Experiment Study On Suppression Of TGFβ1 In Hypertrophic Scar By Vector-Based Small Haprin RNA

Pathological scar(Hypertrphic scar and Keloid)will from when extracellular matrix(ECM)abnormally deposits after skin destruction.It can demolish our body surface perfection,even bring us function disturbance,and it is always important clinic topic.The cicatrix fibroblast is the key effector cell in the scarring.The dysfunctional fibroblasts is known as the reason of abnormal scar formation recently,and transforming growth factor(TGFB)is the most intimate factor in the over-scarring.TGFβ1 can increase ECM synthesis including collagen/fibronectin and hyaluronic acid in vitro experiment. Synthetical TGFβ1 can adjust precollagenⅠ/Ⅲgene express in the scar tissue. Because the relation ship between TGFβ1 and scarring exist causal relation,the treatment against TGFβ1 maybe the effective method to cure cicatrix.At present,development of study on gene therapy of abnormal scar has been improved.Many scholars try inhibiting TGFβ1 by antisense oligonucleotides and ribozyme,which have low transduction efficiency and transit time.RNA interference(RNAi)technique emergence provides a new way to inhibit gene expression.RNAi is the most transcription gene silence induced by dsRNA,which can silence gene specifically and result in gene functional incapacitation or reduction.RNAi has specificity,persistence and sensitivity.However there are less numerous studies about cure hypertrophic scar by RNAi in the world.Objective:To explore the use of specific small hairpin RNA(shRNA)to suppress TGFβ1 gene expressions in human hypertrophic scar fibroblast,and investigate the effect of TGFβ1 shRNA on typesⅠandⅢprocollagen expression.Methods:TGFβ1 specific shRNA expression vector were constructed and introduced to the eight primary hypertrophic scar fibroblasts.Expression of TGFβ1 mRNA was assessed by Real-Time quantitative reverse transcription polymerase chain reaction.The TGFβ1 protein level in the culture supematant was determined by sandwich enzyme-linked immunosorbent assay.Results:The expression of TGFβ1 was upregulated significantly in hypertrophic scar fibroblasts.TGFβ1 expression after twenty four hours and forty eight hours in TGFβ1 pSUPER plasmid vector-mediated TGFβ1 shRNA groups were obviously downregulated when compared to nonspecific pSUPER and transfection reagent alone group(P＜0.05).No significant difference was found between nonspecific pSUPER vector plasmid-mediated TGFβ1 group and transfection reagent alone group(P＞0.05).The expression of TGFβ1 signaling downstream genes in TGFβ1 pSUPER plasmid vector-mediated TGFβ1 shRNA group were also obviously downregulated when compared to t nonspecific pSUPER and transfection reagent alone groups(P＜0.05).Conclusions:TGFβ1 specific shRNA can significantly inhibit the expression of TGFβ1 in hypertrophic scar fibroblasts.These results suggest the possible application of TGFβ1 specific shRNA in preventing hypertrophic scar in patients.