Expression And Significance Of IGF, VEGF, Decorin And Versican In The Pathological Scars

[Objective]This study was designed to explore the expression and significance of growth factors(IGF,VEGF) and extracellular matrix(decorin,versican) during formation of scars,especially keloids.Reveal the mechanism to provide basic theory for clinical precaution and treatment to scars at molecular and pathological level.[Method]1.The f ibroblasts of keloid,normal scar and normal skin were cultured in vitro.Their difference of morphology,ultrastructure and cell activity will be observed by light microscopy,transmission electron microscopy,flow cytometry;2.Total RNA Will be extracted from paraffin embedded tissue(keloids and normal skin) and expression of VEGF,decorin and versican will be detected by PCR;3.By immunohistochemistry,immunofluorescence and confocal laser microscopy,ELISA detection,real-time fluorescence quantitative PCR and the westen blot,the expression of IGF,VEGF,decorin and versican will be detected and analyzed in the fibroblasts;4.Hypertrophic scar models were made on pig back in vivo by different concentration Salvia yunnanensis C.H.Wright injection, detected by HE staining,Masson's staining,transmission electron microscope and in situ hybridization.Using these experimental analyse,to study the biological behavior and pathological changes of hypertrophic scar fibroblasts affected by Salvia yunnanensis C.H.Wright. [Result]1.The result of Light microscopy showed:there are no significant differences in morphology of fibroblasts of Keloids,scars and nomal skin.The result of transmission electron microscopy showed there are more rich with rough endoplasmic reticulum and mitochondria in fibroblasts of keloids than that of scars and nomal skin.The result of flow cytometry indicated that fibroblasts of keloid was higher in S phase percentage,that means the better the activity of cells;2.The result of PCR with total RNA from paraffin-embedded tissue show:compared with normal skin,keloid were significantly higher in VEGF expression,decorin expression was significantly reduced;and the four subtypes of versican expression in keloids compared with normal skin,there is no significant difference;3.fibroblasts immunofluorescence showed that:in keloid fibroblasts,the expression of IGF,VEGF and versican were higher,while in normal skin in a higher decorin expression;4.The result of ELISA with fibroblasts culture supernatant show:compared with normal skin,keloid higher concentrations of IGF and VEGF,compared with keloids,normal skin decorin concentrations higher;5.The result of Real-time fluorescence quantitative real-time quantitative PCR with RNA from fibroblast show:compared with normal skin,keloid in the expression of VEGF and 2-fold increase,decorin expression decreased about 5 times;and versican of the four subtypes were expressed in keloid and have higher than normal skin,expression from high to low is the V1, V3,V0,V2,respectively,10 times,8 times,4 times,3 times higher;6.The result of westen blot show:in the protein level,compared with normal skin, VEGF in keloid is significantly elevated;compared with keloids,expression of decorin was higher in normal skin;7.In vito(pig back scar model):Compared with the control group,by pathological staining,the experiment group were less fibroblasts and the collagen density reduced and arranged in regulation.The spaces between fiber bundles were increased.By TEM,compared with the control group,there were less fibroblast than fibro and undeveloped rough endoplasmic reticulum.Heterochranatins were very abundance and an arrangement of collagen fibers which were orderly.In part rough endoplasmic reticulum were over expand and have the characteristic of posterior fibroblast.The expression of decorin mRNA was detected by in situ hybridization:The expression of decorin mRNA was rich in the normal skin dermis.The lower expression of decorin mRNA was found after one more months with Salvia yunnanensis C.H.Wright.injection,but the control group were not found.[Conclusion]In the skin wound healing,overexpression of IGF,VEGF,and versican may promote scars formation,while low expression of decorin leads to scar formation;Salvia yunnanensis C.H.Wright can decrease fibroblasts growth, proliferation and collagen synthesis.