二氧化硅对人肺泡Ⅱ型上皮细胞纤溶酶原激活物抑制因子-1和激活蛋白-1表达的影响  

Effects of silica on the expression of plasminogen activator inhibitor-1 and activator protein-1 in human alveolar epithelial cells type Ⅱ

作　　者：林志[1] 胡永斌[2] 冯德云[2] 初令[2] 王金胜[2] 曾庆富[2] 

机构地区：[1]中国药品生物制品检定所国家药品安全评价监测中心病理室,北京100050 [2]中南大学湘雅医学院病理教研室

出　　处：《中华劳动卫生职业病杂志》2005年第5期355-358,共4页Chinese Journal of Industrial Hygiene and Occupational Diseases

基　　金：国家自然科学基金资助项目(30170399)

摘　　要：目的研究SiO2对人肺泡Ⅱ型上皮细胞(A549)中纤溶酶原激活物抑制因子1(PAI1)及激活蛋白1(AP1)表达的影响,探讨SiO2致肺纤维化的发病机制。方法按SiO2(200μg/ml)与A549细胞共育时间的不同,实验分SiO2刺激0、4、8、16、24h组。利用逆转录聚合酶链反应(RTPCR)、Westernblot及免疫细胞化学SP法检测细胞PAI1mRNA和蛋白的表达;AP1(cjun/cfos)总蛋白及核蛋白表达的检测采用Westernblotting。结果SiO2刺激后,实验组PAI1mRNA及蛋白表达强度随时间延长依次增强,呈直线正相关(rmRNA=0.911,r蛋白=0.902,P<0.05),且实验组PAI1蛋白表达均高于对照组(0.36±0.03),其中24h组(0.73±0.01)表达最强;cjun和cfos核蛋白的表达亦明显高于对照组(0.52±0.02、0.15±0.01),cjun核蛋白的表达4h组(1.54±0.02)最强;cfos核蛋白表达8h组(0.36±0.01)最强;cjun和cfos核蛋白的表达均从孵育16h开始逐渐下降。AP1(cjun/cfos)总蛋白的表达各组间差异无统计学意义(P>0.05);PAI1阳性信号定位于胞浆和胞核。结论SiO2能够诱导A549细胞PAI1的表达,呈现出明显的时间-效应关系,核转录因子AP1在SiO2刺激的早期被激活。Objective To investigate the mechanism of silicosis by observing the effects of silica on the expression of plasminogen activator inhibitor-1 （ PAI-1 ） and activator protein-1 （ AP-1 ） in human alveolar epithelial ceUs type 11（A549）. Methods A549 cell and SiO2（200μg/ml） wereco-cultured for 0,4,8,16 and24 hrespectively.The reverse transcriptase polymerase chain reaction （RT-PCR）, Western blotting and SP immunocytochemistry were used for detections of the PAI-1 mRNA and protein expression.The nucleopmtein and total protein expression of AP-1 were investigated by Western blotting. Results The expression levels of PAI-1 mRNA and protein were increased in a time-dependent manner（ rmRNA = 0.911, rprotein = 0. 902, P 〈 0.05 ）. The expressions of PAI-1 mRNA and protein in experimental groups were higher than that in control group（ P 〈 0.05）and was the highest in 24 h group[ （0.73 ±0.01 ）vs（0.36 ± .03） ] .The nucleopmtein expressions of c-jun/c-fos in experimen- tal groups were also higher than those in control group（ P 〈 0.05）,and the nucleopmtein expression level of c-jun was the highest in 4 h group[ （ 1.54 ± 0.02）vs（0.56 ± 0.03） ] ;the nucleopmtein expression level of c-fos was the highest in 8 h group[ （0.36 ± 0.01 ）vs（0.15 ± 0.01 ） ]. Both c-jun and c-fos expression were decreased after 16 h, but the total protein expressionof c-jnn/c-fos had no difference in all experimental groups. The positive signal of PAI-1 was located in cytoplasm and nucleus. Conclusion SiO2 could induce PAI-1 expression of A549 in a timedependent manner, and AP-1 activation can be observed in early time.

关 键 词：二氧化硅 纤溶酶原激活物抑制因子-1 纤溶酶原激活剂 上皮细胞 纤溶酶原激活物抑制因子-1 肺泡Ⅱ型上皮细胞 激活蛋白-1 WESTERNBLOT 二氧化硅 逆转录聚合酶链反应 免疫细胞化学SP法 核蛋白表达 PAI-1 

分 类 号：R135.2[医药卫生—劳动卫生]