—中医学]