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
AIM:To investigate whether Smad pathway participates the process of extracellular signal regulated kinase (ERK) induced the proliferation of vascular smooth muscle cells (VSMCs). METHODS: Human umbilical artery smooth muscle cells (hUASMCs) were divided into four groups: control group, PDGF (platelet derived growth factor) group, ERK blocking agent group and PDGF+ERK blocking agent group. MTT assay was used to detect the proliferation of hUASMCs (A value). Immunohistochemical technique was used to detect the expression of PCNA, phosphorylated ERK (p-ERK) and phosphorylated Smad2/3 (p-Smad2/3) protein in hUASMCs. The expression of Smad2/3 mRNA in hUASMCs was detected by RT-PCR. RESULTS: The proliferation of hUASMCs and the expression of PCNA, p-ERK and p-Smad2/3 proteins in hUASMCs in PDGF group were increased obviously than those in other groups (P