ITGB3对增生性瘢痕成纤维细胞生物学行为及TGF-β1/Smad2信号通路的影响OA

Objective To explore the role and mechanism of Integrin β3(ITGB3)in Hypertrophic scar(HS).Methods A total of 30 hyperplastic scar tissue specimens from patients diagnosed with HS in the Department of orthopedic in Xi'an Central Hospital from May 2022 to May 2023 were collected,and 30 normal skin tissue specimens from surgically removed patients were collected during the same period.The degree of lesion and fibrosis in hypertrophic scar tissue were detected by HE staining and Masson staining.ITGB3 expression in hypertrophic scar tissue was detected by immunocytochemistry.Human hypertrophic scar fibroblasts(hHSFs)was divided into control group,sh-NC group(hHSF cells were infected with sh-NC),and sh-ITGB3 group(hHSF cells were infected with sh-ITGB3),and SRI-011381 group(hHSF cells were infected with sh-ITGB3 and treated with TGF-β1 activator SRI-011381).The cell proliferation was detected by CCK-8 method and EdU fluorescence labeling method.The cell apoptosis was detected by flow cytometry,and cell migration was detected by cell scratch assay.ITGB3 gene expression levels in tissues and cells were analyzed by qRT-PCR.while protein expression levels of ITGB3,Collagen I,α-SMA,MMP-2,TIMP-2,TGF-β1,p-Smad2 and Smad2 in tissues and cells were analyzed by Western blot.Results The boundary between epidermis and dermis was obvious in normal skin tissue,and the collagen fibers were arranged neatly with uniform density.The dermis in HS tissue was obviously thickened and there were a large number of disordered and irregularly arranged collagen fibers.Compared with normal skin tissues,mRNA and protein expression levels of ITGB3 in HS tissues were increased(P＜0.05),protein expression levels of TGF-β1,Collagen I,α-SMA and TIMP-2 and p-Smad2/Smad2 ratio were increased(P＜0.05).And the expression level of MMP-2 protein was decreased(P＜0.05).Compared with hSF cells,the mRNA and protein expression levels of ITGB3 in hHSF cells were increased(P＜0.05).Compared with the control group or sh-NC group,the mRNA and protein expression levels of ITGB3 in sh-ITGB3 group and SRI-011381 group were decreased(P＜0.05).Compared with control group or sh-NC group,the cell proliferation level and relative cell migration rates in the sh-ITGB3 group were decreased(P＜0.05),while the cell apoptosis level was increased(P＜0.05),the TGF-β1,Collagen I,α-SMA,TIMP-2 protein expression level and p-Smad2/Smad2 ratio were decreased(P＜0.05),MMP-2 protein expression level was increased(P＜0.05).Compared with sh-ITGB3 group,the cell proliferation level and relative cell migration rates in the SRI-011381 group were increased(P＜0.05),while the cell apoptosis level was decreased(P＜0.05),the TGF-β1,Collagen I,α-SMA,TIMP-2 protein expression level and p-Smad2/Smad2 ratio were increased(P＜0.05),MMP-2 protein expression level was decreased(P＜0.05).Conclusion ITGB3 is highly expressed in HS tissues,which may promote the hHSF cells proliferation and migration and ECM deposition through activation of the TGF-β1/Smad2 signaling pathway,and inhibit the apoptosis of hHSF cells,thus promoting the formation of hypertrophic scars.