The inhibitive effect of PTEN protein on secretion of collagen Ⅳ and fibronectin by stimulation of by TGF-?_1 in rat / 解放军医学杂志

ABSTRACT

Objective To investigate the role of PTEN protein in secretion of collagen Ⅳ and fibronectin after stimulation of transforming growth factor beta 1(TGF-?1) from renal fibroblasts of rat in vitro.Methods The cultured rat renal fibroblasts were transfected with the reconstructed adenovirus containing PTEN or adenovirus only containing green fluorescence protein(GFP).The fibroblsts were treated in four mannerscontrol group with no added treatment,TGF-?1 group with TGF-?1 stimulation,PTEN+ TGF-?1 group with TGF-?1 stimulation after Ad-PTEN transfection,and GFP+ TGF-?1 group with TGF-?1 stimulation after Ad-GFP transfection.Invert fluorescent microscope was used to detect the GFP expression,meanwhile the PTEN mRNA was determined by RT-PCR method.36h after the transfection,TGF-?1 was added into the culture medium in a concentration of 10ng/ml.After another 24h,ELISA method was used to evaluate the level of collagen Ⅳ and fibronectin.Results The expressions of both GFP and PTEN mRNA increased obviously after the rats' renal fibroblasts were transfected with adenovirus.The secretion of collagen Ⅳ and fibronectin increased significantly in both TGF-?1 group and GFP+ TGF-?1 group compared with that in control group,and decreased markedly in PTEN+ TGF-?1 group compared with that in both TGF-?1 group and GFP+ TGF-?1 group(P