| Ischemia-reperfusion injury (ischemia reperfusion injury, IRI) is common in asurgical operation; the many kinds of factors involved the path physiological process. IRIcan damage the ultra structure of because a series of inflammation causes the cell toedema, and cause organ function damage and even death. Liver IRI is more than amolecular; many factors are involved in pathological process, common in livertransplantation, severe liver injury treatment, such as liver resection of clinical liversurgical treatment process. In view of its specific pathogenesis is a relatively complex,and pathologic outcome for liver more cell apoptosis, necrosis, the serious influenceclinical outcomes, so its pathogenesis has become a hot research topic today. Thepathogenesis of major relevant generally summarized by the following five:1.calciumoverload;2. oxygen free radicals (ROS);3. Cell factors;4. Cell apoptosis, necrosis;5.Europhiles and related factors. Now many people think that Wnt/β-catenin signalpathway in the liver of occurrence and development, nutrition and metabolism, liver cellregeneration and cancer happening all play an important regulatory role. β-catenin willlead to liver damage to all sorts of wounding stimulates the sensitivity of the increase,but the mechanism of liver in ischemia reperfusion injury is not clear.β-catenin is a relative molecular weight for92000cells glycoprotein, gene in thechromosomes of the positioning is3p21, normally located mainly in the cell membranes,and participate in the epithelial sticky protein (E-Cad) mediated cell adhesion, but thesignal transduction pathway in a more important role. The cytoplasm of β-catenin andshaft protein, colonic adenoma sample polyps (adenomatous polyposis coli, APC)protein, casein kinase1(casein kinase1, CK1), glycogen and enzyme kinase3β (GSK3β)might be constitute a complex, including shaft protein and APC protein as a scaffoldprotein, and in the β-catenin CK1and GSK3b phosphorylation on it, finally β-catenin inprotease under the function of be broken down. When β-catenin lost phosphorylation willfall out of from the complex, to the cytoplasm and the nucleus accumulation, and TCF/LEF union, start and cell proliferation or antiapoptotic related gene transcription,this is typical Wnt/β-catenin signal pathways. Wnt signal system has much effectpleiotropics, the study found that the Wnt/β-catenin axis in the liver of occurrence anddevelopment, nutrition and metabolism, liver cell regeneration and cancerous tumors etcplays an important role[21,22].Although these studies support β-catenin biologicalactivities with TCF/LEF mainly by relevant Wnt signaling to mediated, recent studiessuggest that β-catenin can also play with the TCF-independent transcription auxiliaryactivator effect[3]. In normal physiological condition, the β-catenin biological activitiesmainly by and lymphatic enhancement factor/T cells factor (TCF/LEF) of the Wnt signaltransduction mediated by, and in adverse pathology condition (hypoxia or ischemiareperfusion), β-catenin may play has nothing to do with the TCF transcription auxiliaryactivation[5], help liver cell adaptation environment and enhance the survival ability.Therefore, the study of β-catenin in hepatic ischemia/reperfusion and its regulatorymechanism for a better understanding of ischemia/reperfusion injury inflammationmechanism has important scientific significance.In this study, first constructing liver cell specificity knockout β-catenin mice tocut off the tail, DNA genome extraction, PCR, gelatin electrophoresis, DNA sequencingetc. Result showed that, liver cells specific knockout β-catenin mice construct success.After and related purpose strip contrast, we set of liver cells specific knockout β-cateninJackson laboratory mice with the purpose of provided bands are identical with each other.Through the liver cells-specific knockout mice β-catenin establish partial liverischemia-reperfusion model, detection β-catenin gene knock out mice with a controlgroup of liver ischemia-reperfusion injury serum alanine aminotransferase (ALT),aspirate aminotransferase (AST) and its histological change, explore cause liverdamage,discusses the role of β-catenin gene in hepatic ischemia-reperfusion. Part IConstruction of β-catenin-deficient Miceβ-catenin is an important member of Wnt signal pathway in many physiologicalpathology participates in the process the adjustment. In recent years, research shows thatβ-catenin in liver cell growth, development, differentiation, stress as process has played animportant role. Then select β-catenin as the experiments of genes, we applied Cre-loxsystem to construction liver cell specificity knockout β-catenin genetic mice model. FromJoint Ventures Sipper BK animal laboratory of purchase C57BL/6transgenic mice,4-6weeks, B6. Cg-Tg (MX1-cre) Cgn/J mice and β-catenin loxP/LoxP (Ctnnb1fl/flmice,Jackson laboratory) hybrid, get Mx-cre×Ctnnb1fl/flmice genomic DNA confirm whetherfor Mx-cre×Ctnnb1fl/flmice, were three times to Ctnnb1fl/flmice and Mx-cre×Ctnnb1fl/flmice intra-abdominal injection poly (I: C),2days injection once, the6thdays to give thelast an injection, total about250μ g measure (12.5u g/kg), and then will Mx-cre×ctnnb1fl/flmice cut tail again PCR, gelatin electrophoresis, confirm the successful constructionβ-catenin defect mice. Experimental Study on the Protect Effect of β-catenin in MouseLiver Ischemia Reperfusion InjuryObjective: In addition to the β-catenin gene in the mouse ischemia reperfusion injury inthe mouse model to investigate the role by constructing specific knock. Methods: usingCre-lox system building liver cells specific gene knock out β-catenin mouse models,60min half liver blocked blood flow manufacturing liver ischemia-reperfusion mouse models,respectively for β-catenin knockout group and control group, and after6h reperfusion,cutting killed mice, the inferior vena cava take blood, centrifugal, and the measured serum alanine aminotransferase (ALT), aspertate aminotransferase (AST); Cut off the left lobe ofliver,10%formalin fixed. Liver tissue after more than liquid nitrogen freezing kept in-80℃, and then sent to theChangzheng hospital hepatic pathology, the first hospitalorganization HE dyeing and immunohistochemical (IHC) test, observation liver pathology;Remaining liver tissue real-time PCR detection quantitative fluorescence line (RT-PCR),observe the inflammatory factor expression. Results: the β-catenin knockout groupserological shows that (ALT AST) were higher than those in the control group (p <0.01),compared with the control, β-catenin knockout group of liver cells of expression ininflammatory factor protein abundance to significantly higher than the control group (p <0.01); Compared with the control, β-catenin knockout group of paraffin section ofhematoxylin-eosin (HE) stain reveals the necrosis of liver cells, sinusoids congestion andinflammatory cell significantly heavier than the control group. Conclusion: β-catenin geneon liver ischemia-reperfusion-induced liver injury play a protective role. |
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