Effects of Ursolic Acid on the Biological Function of Scar Fibroblasts and the TGF-β1/Smads Pathway
The aim of this study is to investigate the effects of ursolic acid on the biological function of scar fibroblasts and the TGF-β1(transforming growth factor-β1)/Smads pathway.By culture with different doses of ur-solic acid(0,10,20,30,40,50,60 μmol/L),cell survival rate of HSFB(hypertrophic scar fibroblast)in logarithmic growth phase was firstly detected by MTT methods.After that,five experimental groups,named as control group(without ursolic acid adding in culture medium),L-ursolic acid group(with 20 μmol/L ursolic acid adding in cul-ture medium),M-ursolic acid group(with 40 μmol/L ursolic acid adding in culture medium),H-ursolic acid group(with 60 μmol/L ursolic acid adding in culture medium),H-ursolic acid+SRI-011381 group(with 60 μmol/L ursolic acid and 10 μmol/L TGF-β1/Smads pathway activator SRI-011381),were selected for the further study.Apoptosis of HSFB when culture in the above groups was determined by flow cytometry and TUNEL staining.Transwell ex-periment was applied to measure migration and invasion abilities of HSFB.Western blot was applied to determine the expression of TGF-β1/Smads pathway proteins(TGF-β1,p-Smad2,p-Smad3,Smad4),fibrotic proteins(Col 1A1,Col 3A1,α-SMA),and apoptotic proteins(Bax,Bcl-2)in each group.The result showed that with the con-centration of ursolic acid increased in culture medium,the survival rate of HSFB gradually decreased in a dose-dependent manner(P＜0.05).As compared with the blank control,the apoptosis rate,proportion of TUNEL positive cells and expression of Bax protein in HSFB in the L-ursolic acid group,M-ursolic acid group,and H-ursolic acid group were all increased(P＜0.05),while the numbers of migrating cells,invading cells,and the expression of fi-brotic marker proteins(Col 1A1,Col 3A1,α-SMA,Bcl-2,TGF-β1/Smads pathway related proteins)of HSFB were all reduced on the contrary(P＜0.05).Those degree of change in H-ursolic acid group was more obvious than that of L-ursolic acid group(P＜0.05).Furthermore,the apoptosis rate,proportion of TUNEL positive cells and expres-sion of Bax protein of HSFB in H-ursolic acid+SRI-011381 group were reduced(P＜0.05),while the numbers of migrating cells,invading cells,and the protein expression of fibrotic marker proteins(Col 1A1,Col 3A1,α-SMA),Bcl-2,TGF-β1/Smads pathway related proteins in H-ursolic acid+SRI-011381 group were increased(P＜0.05),as compared with that of the H-ursolic acid group.As a reminder,ursolic acid can effectively inhibit the prolifera-tion,invasion,migration,and fibrosis(Col 1A1,Col 3A1,α-SMA)of HSFB,and further induce HSFB's apoptosis.All the biological processes of HSFB that mediated by ursolic acid may be related to inhibiting the activation of TGF-β1/Smads signaling pathway.
hypertrophic scarsfibroblastsTGF-β1(transforming growth factor-β1)/Smadsbiological function
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维普
