二氧化硅对肺成纤维细胞连接蛋白Cx43表达的影响  

Study on the effect of silicon dioxide on connexin 43 expression in pulmonary fibroblast

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,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292-294,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　通过研究SiO2 对连接蛋白 (Cx)Cx4 3表达的调节 ,了解SiO2 抑制肺成纤维细胞细胞间隙连接通讯 (GJIC)功能的机制。方法　以不同剂量SiO2 刺激SD大鼠肺泡巨噬细胞 (PAM)及佛波酯(PMA)诱导分化的THP 1(具有PAM特性的人血单核细胞株 )细胞培养上清液 ,作用于中国仓鼠肺成纤维细胞 (CHL) ,采用双抗体夹心酶联免疫吸附测定法 (ELISA)相对定量CHL细胞Cx4 3的含量 (以A492nm表示 ) ,用四唑盐 (MTT)法测定其增殖效应 (以A570nm表示 )。结果　在 0、5 0、10 0、2 0 0、5 0 0 μg/mlSiO2 浓度范围内 ,SiO2 剂量与SiO2 刺激的PAM和PMA primedTHP 1上清液诱导的增殖效应呈正相关 (r1=0 .914 ,P <0 .0 5 ;r2 =0 .980 ,P <0 .0 1)。SiO2 刺激的PAM上清液促进CHL细胞Cx4 3蛋白的表达 ,5 0、10 0、2 0 0、5 0 0 μg/mlSiO2 剂量组Cx4 3的A492nm值分别为 ( 0 .171± 0 .0 2 7、0 .161± 0 .0 4 1、0 .2 5 7± 0 .0 4 0和 0 .2 0 8±0 .0 2 6) ,与SiO2 0 μg/ml对照组 ( 0 .10 5± 0 .0 0 8)比较 ,差异均有显著性 (P <0 .0 5 )。SiO2 刺激的PMA primedTHP 1细胞上清液对CHL细胞Cx4 3蛋白表达水平无明显影响。结论　SiO2 刺激PAM分泌活性因子进而抑制肺成纤维细胞GJIC功能的调节机制可能发生在Cx的翻译后水平。Objective To explore the possible mechanism on down-regulation of gap juactional intercellular communication(GJIC) of fibroblast stimulated by silicon dioxide. Methods The pulmonary alveolar macrophage(PAM) or the phorbol 12-myristate 13-acetate(PMA)-primed THP-1 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 cells were incubated with the prepared supernatants for 24 hours.After incubation,the relative quantitation of Cx43 in CHL cells were detected using sandwich enzyme-linked immunoassay(ELISA),and the proliferation of CHL cells was detected using MTT assay(to show as the absorbency,A 570 nm). Results In the ranges of 0,50,100,200 and 500 μg/ml silicon dioxide concentrations,the proliferation of CHL cells induced by the supernatants from PAM and PMA-primed THP-1 and silicon dioxide concentrations were fitted well in the dose-response relationship (r 1= 0.914,P<0.05;r 2=0.980,P<0.01).The supernatants from PAM could promote the expression of Cx43,and the absorbency(A 492 nm) in 50,100,200,500 μg/ml silicon dioxide groups(0.171± 0.027,0.161±0.041,0.257± 0.040,0.208±0.026,respectively) were significantly higher than that in control(0.105±0.008,P<0.05).The supernatants from PMA-primed THP-1 stimulated by silicon dioxide had no effect on the expression of Cx43.T5'HZ] Conclusions SiO 2 may control the post-translational process of Cx43 to inhibit GJIC function of fibroblast. Conclusions SiO 2 may control the post-translational process of Cx43 to inhibit GJIC function of fibroblast.

关 键 词：SIO2 连接蛋白CX43 ELISA 肺纤维化 成纤维细胞 二氧化硅 

分 类 号：R135.2[医药卫生—劳动卫生]