Effects Of Prostatic Epithelial Cells And CKIP-1 Leves On The Expression Of TGF-β₁ And α-SMA In Fibroblasts Of Prostatic Urethral Wounds

Objective:The study showed the amount of scar tissue in canine’s prostatic urethra wound were less than in bladder neck wound after wound repair.In this stydy we observed the histopathologic characteristics and evaluated the expression of transforming growth factor beta1（TGF-β₁）and casein kinase 2interaction protein 1（CKIP-1）in the process of wound repair after two-micron laser resection of the prostate and the bladder neck through establishment of canine animal model.To reveal the effects of CKIP-1 level of prostatic epithelial cells on the expression of TGF-β₁ andα-SMA in fibroblasts of prostatic urethral wounds in vitro,then we constructed CKIP-1 overexpressed human prostate epithelial cells（BPH-1cells）,and co-cultured BPH-1 cells and human fibroblast（HFF-1 cell）.By this way,we explored the probable causes of formation of scar tissue（less scar healing）after prosthetic urethral wound repair.Methods:（1）Animal model of two-micron（2μm）laser resection of prostate and bladder neck were established using healthy adult canines.Wound specimens were harvested at 3 days and 1,2,3,4,8 and 12 weeks after surgery respectively.The hispathologic characteristcis were observed by hematoxylin and eosin（HE）staining.Immunohistochemical staining was used to detect TGF-β₁ and CKIP-1 expression at 3,4,8 and 12 weeks.（2）Construction of CKIP-1 over-expressing BPH-1 cells and detection of TGF-β₁ expression by Western Blot.Non-contact co-culture system was used for revealing the effects of CKIP-1level of prostatic epithelial cells on the expression of TGF-β₁ andα-SMA in fibroblasts of prostatic urethral in vitro.Natural BPH-1 cells or CKIP-1over-expressing BPH-1 cells were co-cultured with HFF-1 cells for 72 hours respectively,and then Western Blot was used to detect the expression of TGF-β₁ andα-SMA expression in HFF-1 cells.Results:（1）The urethral wound of the prostate gland is proliferated by the basal cell of the prostate duct or the glandular vesicles under the wound.After the migration and germination after covering the wound,the epithelium was completed,and after the restoration,there were a large number of prostatic epithelium and a small amount of scar formation in the wound,and the bladder neck wound was further differentiated by the urothelium proliferation,migration and covering of the wound of the edge of the urinary pathway.After the repair,there were no epithelial cells in the wound,and scar tissue was found in the wound.;（2）Compared with the wound of 3 weeks after operation,4,8 and 12 weeks of the expressions of TGF-β₁（p<0.01,p<0.05）in prostate epithelial cells and fibroblasts was decreased in canine prostatic urethral wound.Compared with the wound of 3 weeks after operation,4,8 and 12 weeks of the expressions of CKIP-1 in prostate epithelial cells increased（p<0.01）in canine prostatic urethral wound.There was no significant change in the expression of TGF-β₁ of fibroblasts in the wound of bladder neck in 3,4,8 and 12 weeks after operation.Compared with the expression of fibroblast TGF-β₁ in the wound of 4,8 and 12 weeks after operation.4,8 and 12weeks after operation,The expression of fibroblast TGF-β₁ in the urethral wound of prostate was lower than that of fibroblasts in bladder neck wound（p<0.01）.Both the prostate and bladder neck wound fibroblasts were weakly CKIP-1 expressed and there was no significant changes in 3,4,8 and 12 weeks;（3）Compared with the control group,the expression of TGF-β₁ in BPH-1 cells with CKIP-1 expression decreased25%（p<0.05）.The expression of TGF-β₁ andα-SMA in HFF-1 cells of co-culture group decreased by 20%and 30%compared with control group.（p<0.05）,the expression of TGF-β₁ andα-SMA in HFF-1 cells of over-expressing and co-cultured group was reduced by 15%and 10%compared with co-culture group（p<0.05）,respectively.Conclusions:（1）A large number of prostate epithelial cells in the urethra of prostate gland may be one of the causes of the wound scar healing;（2）the prostate epithelial cells may regulate the function and prognosis of fibroblasts in the later stage of repair,which may reduce the expression of TGF-β₁ by raising CKIP-1expression,It inhibited the expression of TGF-β₁ in peripheral fibroblasts and the transformation of fibroblasts into fibroblasts,reduced the excessive proliferation of fibrous cells and the excessive formation of scar.