Fucosylation Influences Extracellular Matrix Accumulation in Tgf-β-stimulated Hk-2 Cells
|School||Dalian Medical University|
|Keywords||α1，6-Fucosyltransferase Fucosylation extracellular matrix Matrix metalloproteinase renal interstitial fibrosis|
Objective. To investigate the effect ofα-1,6 fucosyltransferase (Fut8) on the extracelluar matrix accumulation induced by transforming growth factor-β1 (TGF-β1) in human renal proximal tubular cells(HK-2).Methods. The cultured HK-2 cells were divided into six groups randomly, (1)the normal control group(CON group) as a negative control group, HK-2 cells were cultured in the normal media. (2)Mock group, HK-2 cells were transfected with scrambled siRNA. (3)TGF group, cells were treated with 10ng/ml TGF-β1. (4)TGFM group, cells were transfected with scrambled siRNA and then treated with 10ng/ml TGF-β1. (5)Fut8-siRNA group, cells were transfected with Fut8-siRNA. (6)TGFF group, cells were transfected with Fut8-siRNA and then treated with 10ng/ml TGF-β1. The expression changes of Fut8 were examined by western blot, the apoptosis were examined by flow cytometry, protein levels of MMP-2,3,9 and TIMP-1 were examined by western blot, ECM proteins, including collagen typeⅠ,Ⅰ,Ⅰwere examined by immunofluorescence, fibronectin and laminin were examined by Real time-PCR.Results. The expression of Fut8 were increased in HK-2 cells stimulated by TGF-β1(P<0.05), were decreased(P<0.05) in HK-2 cells transfected with Fut8-siRNA. Fut8-siRNA increased the expression of MMP-2,3,9, and inhibited apoptosis, ECM accumulation and the expression of TIMP-1 in HK-2 cells stimulated by TGF-β1.Conclusion: Fut8-siRNA block the level of fucosylation, up-regulate matrix metalloproteinases and down-regulate extracellular matrix protein accumulation in HK-2 cells stimulated by TGF-β1, then suppress interstitial fibrosis.