Effect of Smurf1 on Fibrosis Formation in Hypertrophic Scar and its Molecular Mechanism
Objective:To explore the effect of Smurf1 on fibrosis process in hypertrophic scar formation and its molecular mechanism.Methods:Hypertrophic scar tissue and normal skin tissue samples of 12 patients who underwent hyperplastic scar resection in the department of burn and plastic surgery of the Second Affiliated Hospital of Air Force Medical University from June 2021 to June 2022 were collected.And the pathological examination was performed by HE and Masson staining.After sterile treatment of hypertrophic scar tissue,human hypertrophic scar fibroblasts(HSF)were isolated and cultured by tissue block method.HSF cells were divided into some groups according to the experimental scheme,(1)Smurf1 overexpression treatment group,control group 1(Con-1 group),empty vector group(Vector group)and Smurf1 overexpression group(OE-Smurf1 group),(2)Smurf1 interference expression treatment group,control group 2(Con-2 group),negative group(si-NC group)and Smurf1 interference expression group(si-Smurf1 group).Then pcDNA3.1 empty plasmid,pcDNA3.1+Smurf1 plasmid,si-NC and si-Smurf1 were transfected into HSF cells in Vector group,OE-Smurf1 group,si-NC group and si-Smurf1 group,respectively.Smurf1 mRNA expression level was detected by qRT-PCR.Protein expression level was detected by Western blot.Cell proliferation level was detected by CCK-8.Cell apoptosis level was detected by flow cytometry.Cell invasion level was detected by Transwell assay and cell migration level was detected by cell scratch assay.Results:The results of HE staining and Masson staining showed that compared with normal skin tissue,the dermis thickness of hypertrophic scar tissue increased significantly,and there were a large number of blue-dyed collagen fibers in the dermis,which were disordered and dense.Compared with normal skin tissue,the protein expression levels of TGF-β1,α-SMA,COL1,COL3,TβR-I,p-Smad3 and Smad7 in hypertrophic scar tissue were increased(P＜0.05),while the expression level of Smurfl was decreased(P＜0.05).Compared with Con-1 or Vector groups,the expression levels of TGF-β1,α-SMA,COL1,COL3,TβR-I,p-Smad3 and Smad7 proteins in HSF cells of OE-Smurfl group were decreased(P＜0.05),and the proliferation,invasion and migration levels of HSF cells were decreased(P＜0.05),the apoptosis level was increased(P＜0.05).Compared with Con-2 group or si-NC group,the expression levels of TGF-β1,α-SMA,COL1,COL3,TβR-I,p-Smad3 and Smad7 proteins in HSF cells of si-Smurfl group were increased(P＜0.05),and the proliferation,invasion and migration levels of HSF cells were increased(P＜0.05),the apoptosis level was decreased(P＜0.05).Conclusion:Smurfl may inhibit the fibrotic process of hypertrophic scars by inhibiting TGF-β1/Smad pathway.
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