Mechanism Of The Biliary Scar And Effects Of Human Caul Fat, Bile And Triptergium Glycosides On The Biliary Scar

Biliary tract scar stricture is the complicated and intractable difficult problem of biliary tract surgery. It is very important significance that the mechanism, prevention and cure of biliary tract scar stricture is deeply studied. The study adopted the methods which was combined cilinic with basic, and explored the morphologic character and mechanism of the biliary tract scar by the numbers, and investigated the mechanism of biliary tract scar and the effects of human caul fat, bile and triperygium wilfordii polyglycosidium on the biliary tract scar. 14 cases of biliary tract scar tissues of patients from the operation and 10 cases of normal biliary tract tissues were collected. The microstructure and ultrastructure of tissues in both groups were studied with light microscope under HE, Masson, Von-Gison's staining and Scanning electron microscope and Transmission electron microscope respective. Intensity of expression and location of CD68, TGF-β₁, a-SMA , c-myc, c-fos, Bcl-2, bax, Survivin, bFGF, VEGF, CD1a and MVD were studied with immunohistochemical and image analysis system. Expression of Bcl-2 mRNA and bax mRNA were studied with semiquantitative RT-PCR in both groups. Content of TNF-a, IFN-γ, IL-2, IL-4, IL-10 were measured with flow cytometry in 14 normal human caul adipose extract and 10 normal human blood. Content of TGF-β₁ were measured with enzyme linked immunosorbent assay in the normal human caul adipose extract and human blood. A model of human caul fat and human biliary tract scars were established with implanting the fat and the scars to subcutaneous of the nude mice by means of hypodermic or carving and suturing the skin of nude mice. The nude mice were sacrificed after 30 days. The volume of the graft in both groups was measured. The microstructure and ultrastructure of tissues in both groups were studied with light microscope under HE and Von-Gison's staining, electron microscope respective. The effects of human adipose tissue extract, bile and triperygium wilfordii polyglycosidium on the fibroblast were studied by investigating the morphologic character with invert microscope, light microscope under HE staining, electron microscope. The growth of fibroblast was studied by MTT. The cell cycle and apoptosis of fibroblast was studied by flow cytometry. The experimental result showed: The morphologic character of the biliary tract scar tissue include the lasting existence of chronic inflammation, overdisposition of collageous fiber, existence of MFB. Macrophage which deposited the biliary tract wall, effected the vasxularization and fibrosis of biliary tract tissue by liberating much media. The expression of CD1a was positive on the epithelial cell of skin, and was negative on the normal biliary tract and biliary scar. The result showed that dendritic cell may be not concerned with forming the biliary scar. TGFβ1, bFGF and VEGF overly expressed in the the biliary tract scar tissues, promoted the growth of FB, changed FB to MFB, promoted the synthesis of collagen and other stroma and inhibited their degradation, resulted in forming neogenetic blood vessel at the same time. MVD which increased in the biliary tract scar promoted to form scar. MFB(a-SMA) is the princeps factor of the biliary tract scar contraction and the lumen stricture. C-myc and c-fos were actived and obviously expressed in the extrahepatic and intrahepatic biliary tract scar tissues, resulted in scar hyperplasia by effecting the differetiation and proliferation of FB, the synthesis and degradation of collagen,and regulated the cell factors. The first study showed that the expession of survivin was made obviously it up-regulation, inhibited the apoptosis of biliary tract cell to result in overly hyperplasia of biliary tract scar. Bcl-2 and bax were positive expession in the normal bile duct and biliary tract scar. The Bcl-2/bax ratio in the biliary tract scar was significantly highly than in the normal bile duct. The rise of Bcl-2/bax ratio promoted to heavy forming heterodimers Bcl-2/bax, inhibited to forming homotype dimer bax:bax, inbibited the effect that bax promotes the cell apoptosis to result in hyperplasia of scar. The expession of TNF-a,IL-2,IL-4,IL-10 and TGFβ1 in the human caul adipose extract was signifisantly higher than them in the human blood. Adipocytokine may regult the formation of biliary tract scar which is both positivty and negativity. The effect of regulting the formation of biliary tract scar is bothway. The adipose extract inhibited the growth of FB and the heal of biliary tract vulnus. The nude mice homogeneity veriant model which was made by transplanting human biliary tract scar and human caul fat was successfully established by means of carving and suturing the skin of nude mice. The volume of the biliary tract scar graft significantly enlarged and the volume of the adipose graft minified, the morphologic character of graft is invariable after the 30 days. The nude mice homogeneity veriant model which was made by transplanting to combine human biliary tract scar with human caul fat was successfully established. The volume of the biliary tract scar graft was not obvious variety and the volum of the fat graft minfied, but the difference was not signifcant after 30 days. The fat tissue can very well syncretize with the biliay scar tissue. The fat cell and lipid droplet directly immerged in the collagen fibor of the biliary tract scar tissue,especially immerged in the cytoplasm or between FB and collgen fibor. The phenomenon showed that the fat tissue played a role not only by autocrine and paracrine but also by directly immerging in the scar tissue. The fat tissue can inhibit the growth of the biliary tract scar. The bile promoted the growth of FB and promoted to change FB to MFB, at the same time inhibited the apoptosis of FB. The triperygium wilfordii polyglycosidium inhibited the growth of incubatory FB in vitro, and promoted its apoptosis.