ABSTRACT
Objective:To study the inhibition of astragaloside on the proliferation of human keloid fibroblasts. Methods: Com-pared with that of normal skin, the expression of transforming growth factor-β( TGF-β) and its transduction factors Smad in the human keloid fibroblasts was detected. The optimal concentration was screened by MTT after HFF-1 human skin fibroblast was infected with astragaloside at different concentrations. The mRNA expression of Smad2, Smad3, Smad4 and Smad7 in the fibroblasts was studied by using real-time. The protein expression of TGF-βRⅡ, Smad2, Smad3, Smad4 and Smad7 in the fibroblasts was detected by using Western blot. Results: Compared with that of normal skin tissue, the expression of Smad protein was significantly increased ( P 0. 05). The optimal concentration of astragaloside was 0. 5μg·ml-1 . The expression level of Smad2 protein in the two groups was significantly increased, and the level of Smad3 expression was significantly decreased (P<0. 05). Conclusion:Astragaloside can inhibit the formation of fi-broblast possibly through Smad2 over-expression and Smad3 inhibition in the TGF-β/Smad signal transduction pathway.
ABSTRACT
Objective To investigate the effect of small interfering RNA(siRNA)-expressing plasmid targeting connective tissue growth factor(CTGF) on the expressions of its mRNA and protein in cultured human keloid fibroblasts(hKFs).Methods Constructed siRNA-expressing plasmids targeting CTGF were transfected into hKFs by Dosper package.Then the transfected cells were treated with transforming growth factor 1(TGF-?1,10 ng/ml).Normal hKFs cells and those with TGF-?1 treatment served as normal control and control groups respectively.The expression level of CTGF mRNA and protein were detected by real-time fluorescent quantitative PCR(RQ-PCR) and Western blotting respectively.Results The expression level of CTGF mRNA was only 56.6% of that of the normal control(P