STAT1 ASON对肺纤维化大鼠BALF细胞因子和肺组织Ⅰ、Ⅲ型胶原mRNA表达的影响

Effect of signal transducer and activator of transcription 1 antisense oligodeoxynucleotides on inflammatory mediators,typeⅠand type Ⅲ collagen mRNA of rat pulmonary fibrosis

目的:观察信号转导子与转录活化子1(STAT1)反义寡核苷酸(ASON)雾化吸入对博莱霉素(BLM)致肺纤维化大鼠肺泡灌洗液(BALF)PDGF-BB(血小板衍生生长因子-BB)、TNF-α(肿瘤坏死因子-α)、TGF-β1(转化生长因子-β1)、IFN-γ(干扰素-γ)表达及肺组织Ⅰ、Ⅲ型胶原mRNA表达的影响。方法:取45只健康雌性Wistar大鼠随机分成ASON组、BLM组和生理盐水(NS)组各15只,气管内分别灌注BLM(ASON组、BLM组)和NS(NS组)复制肺纤维化模型和空白对照,ASON组于第0、2、4、6天雾化吸入STAT1ASON,BLM组和NS组雾化吸入NS,各组分别于第7、14、28天均处死动物5只。观察肺泡炎和肺纤维化改变;收集BALF测定PDGF-BB、TNF-α、IFN-γ、TGF-β1的水平;测定肺组织Ⅰ、Ⅲ型胶原mRNA表达。结果:ASON组第7、14、28天肺泡炎和肺纤维化程度明显低于同时间点BLM组(P<0.05)。BLM组各时间点BALF中PDGF-BB、TNF-α、TGF-β1表达水平较NS组升高(P<0.05),ASON组较BLM组各时间点均降低(P<0.05);BLM组BALF中IFN-γ表达水平各时间点均低于NS组(P<0.05),但ASON组各时间点均较BLM组高(P<0.05)。ASON组和BLM组第7、14、28天Ⅰ、Ⅲ型胶原mRNA表达高于NS组(P<0.05);ASON组第7、14、28天Ⅰ、Ⅲ型胶原mRNA的表达均较同时间点BLM组降低(P<0.05)。结论:STAT1ASON雾化吸入能够减轻BLM诱导的肺泡炎和肺纤维化程度,其机制可能与抑制BALF中TGF-β1、PDGF-BB、TNF-α的表达,抑制IFN-γ下降而减少肺组织胶原的沉积有关。

AIM： To investigate the effect of aerosolized signal transducer and activator of transcription 1 （ STAT1 ） antisense oligodeoxynucleotides （ASON） on the expression of inflammatory mediators in bronchoalveolar lavage fluid （BALF） and type Ⅰ and type Ⅲ collagen mRNA of the bleomyein-induced rat pulmonary fibrosis. METHODS： 45 adult female Wistar rats were randomly divided into 3 groups： normal saline （NS） group, bleomycin （BLM） group and ASON group. BLM group and ASON group were intratracheally instilled with bleomycin （BLM） while NS group was instilled with NS. NS group and BLM group were aerosolized with NS while ASON group was aerosolized with STAT1 ASON on day 0, 2, 4 and 6 after intratracheal administration. Then each group was divided into 3 subgroups and the rats were sacrificed on day 7, 14 and 28. The concentration of IFN-γ, TNF-α, TGF-β1 and PDGF-BB in BALF was detected. The lung tissues were removed and HE and Masson staining was performed to observe the extent of alveolitis and fibrosis. The mRNA levels of type Ⅰ and type Ⅲ collagen in the lung tissues were measured. RESULTS： Compared with BLM group, the scores of alveolitis and fibrosis in ASON group were remarkably meliorated （ P 〈 0.0.5）. Compared with NS group, the concentration of TNF-α, TGF-β1 and PDGF-BB in BALF in BLM group was significantly increased, but it was lower in ASON group than in BLMA group （P〈0. 05）. The concentration of IFN-γ in BALF was lower in BLM group than in NS group （P〈0.05）, but it was higher in ASON group than in BLM group （ P 〈 0.05）. The mRNA levels of type Ⅰ and type Ⅲ collagen at various time points in ASON group were significantly lower than those in BLM group （ P 〈 0. 05）. CONCLUSION： The aerosolized STAT1 ASON has anti-fibrosis effect, which may result from the lessened production of type Ⅰ and type Ⅲ collagen through reducing the concentration of cytokines in BALF such as TNF-α, PDGF-BB and TGF-β1 and inhibiting the decline of IFN-γ in BALF.