Study Of Inhibition Of Dihydroartemisinin To The Humanurethral Fibroblasts Cell Proliferation

Objectives:The incidence of urethral stricture post urethroplasty or urethral injury is quite high, butthere is still no effective way to clinically prevent it at present. The patients who are always ingreat pain have been trapped in the vicious circle of urethral stricture anddilatation/surgery. The urethral scar tissue is the byproduct of wound repair, it usually meanswound healing. However, once the scar tissue overgrows, urethral stricture happens.Nowadays, it has been discovered that urethral stricture tissue contains typical hyperplasticscar. But it is indefinite for the influencing factor of hyperplastic scar in urethral, so there isno effective measure to prevent urethral stricture. In the research of scar formation of skinscar, we find overgrowth and collagen I secretion of fibroblast during agglutination of rawsurface, which we think is the reason of scar. Fibroblast is the overriding repair cell.Artemisinin is a drug with independent intellectual property in China, which we strong onresearch and development. Artemisinin was found have the effect on inhibition to thefibroblasts cell proliferation in scar recently. Through the study of inhibition ofdihydro-artemisinin to the human urethral fibroblasts cell proliferation, we want find themechanism of inhibition of dihydro-artemisinin to the human urethral fibroblasts cellproliferation, find a new of prevention and treatment of urethral stricture, provid theorybase for prevention and treatment of urethral stricture in clinic.Methods:1.Tissue adherent method was used in primary culture of keloid fibroblasts, RPMI-1640medium containing10%fetal calf serum,37℃，5%CO2，saturated humidity training, describethe growth curve, mitotic index, morphology and vimentin expression.2.Using MTT assay the IC50of dihydroartemisinin to keloid fibroblasts, Using TUNELassay apoptosis of keloid fibroblasts after dihydroartemisinin effect, While using colorimetricdetermination of caspase-3,8and9after the role in the activation of dihydroartemisininsituation, to determine whether dihydroartemisinin lead to the caspase activation of keloidfibroblasts, And understand which of the paths through the promotion of apoptosis.Results:1.The success rate of keloid fibroblast cell culture was75%(6/8); the first generation need50to66days, and5to10days every generation afterward, grow well after the passage;Long spindle-shaped cells, vimentin expression was positive.2.The IC50of dihydroartemisinin to keloid fibroblast was145μmol/L. By TUNEL assayto150μmol/L dihydroartemisinin role in keloid fibroblasts can see a clear apoptotic cells.Detected by colorimetric caspase-9and caspase-3activation, but not caspase-8activation.Conclusions:1. Keloid fibroblasts cultured by tissue adherent method can be used in vitro, and toprovide material basis for further experiments.2. A certain concentration of dihydroartemisinin can promote apoptosis in keloidfibroblasts, and start the apoptosis process through the activation of caspase-9, and thenactivation of caspase-3.