The Expression Of Matrix Metalloproteinases And Its Tissue Inhibitors In Bile Duct Malformation And Clinical Significance

Part I The expression of MMP-1, MMP-2,TIMP-1, TGF-β₁ in liver of biliary atresia and its relation with hepatic fibrogenesisObjective: Progressive hepatic fibrosis is an important factor to affect the prognosis of BA after Kasai operation. In order to search for new treatment for BA, the expression ofMMP-1,MMP-2,TIMP-1 , TGF-β₁ in BA liver and their relations with hepaticfibrosis were studied.Methods: There were totally 33 cases in the study. Thirty-three liver specimens were taken from the margin of livers in the operation . The immunohistochemical method(SP) was used to detect the protein of MMP-1,MMP-2,TIMP-1, TGF-β₁ Hepatic fibrosis wasclassified into I to IV grades according to Ohkuma's standards by HE-stains. Twelve liverspecimens of normal infants served as controls. The positive expression was quantitatively analyzed by image analysis technology.Results: The positive expression in BA liver was mainly located at hepatic cytoplasm and membrane. The positive expression was also observed in the bile ductular epithelial cells and Kupffer cells â‘ The increased expression of TGF-β₁,MMP-2,TIMP-1 were parallel to the degree of hepatic fibrosis((r=0.6179, P<0.05; r=0.5063, P<0.05; r=0.5804, P<0.05, respectively) â‘¡ NO parallel relation was observed between positive expression of MMP-1 and the degrees of hepatic fibrosis( r=0.2723, P>0.05). â‘¢ The positive expression of TGF-β₁ was parallel to that of MMP-2 and TIMP-1 in the development of BA liver fibrosis (r=0.9200, P<0.01; r=0.7953, P<0.01). No parallel relation was observed between TGF-β₁ and MMP-1 . The image analysis showed that the expression of TGF-β₁,MMP-2,TIMP-1 were markedly higher in BA livers than in control ( P<0.01 ) ,there was no difference between two groups in the expression of MMP-l(P>0.05).Conclusions: â‘ TGF-β₁,MMP-2 and TIMP-1 are closely related to the development of BA hepatic fibrosis. â‘¡TIMP-1 may be involved in BA hepatic fibrosis by suppressing MMP-1. â‘¢It could become an alternative therapy for BA to decrease the expression of TGF-β₁,MMP-2 and TIMP-1, or improve the expression of MMP-1. Part II Association between the expression of MMP-1 ,MMP-2, TIMP-1,TGF-β₁ and the prognosis in biliary atresiaObjective: The outcome of BA depends on the patient's age at operation, type of BA, severity of hepatic fibrosis and postoperative complications. The histological appearance of liver is progressive fibrogenisis, which is an important factor for prognosis after kasai procedure. In present study, the expression of MMP-1,MMP-2, TIMP-1,TGF-β₁ were observed in different prognosis groups after Kasai procedure .Methods: The clinical data and liver specimens of nineteen BA were collected. Serum value of total bilirubin,direct bilirubin,albumin and GPT before operation were summarized retrospectively. The hepatic specimens were taken from the margin of livers in the operation. Hepatic fibrosis was classified into I to IV grades according to Ohkuma's standards by HE -stains. The immunohistochemichal method (SP) was used to detect the expression of MMP-1,MMP-2,TIMP-1,TGF-β₁. The image analysis technology was used to quantitatively analyze the protein expression. Results: BA children were divided into two groups ( favorable prognosis group and unfavorable prognosis group, briefly called FPG and UFPG) according to their outcome after Kasai procedure. There were seven and twelve patients in FPG and UFPG, respectively .The average age (days) at operation were 58.57±9.32; 122.25±54.67 in FPG and UFPG, respectively. There was obvious difference in age between two groups (p<0.01). The hepatic fibrosis in UFPG was more serious than that in FPG (p<0.05). There was no difference in preoperative serum value of total bilirubin,direct bilirubin, GPT, albumin between two groups(p>0.05). The expressions of TGF-β₁,MMP-2, TIMP-1 were markedly higher in UFPG than in FPG (p<0.01), there was no difference in MMP-1 expression between two groups(p>0.05)Conclusions: â‘ The prognosis of BA is closely related to the age at operation and hepatic fibrosis, not to preoperative serum value of total bilirubin ,direct bilirubin, GPT, albumin. â‘¡TGF-β₁,MMP-2 and TIMP-1 play an important role in the procession of liver damage and fibrosis in BA , their overexpression is related to rapid hepatic fibrosis and poor outcome of BA. The liver transplantation should be early prepared in these patients. Part III The expression of MMP-1,MMP-2 and its tissue inhibitors TIMP-l,TIMP-2 in congenital choledochal cystObjectives: To detect the expression of MMP-1, MMP-2, TIMP-1, TIMP-2 in congenital choledochal cyst(CCC) and its relation with CCC pathogenisis.Methods: The clinical data and specimens of CCC wall and its gallbladders were collected. There were fifty three CCC cases in the study, including twenty-one boys and thirty-two girls aging from four months to seventeen years old (average six point five years old). Twenty-eight normal gallbladders served as controls. The microstructures of the specimens were observed by the optic microscope after HE-stains. The immunohistochemical methods (SP) was used to detect the expression of MMP-1, MMP-2, TIMP-1, and TIMP-2. The quantity was got by image analysis technology. Results: All cases were observed by the optic microscope after HE-stain. Fiber hyperplasia and thick wall were observed in 45 (85%) cases, capillary congestion and inflammatory cell infiltration in 47 (89%) cases, the destroyed mucosa membrane and split muscle fibre in 50 (94%) cases. The lesions in CCC wall were more serious than that in CCC gallbladder and control group. The expression of MMP-1, MMP-2, TIMP-1, and TIMP-2 were located at epithelial cells, matrix cells and inflammatory cells of CCC wall. The various expression can also be found in mucosa membrane cells of CCC gallbladder. Quantitatively, the expression of MMP-1, 2 in CCC wall is markedly higher than that in CCC gallbladder and control group (p<0.01). The expression of TIMP-1,2 were higher in CCC walls and CCC gallbladders than in control group (p<0.01), there was no difference between CCC wall and CCC gallbladder(p>0.05).Conclusions: â‘  The destroyed mucosa membrane and matrix were common in CCC walls, the pathological change was serious in CCC wall than in CCC gallbladder and control group .â‘¡ The expression of MMPs/TIMPs were imbalanced in CCC wall. The overexpression of MMP-1 and 2 may be involved in pathogenisis of CCC.