Effects Of Thy-1 DNA Methylation In Human Embryonic Lung Fibroblast Fibrosis Induced By Beryllium Sulfate

Objective By observing the Thy-1 DNA methylation changes in beryllium sulfate(Be SO4) stimulated the human embryonic lung fibroblast(MRC-5), Thy-1m RNA and protein changes, fibrosis related factors Collagen?,Collagen?, ?-SMA m RNA and protein changes to explore the effects of Thy-1 in beryllium sulfate(Be SO4) induced lung fibrosis.Methods Building the experimental model of cell culture in vitro, the final concentration were 1.0, 10.0, 100.0?mol/l of Be SO4(low?medium?high groups)?the control was untreated; the AZC antagonist group(10?mol/l AZC+ 10?mol/l Be SO4)and the TSA antagonist group(0.5?mol/l TSA+ 10?mol/l Be SO4). Collecting the cells after stimulated 24 h, 48 h, 72 h. RT-PCR determines the relative expression of Col?, Col?, ?-SMA and Thy-1 m RNA, The nested landed methylation specific PCR(nt-MSP) detects the Thy-1 DNA methylation level,Western-blot determined the relative expression of Col?, Col?, ?-SMA and Thy-1 protein.Results1. The relative expression of Col?, Col? and ?-SMA mRNA and protein in MRC-5 showed an increasing trend, by the increase of drug dose, it increased(P<0.05 or P<0.01); The low, medium, high groups of these three indicators in MRC-5 were all showing an increasing trend by the increase of expose time(P<0.05 or P<0.01); After the intervention of AZC and TSA,the relative expression of Col?, Col? and ?-SMA protein in Be SO4+AZC, Be SO4+TSA group were all lower than the medium group(P<0.05).2. In 24, 48, 72 h, the Thy-1 DNA methylation level in the medium, the high were all higher than the control. The Thy-1 DNA methylation level in low, medium, high groups were all showing an increasing trend by the increase of drug dose(P<0.05); After the intervention of AZC and TSA, in 48,72 h, the medium group was higher than the Be SO4+AZC and theBe SO4+TSA group(P<0.05).3. The relative expression Thy-1 m RNA and protein in three drug dose were all lower than those in control(P<0.05). After the intervention of AZC and TSA, the relative expression of Thy-1 protein level in the medium?high group were both higher than the mediuml group(P<0.05).Conclusion Fibrosis occurred in MRC-5 when stimulated with beryllium sulfate, in this process, Thy-1 DNA methylation level increased, Thy-1m RNA level reduced, may be one important mechanism lung fibrosis induced by beryllium sulfate, in the same time, it may also exist Histone acetylation.