矽肺纤维化体外细胞模型的建立  

Development of a cellular model for studying silicotic fibrosis in vitro induced by silicon dioxide

作　　者：马海燕[1] 毛国根[2] 宋志芳[2] 

机构地区：[1]杭州医学高等专科学校,杭州310012 [2]浙江大学医学院劳动卫生与环境卫生研究所

出　　处：《中华劳动卫生职业病杂志》2001年第4期289-291,共3页Chinese Journal of Industrial Hygiene and Occupational Diseases

基　　金：国家自然科学基金资助项目 ( 39670 62 8)

摘　　要：目的　根据SiO2 对不同类型细胞的增殖效应 ,建立矽肺纤维化的体外细胞模型。方法　以不同剂量的SiO2 刺激SD大鼠肺泡巨噬细胞 (PAM)及佛波酯 (PMA)诱导分化THP 1(具有PAM特性的人血单核细胞株 )细胞的培养上清液 ,分别作用于中国仓鼠肺成纤维细胞 (CHL)和貂肺上皮细胞 (CCL 64) ,采用四唑盐 (MTT)比色法测定其增殖效应 (以A570nm值表示 )。结果　在 0、5 0、10 0、2 0 0、5 0 0 μg/mlSiO2 浓度范围内 ,用体积分数为 5 %的SiO2 刺激的PAM和体积分数为 10 %的SiO2 刺激的PMA primedTHP 1细胞上清液分别作用于CHL和CCL 64细胞 ,均可引起细胞增殖 ,2 0 0、5 0 0 μg/mlSiO2 剂量组与对照组的差异有显著性 ,并存在剂量 -效应关系 (CHL :r1=0 .914 ,P <0 .0 5 ;r2 =0 .980 ,P <0 .0 1。CCL 64:r1=0 .975 ,P <0 .0 1;r2 =0 .95 6,P <0 .0 1)。以体积分数为 10 %的SiO2 刺激PAM的上清液分别作用于CHL和CCL 64,5 0 μg/mlSiO2 组的A570nm值为 :CHL :0 .66± 0 .0 5 ,CCL 64:0 .2 1± 0 .0 4 ,与对照组 (CHL :0 .74±0 .0 4 ;CCL 64:0 .2 6± 0 .0 4 )的差异有显著性 (P <0 .0 5 ) ,均表现为抑制增殖作用。结论　利用体积分数为5 %的SiO2 刺激的PAM上清液和体积分数为 10 %的SiO2 刺激的PMA primedTHPObjective Based on the effects of silicon dioxide on the proliferation of different kinds of cells,a cellular model for studying pulmonary fibrosis induced by silicon dioxide was developed. Methods The pulmonary alveolar macrophage(PAM) or the phorbol 12-myristate 13-acetate(PMA)-primed THP-1 cell supernatants were prepared by incubating the cells in the serum-free RPMI 1640 with various concentrations of SiO 2 for 24 hours(PAM) or 36 hours(PMA-primed THP-1).CHL and CCL-64 cells were incubated with the prepared supernatants for 24 hours.After incubation,the proliferation of CHL and CCL-64 cells were measured using MTT assay. Results 5%(V/V) supernatants from PAM cells and 10%(V/V) supernatants from PMA-primed THP-1 cells,which were stimulated by 0,50,100,200 and 500 μg/ml SiO 2,could induce the proliferation of CHL or CCL-64 cells.The effects in 200 μg/ml and 500 μg/ml group were significantly higher than that in 0 μg/ml group,and there was a dose-effect relationship between the dose of SiO 2 and the degree of proliferation(CHL:r 1= 0.914, P< 0.05,r 2=0.980,P<0.01;CCL-64:r 1=0.975,P<0.01,r 2=0.956,P<0.01).10%(V/V) supernatants from PAM could suppress the proliferation of CHL or CCL-64 cells,the absorbency(A 570 nm) in 50 μg/ml SiO 2 groups(CHL:0.66±0.05, CCL-64:0.21± 0.04) were decreased significantly with the comparison to control(CHL: 0.74±0.04,CCL-64:0.26±0.04,P<0.01). Conclusion There is applicable perspective of using 5% (V/V) supernatnats form SiO 2-stimulated PAM cells and 10%(V/V) supernatants from SiO 2-stimulated PMA-primed THP-1 to induce CHL and CCL-64 cell proliferation to develop cellular model in vitro for studying silicotic fibrosis or neoplasm induced by silicon dioxide.

关 键 词：SIO2 肺纤维化 矽肺模型 矽肺 体外细胞模型 

分 类 号：R135.2[医药卫生—劳动卫生]