| 【Objective】1. To establish a simple and feasible, stable, reproducible rat portal veinarterialization model(complete portal vein arterialization and partial portal veinarterialization).2. To explore the effects of accepting different blood supply(venousblood,venous and arterial blood,arterial blood) in portal vein of rats with the liverischemia-reperfusion injury.【Methods】1. Experiment one:Establishment and observation of rat model.1.1Complete portal vein arterialization model: Using the ‘’type allograftmaterials(upper end of the sleeve cuff, the lower end ligation side end exclusion) thensets into the suture to establish the model. Removal of the left kidney, the left renalvein and the portal vein stump were connected by cuff, and the left renal vein and thesuperior mesenteric vein were connected by the cuff.1.2Partial portal vein arterialization model: Using the ‘’type allograftmaterials(on the lower end of the sleeve cuff, side end exclusion) and setting into thesuture cuff to establish the model. Removal of the left kidney, the left renal vein andthe portal vein stump and were connected by cuff, and the left renal vein, the superiormesenteric vein and the superior mesenteric vein were connected by the cuff.1.3Observation:50SD rats were randomly divided into3groups: completeportal vein arterialization group(n=20),partial portal vein arterialization group(n=20)and the sham-operated control(n=10) which only remove the left kidney and one ofthe branches of the free portal vein. Analizing the operative time,blood loss,occlusiontime of portal vein and survival rate.2. Experiment two:Investigation of the effects of accepting different blood supply inportal vein of rats with the liver ischemia-reperfusion injury.108SD rats were randomly divided into3groups: complete portal vein arterialization group(n=36),partial portal vein arterialization group(n=36) and thesham-operated control(n=36).Ensure the blocking time of the first door of the liverwas25min to induce approximately the same degree ischemia-reperfusioninjury.Select6rats radomly,then monitor the portal pressure of invasivly,observe theliver function(peripheral blood ALT,AST), serum NO, plasma ET-1,pathologicalchanges and expression of p38MAPK after2h,6h,1d,3d,7d.3. Statistical analysis: All data were analyzed by SPSS13.0for windows. Allmeasurement data were presented as mean±SD. One-way ANOVA followed by SNKtest was used to compare mutiple groups. The survival time was expressed bycumulative survival curves.Rate was compared by2test or Fisher exact test.Statistical significance was defined as P<0.05.【Results】1. Establishment and observation of rat model:Both models were sucessfully established. The success rate of complete portalvein arterialization model was96.7%（29/30）,operation time was （53±3.95）min,blood loss was (1.1±0.4ml),blocking time was（12.5±1.5）min. The success rateof partial portal vein arterialization model was100%（30/30）,operation time wa（s49.5±3.0）min,blood loss was (0.97±0.33ml),blocking time was （15.2±1.8）min. Thesuccess rate of control without blocking was100%（30/30）,operation time was（23.9±1.85）min,blood loss was(0.25±0.13ml).To compare with the CPVA,blood loss and cumulative survival rate after7d inPPAV had no significant difference（P>0.05）.Operation time was shorter but blockingtime was longer in the later,the difference was significant (P <0.05).2. Effects of accepting different blood supply in portal vein of rats with the liverischemia-reperfusion injury:2.1Serum ALT and AST:After operation,serum ALT and AST of CPAV andPPVA in2h,6h,1d,3d were lower than sham-operated control.ALT in PPVA after2h,6h,1d,3d and AST after2h,6h reduced significantly than CPVA（P<0.05）.2.2Serum NO:Compared with the sham group, the NO of CPAV and PPVA after2h,6h,1d were significantly lower(P<0.05) expecially PPVA continued to reduceuntill3d. Compared with CPVA,NO in PPVA after2h,6h,1d reducedsignificantly(P<0.05). 2.3Plasma ET-1: Compared with the sham group, the ET-1of CPAV and PPVAafter2h,6h were significantly lower(P<0.05) expecially PPVA continued to reduceuntill1d while CPAV after7d was higher(P<0.05). Compared with CPVA, ET-1inPPVA after2h,6h,1d,7d reduced significantly(P<0.05).2.4Liver pathology HE and results：Ischemia-reperfusion injury in control wasmost serious and recover most slowly. we can see a large number of liver cellsdisorganized, edema, vacuolar degeneration, sinusoidal congestion associated with asmall amount of massive neutrophil exudation and Kupffer cell aggregation, hepaticlobule is unclear, and a small amount of liquefaction necrosis. CPVA expressed visiblehepatocytes slight edema, a small amount of liver cells showed vacuolar degeneration,sinusoidal slight congestion, and neutrophil exudation and Kupffer cell aggregation,lobular architecture was unclear. Visible liquefaction necrosis of liver cells flake.Ischemia-reperfusion injury in PPVA group was lightest and recovered mostfastest.The hepatic lobule central vein, portal area structure clear, liver cell cordarranged in neat rows, a small amount of inflammatory cell infiltration; the sinusoidalstructure rules, sinusoidal endothelial cells swellmild congestion. Occasionallyliquefaction necrosis of liver cells flake.2.5Expression of p38in liver:All time pointsin both CPAV and PPVA,p38wereincreased firstly and then decreased expecially after6h expressed most. In sham groupeach time point p38showed a decreasing trend, almost no expression after1d. P38protein expression in three groups had no significant difference.After6h,p38in CPVAand PPVA were higer than control and the former was lower than the later.【Conclusion】1. Establish the model of portal vein arterialization by cuff method using allografta issimple and feasible, reproducible and has high success rate.2. The CPVA group and PPVA group expecially the latter are more resistant toischemia-reperfusion injury compared to the sham-operater control.This may relateto the activated p38MAPK signal pathway resulting in inhibition of theinflammatory response. |
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