目的研究肝移植术后胆道铸型(bile cast,BC)的组织结构和溶解特性,探讨其形成机理和溶解治疗的可行性。方法(1)组织化学染色,光镜下观察:将BC制成石蜡切片,分别进行HE染色、胆红素染色、纤维素染色、黏液物质染色和Masson染色后,光镜下观察BC的组织构成。(2)切取小块BC组织,扫描电镜观察超微结构。(3)溶解试验:切取小块BC组织,放入试管中,应用糜蛋白酶、肝素、胰蛋白酶和不同pH值的酸碱溶液,对铸型进行溶解,观察其溶解特性。结果(1)BC大部分成分是胆红素结晶,少部分由胶原纤维组成,其中含有纤维素渗出,夹杂有小血管,有少量的胆管上皮细胞和成纤维细胞,未见坏死的胆管结构。(2)扫描电镜下观察,有不同形态的晶体形成,表面有黏液样分泌物和少许坏死脱落的上皮细胞。(3)BC在碱性溶液中部分溶解,在酸性溶液中不溶解。应用糜蛋白酶、肝素、胰蛋白酶对Bc没有溶解作用。结论(1)BC是以胶原纤维为支架,充填大量的胆红素结晶成分;(2)纤维蛋白、成纤维细胞和血管的出现,提示铸型的形成和发展可能是胆管上皮损伤、渗出、机化和吸收的动态变化过程;(3)结晶体形成的机理不清,可能与胆汁性状发生改变,使胆红素等物质易于析出形成结晶;(4)BC在酸性环境下不溶解,碱性环境下有一定的溶解性,糜蛋白酶和胰蛋白酶对其不能溶解。
Objective To investigate the histological components and dissolvability of the bile-cast, and to study the pathological course of bile cast formation. Methods HE staining, bilirubin staining (Gmelin reaction) , Masson's staining, alcian blue staining and fibrin staining (weigert's) were performed on the formalin-fixed paraffin-embedded section of the bile cast. Uhrastructure was examined under the scanning electron microscope. Dissolvability test was also conducted using chymotrypsin, heparin, trypase solution, HCI and NaOH solution to dissolve the bile-cast. Results The major components of the bile-cast were bilirubin crystals and collagen fibers. Between the mass of collagen fibers there was certain blood vessel structure. Necrosis bile duct structure was not found in the cast. Under the scanning electron microscope, four kinds of crystal morphouses were viewed. There were some mucoid mass and necrosis defluvium epithelial cells in the bile cast. Dissolvability test showed that the bile cast could be partial dissolved in NaOH solution ( pH = 12.5 ). No dissolution was found in HCl solution ( pH = 5.0 ) , chymotrypsin solution, heparin and trypase solution. Conclusions Collagen fibers work as framework in the bile cast with bilirubin crystal filling between the framework. The emergence of fibroblast and blood vessels indicated the formation of bile cast might be the course of exudation and organization due to bile duct epithelium damage. Bile cast could be partially disssolved in alky solution, but could not be dissolved in acid solution, or in chymotrypsin, heparin and trypase solutions.