Expression And Function Of WD Repeat Proteins WDR26in Dermal Fibroblasts Of Systemic Sclerosis

Objective:We investigated the expression of WDR26at protein level in dermal fibroblasts of systemic sclerosis patients and healthy controls, and the expression of WDR26in the dermal fibroblasts with TGF-β stimulation.Methods:①Western-blot analysis was used to test the expression of WDR26in dermal fibroblasts of systemic sclerosis patients and healthy controls, and the expression of WDR26in healthy dermal fibroblasts with TGF-β stimulation.②Real-time PCR was used to detect the expression of WDR26mRNA in SSc and healthy dermal fibroblasts after the TGF-β stimulation at different concentrations(0,0.1,1.0,5.0,10.0ng/ml) and different time durations (0,6,12,24,48h).③Real-time PCR was used to investigate the effect of WDR26in TGF-β induced production of extracellular matrix (such as COL1A1,FN) at transcript levels in the fibroblasts with forced overexpression and inhibition of WDR26. The expression of SMAD3mRNA was also tested in the fibroblasts with forced overexpression and inhibition of WDR26.Results:①Our data revealed that the WDR26protein expression level was higher in SSc dermal fibroblasts than in healthy control. And TGF-β up-regulated WDR26protein expression in healthy dermal fibroblasts.②After the stimulation with TGF-β, WDR26mRNA expression was elevated at first and then reduced with the rising TGF-β concentrations and the lasting simulation time.③The data revealed that the elevated expression of extracellular matrix COL1A1mRNA simulated by TGF-β had been inhibited in the WDR26overexpression fibroblasts; the elevated expression of extracellular matrix COL1A1mRNA simulated by TGF-β has been promoted in the WDR26suppressed fibroblasts. SMAD3mRNA expression was inhibited in both WDR26suppressed and overexpressed fibroblasts. SMAD3mRNA expression was elevated at first and then reduced with the elevating expression of WDR26in fibroblasts.Conclusion:①WDR26protein expression level is higher in SSc dermal fibroblasts than in healthy control.②TGF-β can induce the WDR26expression in dermal fibroblasts.③WDR26inhibits the expression of COL1A1mRNA in dermal fibroblasts stimulated by TGF-β and releases the fibrosis induced by TGF-β...