缓释碱性成纤维细胞生长因子预防胆管损伤修复术后胆管瘢痕形成的实验研究  

The prevention of bile duct scar formation after bile duct injury repair by using sustained-release basic fibroblast growth factor

作　　者：陶亮[1] 李强[1] 任昊桢[1] 戴建武[2] 丁义涛[1] 

机构地区：[1]南京大学医学院附属鼓楼医院肝胆外科,南京210008 [2]中国科学院遗传与发育生物学研究所,北京100109

出　　处：《南京大学学报（自然科学版）》2012年第6期797-803,共7页Journal of Nanjing University（Natural Science）

基　　金：南京市医学科技发展一般课题基金(YKK11087)

摘　　要：探讨缓释碱性成纤维细胞生长因子(basic fibroblast growth factor,bFGF)对胆管损伤修复术后胆管瘢痕形成的预防作用.通过分组实验,制备猪胆管部分缺损模型,运用加载缓释bFGF胶原膜及空白胶原膜修复缺损.比较术后1、2、3个月的肝功能变化并用病理切片和免疫组化染色比较胆管愈合、胶原增生及肌成纤维母细胞(myofibroblast,MFB)的表达情况.术后6个月胆道造影观察狭窄形成情况.结果显示:经缓释bFGF治疗后,胆管未出现瘢痕性狭窄.与空白胶原膜组相比,胆汁淤积酶学指标中GGT和TBA显著降低,免疫组化提示MFB的表达显著降低,胶原排列及结构更接近于正常胆管.证明运用缓释bFGF胶原膜治疗胆管缺损可通过加速胆管愈合及下调MFB的表达有效的预防胆管损伤修复术后胆管瘢痕的形成.The over-healing is a major problem in the process of bile duct injury repair. It will cause malignant outcomes because of stricture by scar formation. It is necessary to promote rapid-healing of the injury to prevent scar formation. In our previous work, we produced a sustained release basic fibroblast growth factor（bFGF）. After adding to collagen membrane it could not only retain the activity of growth factor but also obtain the ability of sustained and site-targeting. In our study,we used this material to repair a 2.0 ×0.5cm elliptical defect in the bile duct of pigs,and blank collagen membranes with same size were used as control. Before being sacrificed,the alanine aminotransferase （ALT）, γ-glutamyltransferase（GGT）, direct bilirubin（DBil）, and total bile acid （TBA） in both groups were detected and compared. After being sacrificed,the H＆E staining and Masson＇s trichrome staining of the neo bile duct and the immunohistochemical staining of a SMA in both groups was observed and counted, the anti-a SMA antibody was used to mark the myofibroblast （MFB）which is an important factor in the process of scar formation. The cholangiography was performed under fluoroscopic guidance by cholecystostomy to evaluate if there was any evidence of stricture formation at 6 months. Hg〉.E staining showed the faster healing rate of neo bile duct and less scar formation after treated by sustained-release bFGF,and the thickness of neo bile duct was similar with normal bile duct at 3 weeks. Compared with control group, the levels of GGT and TBA were significant low at all time points;immunohistochemical staining showed the expression of MFB was also significant low at all time points. The structure of collagen fibers was similar with the normal bile duct as showed by Masson＇s trichrome staining. It is proved that using a sustained-release bFGF could effectively prevent postoperative bile duct scar formation by accelerating the healing rate of bile duct and down-regulating the expression of MFB.

关 键 词：肝外胆管 组织工程 肌成纤维母细胞 碱性成纤维细胞生长因子 

分 类 号：Q786[生物学—分子生物学]