肝细胞生长因子促进膀胱平滑肌创伤愈合的研究

HGFs accelerated smooth muscle wound healing and impaired local scar response in rabbits bladder

目的 用单纯膀胱肌肉损伤模型研究肝细胞生长因子 (HGFs)质粒DNA(pUDKH)在膀胱平滑肌创伤愈合中的作用。方法 用 2 4～ 3 0周新西兰兔制备单纯膀胱前壁肌层缺损创面 3个 ,面积均为 1cm2 大小。分别涂覆单纯溶媒 (A组 ) ,15 μg空载体质粒DNA(pUDK ,B组 )和 15 μgpUDKH(C组 )溶液 ,只涂 1次 ,涂覆次序轮替 ,共 2 0组。术后进行系列创面愈合速度检查 ,影像检查和组织学检查 ,比较肥大指数 (HI)。结果 术后 2 0只兔均存活至实验结束。术后各组平滑肌缺损创面均缩少 ,A ,B和C组创面面积 ,7?d时分别为 ( 4 3 .9± 9.8)mm2 ,( 4 3 .0± 11.1)mm2 和( 2 7.7± 7.5 )mm2 ,14d时分别为 ( 2 4.1± 9.1)mm2 ,( 2 2 .4± 7.4)mm2 和 ( 4 .1± 4.0 )mm2 ,C组与A、B组相比差异显著。膀胱影像检查 ,各组膀胱膀胱形态基本正常。术后组织学显示 ,A ,B组显示上皮层正常 ,粘膜下纤维组织增生加厚而肌层较薄 ;C组显示与自体膀胱具相似的组织学结构。术后 3 0d时 ,A ,B和C组的HI分别为 ( 1.76± 0 .17) ,( 1.83± 0 .18)和 ( 1.11± 0 .18) (P <0 .0 1,CvsAandB)。结论 实验组兔膀胱创面愈合速度快 ,对照组创面膀胱平滑肌再生速度慢 ,再生不完全 ,粘膜下纤维瘢痕增生。提示HGFs能够促进膀胱平滑肌创伤愈合 。

Objective To study the role of the reconstructed plasmid (pUDKH) of hepatocyte growth factor (HGFs) in smooth muscle wound healing and local scar response in rabbits bladder.Methods The pUDKH was obtained from a eukaryotic expression vector pUDK inserted with a cDNA clone for human HGF isolated from human placental cDNA library.A total of 20 New Zealand rabbits underwent a cyst-muscle-ectomy on the anterior bladder wall to make three wounds about 1 cm2.Menstruum only (group A),15 μg pUDK (group B) and 15 μg pUDKH (group C) was embrocated respectectly and alternately in every bladder.Preoperative and postoperative gross,histologic and hypertrophic index (HI) analyses of the wound healing studies were performed serially.Results All 20 rabbits survived until the end of the experiment.Postoperative bladder wound area in three groups (A,B and C in turns) were (43.9±9.8) mm2,(43.0±11.1) mm2 and (27.7±7.5) mm2 respectly on day 7 and (24.1±9.1) mm2,(22.4±7.4) mm2 and (4.1±4.0) mm2 respectively on day 14.The wound area reduction in group C was significantly different that that in group A or group B (P<0.01).Postoperative bladder wound hypertrophic indexes in groups A,B and C were 1.76±0.17,1.83±0.18 and 1.11±0.18 (P<0.01,C vs A and B) respectively on day 30.Grossically and radiographically,all bladder maintained normal shape.Histologically,the menstruum only and pUDK treated bladders presented a pattern of normal urothelial cells with a thickened fiberotic submucosa and a thin layer of muscle fibers.The retrieved pUDKH treated bladders showed a normal cellular organization consisting of a trilayer of urothelium,submucosa,and muscle.HI in group C was lower than that in groups A and B group (P<0.01,C vs A and B).Conclusion The application of pUDKH in bladder wounds could improve the smooth regeneration and prevent the scar formation.