Objective: To screen the differential proteins on rats lung fibroblasts treated with or without transforming growth factor(TGF)-β1 by using a isobaric tags for relative and absolute quantitation(iTRAQ) profile and identify some differential proteins in rats exposed to silica. Methods: 5 ng·ml-1 TGF-β1 was used to induce fibroblasts for 24 h, and the differentially expressed proteins were screened out by iTRAQ combined with liquid chromatography-tandem mass spectrometry(LC-MS/MS) technology compared with the control group. The Scaffold software was used to verify the proteins and the bioinformatics analysis was performed with PANTHER online tool. Ten male Wistar rats with weight of (160±10) g were divided into control group and silicosis model group, 5 rats in each group. Silicotic rats were made by a HOPE MED esposure control apparatus[SiO2 concentration (50±10) g·m-3] for 24 weeks. VG staining was used to observe lung tissue morphology. The positive expression of collagen alpha-1(Ⅴ) chain(COL Ⅴ), vascular cell adhesion protein 1(VCAM), very low-density lipoprotein receptor(VLDLR), and syndecan-2(SDC2) were observed and measured by IHC staining and Western blotting. Results: A total of 1 648 proteins differentially expressed in response to TGF-β1 treatment and 196 proteins were found to be expressed at ≥ 1.2 fold relative to control. The silicotic lesions could be observed by VG staing in rats exposed to silica. The positive expression of COL Ⅴ, VLDLR, and SDC2 were observed in silicotic lesions by IHC stainging. Compared with control group, the expression of COL Ⅴ, VCAM, VLDLR and SDC2 increased in silicotic group(P<0.05). Conclusion: The results suggest that iTRAQ profile can be emplyed for identifying differential protein, COL Ⅴ, VCAM, VLDLR and SDC2 may have some roles on silicosis. |
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