| OBJECTIVE1. Design a steady,credible rat model of schistosomal portal hypertension. The model accord with the nature course of schistosomiasis. To observe the change of liver criihosis and portal hypertension.2. To study the relation of gut barrier and endotoxemia. To explore the track of endotoxin through gut barrier.3. To study the changes of expression of Toll-like receptor 4 and myeloid differentiation protein-2 of mesenteric vein in rats with Schistosomal portal hypertension. To explore the role of intestinal endotoxemia in mesenteric vein vasculopahty.4. To study the changes of expression of interleukin 1 receptor-associated kinase 4 of mesenteric vein in mice with Schistosomal portal hypertension.METHODS1. Infected mice percutaneously with cercariae of schistosomiasis japonica. Subdividing mice into 6 week,12 week and 20 week groups. HE stain,Masson trichrome stain and transmission electron microscope examination was used to observe pathologic pattern of liver and colon. The diameter of portal vein and portal venous pressure was used to observe the change of flow dynamics.2. To observe the change of gut barrier function by FITC-D. Endotoxin was measured by MT-1 reagent box. Liver tissue was collected for germiculture. Immunohistochemistry and quantitative analysis of Western blot were applied to detect the protein expression of occluding. TUNEL methods was applied to detect the apoptosis of colon mucous membrane cell.3. RT-PCR were applied to detect the mRNA expression of TLR-4/MD-2 and TNF-αin the mesenteric vein. Analyzed the pertinency between expression of TLR-4 and endotoxemia.4. RT-PCR and Western blot were applied to detect the protein and mRNA expression of IRAK-4 in the mesenteric vein. Analyzed the pertinency between expression of IRAK-4 and TNF-α.RESULTS1. The infected rate of model mice is 100%, mortality rate is 36%. At the 12th week after infection, the liver fibrosis was occurred; at the 20th week after infection, the liver cirrhosis was occurred. Portal venous pressure was gradually raised at 12th week group, and keeping a high level at 20th week group.2. The concentration of FITC-D and in 12th week and 20th week group is significantly stronger than normal group (p<0.01), this trend was positively correlated to increased the concentration of endotoxin(r=0.543,p<0.01). The ratio of bacteria translocation in 12th week and 20th week group is also significantly higher than normal group (p<0.01). The expression of occludin in 12th week and 20th week group is significantly lower than normal group; The apoptosis index of 12th week and 20th week group is significantly higher than normal group.3. The expression of TLR-4,MD-2 and TNF-αmRNA in 12th week and 20th week group is significantly stronger than normal group (p<0.01). The expression of TLR-4/MD-2 mRNA was positively correlated to increased of the concentration of endotoxin(r1=0.548, p<0.01;r2=0.534, p<0.01). The expression of TNF-αmRNA in 12th week has no significantly difference(p>0.05).4.The expression of IRAK-4 protein and mRNA in 12th week and 20th week group is significantly stronger than normal group (p<0.01), comparing 12th week and 20th week group, there has no significantly difference(p>0.05). The expression of IRAK-4 mRNA was positively correlated to increased of the expression of TNF-αmRNA(r=5.56, p<0.01). CONCLUSIONThe mice model establish by infecting percutaneously with cercariae of schistosomiasis japonica has a nature course of schistosomiasis. It has a significantly change of portal flow dynamics. It is a appropriate model to study schistosomal cirrhosis and portal hypertension. The development of portal hypertension is accompanied by the damaged of gut barrier and portal endotoxemia. The raise of intestinal permeability is the main reason of intestinal endotoxemia. LPS can upregulate the expression of its signaling receptor :TLR-4. Through the TLR-4 signaling pathways, LPS can induce IRAK-4 is the key regulator in the TLR-4 signaling pathway, it play an important roll in portal hypertension vasculopahty. |
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