TGF-β1 Affecting The Expression Of Smads Protein And Collagen In Human Atrial Fibroblasts And Study Of Intervention Of Atorvastatin

【Objective】By using TGF-β1 and atorvastatin to interfere cultured atrial fibroblasts,to observe the changes of expression of typeⅠcollagen,Smad2,Smad4 and Smad7 and to Study the mechanism of fibrosis in patients with atrial fibrillation,and of antifibrotic of atorvastatin.【Methods】Through cardiac surgery patients with Sinus rhythm,right atrial appendage organization 30mg was achieved ,and cardiac fibroblasts were isolated and cultured,observing the cardiac fibroblast morphology and growth state in the inverted microscope;Cells were identified by immunocytochemistry.After passage in cell culture,cardiac fibroblasts were randomly divided into control group and experimental group; After the intervention of TGF-β1 and atorvastatin, the proliferation of cardiac fibroblast was observed in the MTT method;Changes of Smad2,Smad4,Smad7 and collage typeⅠwere observed by RT-PCR and Western blot techniques.【Results】(1) Human atrial cardiac fibroblasts were successfully cultured in the tissue culture method; Morphology of the cardiac fibroblasts were observed in the inverted microscope.Myocardial fibroblasts growed Slowly, abaut three or five days climbing from the edge of the tissue , ten or twelve days becoming fusion state, and arranging closely,and some overlaping; It was spindle-shaped,larger,transparent in the cytoplasm,and it was much larger nucleus that was oval-shaped, usually with 1 or 3 nucleus and non-spontaneous beat.Identification of immunocytochemical staining showed that cardiac fibroblasts showed strongly positive reaction to vimentin and negative to factor VIII,which showed the purity of cells cultured was more than percent 95. (2) MTT showed that TGF-β1 promoted cardiac fibroblasts growth, in the 10ng/ml achieving the best in the proliferation,and atorvastatin inhibited cardiac fibroblast growth, at 10μmol/L concentration reaching the maximum inhibition.(3) Compared with the control group, In 1ng/ml, 10ng/ml TGF-β1 intervention,The mRNA and protein expression levels of Smad2, Smad4,and collagen typeⅠwere significantly higher (P <0.05) in cardiac fibroblasts,but Smad7 significantly lower (P<0.05). with 10ng/ml TGF-β1 group compared, in 10ng/ml TGF-β1,and 10μmol/L atorvastatin joint intervention group and 10μmol / L atorvastatin group, the mRNA and protein expression levels of Smad2, Smad4, collagen typeⅠon cardiac fibroblasts were significantly lower (P<0.05), however,Smad7 was significantly higher (P<0.05).【Conclusion】Using tissue culture method human atrial cardiac fibroblasts were successfully cultured; TGF-β1 promoted the growth of cardiac fibroblasts,and atorvastatin inhibited the proliferation of cardiac fibroblasts; TGF-β1 increased expression of collagen typeⅠin atrial cardiac fibroblasts through increasing Smad2, Smad4 expression,yet decreasing Smad7 expression; Atorvastatin reduced the expression of collagen typeⅠon cardiac fibroblasts, probably by way of TGF-β1/Smad.