小鼠胆管狭窄模型的建立及其缺损修复

Establishment of a bile duct stricture model for bile duct injury and repair in mice

目的通过制造小鼠胆管远端狭窄而近端扩张,建立一种新的胆管缺损与修复研究模型。方法首先构建胆管狭窄模型小鼠并与假手术和胆管完全梗阻(BDL)小鼠比较,检测体重、胆道和肝功变化情况;然后进行胆管缺损与修补,并检测血清生化和组织病理学变化。结果建模术后14天,BDL组小鼠体质量显著低于假手术组(P<0.05),而胆管狭窄组无显著下降;与假手术组相比,胆管狭窄组与BDL组的胆管与胆囊均显著扩张,但二者的直径之和胆管狭窄组显著小于BDL组(P<0.05);吲哚菁绿荧光成像实验证实胆管狭窄组小鼠胆道可通畅引流胆汁;胆管狭窄组小鼠的血清丙氨酸氨基转移酶(ALT)、天门冬氨酸氨基转移酶(AST)和总胆红素水平略高于假手术组(均P<0.05),但显著低于BDL组(P<0.05)。胆管狭窄组小鼠胆管经缺口损伤与修补术后30天,胆管外观修复良好,胆道仍然通畅;HE染色见补片处胆管上皮排列整齐;免疫组化证实绿色荧光蛋白、CK19染色阳性。结论成功构建小鼠胆道缺损与修复模型,可为胆管损伤修复机制的研究和组织工程胆管的评价提供一种新的模型。

Objective To establish a new bile duct injury and repair model in mice by generating bile duct distal stricture and proximal dilatation.Methods The mice were randomly divided into sham operation group,bile duct stricture(BDS)group and bile duct ligation(BDL)group.The dilated bile duct of BDS mice was injured and then repaired 14 days after the modeling operation.Biochemical markers were detected and histopathological changes were analyzed.Results 14 days after the establishment of the model,the body mass in BDL group was significantly lower than that of the sham group(P<0.05),while the body mass in BDS group was similar to sham group.Compared with the sham group,the bile duct and gallbladder of the BDS group and BDL group were both prominently dilated,but the sum of the diameters of bile duct and gallbladder in BDS group was significantly smaller than that in the BDL group(P<0.05).Indocyanine green fluorescence imaging confirmed that biliary tract of BDS group could still drain bile.Serum ALT,AST and TBil levels in the BDS group were slightly higher than those in the sham group(all P<0.05),but significantly lower than those in the BDL group(P<0.05).Bile ducts of BDS mice were injured by notching and repaired with bile duct path.30 days after the repairing,HE staining showed that the bile duct epithelium around the patch was arranged in orderliness.Immunohistochemistry confirmed the positive staining of green fluorescent protein(EGFP)and CK19 in those groups.Conclusion This model of bile duct injury and repair in mice can provide a new model for the study of the mechanism of bile duct injury and repair and the evaluation of tissue engineering bile duct.