Effect Of Transforming Growth Factorβ In The Happening Of Recurrence Urethral Stricture

Background and Objectives:Patients suffering from urethral stricture have greatly increased with the intensive happening of all kinds of trauma, venereal disease and extensive development of endoscope for the past few years. As a result, more and more patients have to meet the recurrence urethral stricture. Clinician can do nothing but to do with anatomic structure for those recurrence urethral stricture sufferers.We all know that there is still no effective methods to stop it happening and the results for its curing are always making us unsatisfied, which are caused that we have know little about the happening of recurrence urethral stricture. Now it has been discovered that the tissure of recurrence urethral stricture is made of typical hyperplastic scar.Repair in trauma depends on the interaction between the cell and extracellular matrix. Fibroblast is the most important reparing cell, which immigrates to raw surface with the contribution of some cell chemotatic factor and sends out plenty of extracellular matrix. Hyperplasia of fibroblasts and increasing of ECM synthesis all lead to ECM accumulating in raw surface and result in the formation of hyperplastic scar. Now it has been proved that the happening of hyperplastic scar has greatly correlation with transformimg growth factor beta (TGF-β), which is powerful chemotatic factor to fibroblastic. TGF-βis a supergene family, which has three types in mammal, called TGF-β1～TGF-β3 and they have different disposition and expression in tissue. They have different contribution in the happening of hyperplastic scar. So, we come to study the characteristic of recurrence urethral stricture histopathology and hope to find out relationship with TGF-β.Methods:To collect recurrence urethral stricture tissues and normal urethra tissues as studying samples. HE dyeings are made to observe the urethral tissue structure. Collagen fiber and elastic fibers dyeings are made to compare the discrepancy between recurrence urethral stricture tissues and normal urethra tissues. Immunohistochemistry dyeings are used to study the disposition and expression of the three different type TGF-βin recurrence urethral stricture tissues. MetaMorph image analysises are used to carry out semiquantitative analysis for every group slices.To cultivate recurrence urethral scar fibroblast, and to observe the effect of TGF-β1 on its biological activity. To design and synthesize plus sense oligodeo xynucleotide (PS-ODN), antisense oligonucleotides(ASODN), random oligonucleotides(RDODN). Control group, Lipofectamine group, PSODN group, RDODN group and ASODN group are designed. Except control group, the rest are all performed transient transfection fibroblast mediated by lipofectamine. And to observe the effect of ODN on the proliferation of fibroblasts, TGF-β1 mRNA expression and the secretion of collogen1.Main results and conclusions:1. To compare with normal urethra tissue, recurrence urethral stricture tissues' feature:lumina nan'owing, irregular, urethra epithelium mucosae attenuation, even vanishing, below membrana mucosa plenty of fibroblasts can be seen.2. Dyeing of collagen fibers and elastic fibers demonstrate normal cavernous body of urethra tissues are replaced by plenty of fiber tissues, a lot of collagen fibers existed and structure confused, elastic fibers obviously decreased. Contusion: hyperplasia sear in the recurrent urethral stricture, disproportion of collagen fibers and elastic fibers which make tissue stiff and short of elasticity, as a result, barrier for urinating is coming.3. The intensity of the staining for TGFβ1 in urethra scar specimen was stronger. TGFβ2 and TGFβ3 was weaker than that for TGFβ1. Excessive expression of TGFβ1 which existed in numerous fibroblasts and endothelial cells could possibly account for the formation of hyperplasia scar.4. After transient transfection fibroblast mediated by lipofectamine, RT-PCR semiquantitatives find that ASODN can deeply depress TGF-β1mRAN expression of fibroblasts.5. After transient transfection fibroblast mediated by lipofectamine, the results of Elisa detection exhibit that ASODN can obviously inhibit the synthesis and secretion of collogenl from fibroblasts.6. After transient transfection fibroblast mediated by lipofectamine, MTT experiments discovery that ASODN can restraint the proliferation of fibroblasts.