Effectiveness Of Artesunate Combined With Fractional CO₂ Laser Or 595nm Pulsed Dye Laser In Hypertrophic Scar And The Underlying Mechanism

Background and purpose:Hypertrophic scar(HS)is characterized by excessive proliferation,deposition and disorder of collagen,fibroblasts and extracellular matrix.The pathogenesis is not fully understood,which makes the clinical treatment difficult.Artesunate,carbon dioxide lattice laser and pulsed dye laser have been proved to be able to treat HS and other fibroproliferative skin diseases.However,there are no reports about artesunate combined with carbon dioxide lattice or pulsed dye laser in the treatment of HS.The aim of this study was to observe the effect of artesunate on human fibroblasts and combined therapy in the treatment of HS model,and to explore the effect of artesunate on TGF-β1,PCNA,BMP-7 and Fas,so as to better guide clinical work.Materials and methods:1.Methods:human HS fibroblasts were treated with artesunate.Cell proliferation and senescence were detected by WST-1 and β-gal methods.Cell apoptosis rate and growth cycle were detected by flow cytometry.2.Artesunate combined with carbon dioxide lattice laser or pulsed dye laser was used to treat HS rabbit ear model.He and VG staining were used to observe the histological changes and AA and Na values.3.CCK8(cell counting kit-8)was used to detect the viability of fibroblasts and a value at 450 nm in HS samples after treatment.4.ELISA was used to measure the content and ratio of type I and type III collagen in HS samples after treatment.5.The levels of TGF-β1,PCNA,BMP-7 and Fas were detected by immunohistochemistry,Western blot and RT-PCR.Result:1.Artesunate can reduce the proliferation ability of human HS fibroblasts,down regulate the proportion of S phase cells,reduce the proliferation rate and increase the apoptosis rate.2.Artesunate combined with carbon dioxide lattice laser or pulsed dye laser can significantly improve the appearance of HS rabbit ear model,reduce the activity of fibroblasts and the content of fibroblasts and collagen.3.Artesunate combined with carbon dioxide lattice laser can significantly reduce the contents of TGF-β1 and PCNA in HS samples and increase the contents of BMP-7 and Fas.4.Artesunate combined with pulsed dye laser can significantly reduce the contents of TGF-β1 and PCNA in HS samples and increase the contents of BMP-7 and Fas.ConclusionArtesunate can inhibit the proliferation of human HS fibroblasts.The combination of carbon dioxide lattice laser and pulsed dye laser can improve HS model,which may be related to the following factors:1.Artesunate can down regulate the proportion of S phase cells and up regulate the apoptosis rate of human fibroblasts;2.Artesunate combined with carbon dioxide lattice laser can reduce the content of fibroblasts and collagen in HS.3.Artesunate combined with pulsed dye laser can reduce the contents of TGF-β1 and PCNA,and increase the contents of BMP-7 and Fas.This study provides cytological and animal experimental data support for the possible mechanism of artesunate combined with carbon dioxide lattice laser or pulsed dye laser as a new therapeutic target for HS.