The Preliminary Experimental Study, The Nuclear Transcription Factor Egr-1, Nf-¦Êb In Targets For The Treatment Of Silicon Pulmonary Fibrosis

ObjectiveTo study the molecular mechanisms of up-regulating TNF-α and TGF- β1 in macrophages by silica stimulation and the effects of the signal pathway mediated by Egr-1 and NF-kappa B on the occurrence of silicosis. Methods(1) RAW264.7 cells were divided into blank control group, silica stimulation group, egr-1 antibody + Si 62 group, NF- K B antibody + SiO2 group and IgG+ SiO2 group, and pretreated with Egr-1 and NF- K B antibody respectively to block their intrinsic effects , then stimulated with silica and added egr-1 and NF- K. B antibodies respectively for one hour ;(2) RAW264.7 cells were divided into blank control group, silica stimulation group, sense egr-1 +SiO2 group ,antisense egr-1 + SiO2 group, sense NF- K B + SiO2 group, antisense NF- K B + SiO2 group and lipofectin + SiO2 group, and transiently transfected with Egr-1 and NF- K B oligonucleotides respectively, then were treated with silica for one hour;(3) TNF- α protein level in cell supernatants was measured using enzyme-linked immuoadsorbent assay (ELISA), and TGF- β1 protein expression was detected by immunocytochemistry. At the same timetheir mRNA expressions were monitored by RT-PCR. Results(1) TNF- α and TGF-β1 gene expressions were significantly higher in the silica stimulation group than that in the blank group. Compared to RAW264.7 cells untreated with specific antibody under silica stimulation, RAW264.7 cells treated with 10ug/ml of egr-1 or NF- K B antibody could reduce the expressions of proteins and mRNAs of TNF-α and TGF- β1 (P<0.05).(2) Compared to RAW264.7 cells under silica stimulation in the sense groups and untransfected groups, the expressions of proteins and mRNAs of TNF- α and TGF- β1 obviously reduced (P<0.05) in the antisense groups.ConclusionThe activation of Egr-1 and NF- K B might involve in the expressions of proteins and mRNAs of TNF- α and TGF- β1 induced by silica in RAW264.7 cells. The antibodies and antisense oligonucleotides of Egr-1 and NF- K B might inhibit the expressions of TNF- αa and TGF- β1 The findings suggest that these antibodies and antisense oligonucleotides might become a new therapeutic choice for pulmonary fibrosis in the future.