| PARTI Establishment of Rats Model about associating River partition and Portal Vein ligation for staged hepatectomy(ALPPS)Objetive The aim of the study is to establish a rat model of associating liver partition and portal vein ligation for staged hepatectomy(ALPPS)Method Twenty healthy male SD rats,weight 200-240g.The rats were anesthetized by injecting 10% chloral hydrate, the abdomen was opened through a longitudinal incision after anesthesia, the abdomen retracts the incision and holds it, periostical ligament and each liver lobe were freed ,we opened corresponding glisson sheate, ligatured the right lobe portal vein and portal vein left branch (including left lateral lobe and left middle lobe) with 4-0 silk,respectively .Then we removed the caudate lobe(It is difficult to ligature the portal vein of caudate lobe).At the time,ischemia zone appeared between the left and the right middle lobe after ligation.We transected the liver parenchyma along with the ischemic zone . During the course of the operation, oppression or suture method were used to stop bleeding. Warm saline was used to rinse the abdominal cavity after check no abnormal, abdominal incision was closed by silk continuous suture. The establishment of the whole model is divided into two stages of training, the stability and repeatability of the model were studied by recording and comparing the success rate of surgery, operation time, surgical survival rate. Seven days later, the surviving rats and 5 normal rats were sacrificed and the corresponding liver lobes were removed. The quality of the right middle lobe of the liver was weighed with an electronic scale . The data collected were analyzed statistically.Results:1. The operation time and the success rate of the surgery were compared between the two stages: the portal vein ligation time, the liver parenchyma transected time and the overall operation time of the second stage were shorter than the first stage (P<0.05). The first stage only 12 rats survived,the survival rate was 60%。The second stage, liver anatomy of rat was studied well and the operation was significantly skillful while the successful rate of operation was 100% which compared with the first stage increased significantly, the difference was statistically significant (P<0.05).2.The two stages postoperative complication rate and survival rate comparison: During the first stage of surgery,complications occurred in 13 of the 20 rats,8 rats were died in the operation, other 5 rats gained complications after operation including 2 rats with postoperative bleeding,2 rats with massive ascites, 1 rat with infection caused by biliary fistula. During the second stage, complications occurred in 1 of the 20 rats. Compared with the first stage, it was significantly reduced, the difference was statistically significant (P<0.05). After training and learning experience of the first stage, during the Second stage, postoperative survival rate of rats was 100%. During the first stage ,of all 20 rats ,8 died in the operation and there were 5 died in the remaining 12 rats,postoperative survival rate was 58.3%, significantly lower than the second stage(P<0.05).3. Seven days after the operation, all the rats were sacrificed. The right middle lobe mass was increased from1.48g±0.10g to2.61g±0.19g, the right middle lobe was significantly increased.Conclusion: we successfully established a stable and repeatable rats model about associating liver partition and portal vein ligation for staged hepatectomy(ALPPS).In this model liver regeneration was very obvious and can be used for liver regeneration related research.PART2 Evaluation of Liver regeneration after ALPPS and its relationship with Wnt2 expression in RatsObjective: To observe the changes of liver function after ALPPS, to evaluate the regeneration of remnant liver, and to explore the relationship between remnant liver regeneration and Wnt2 protein in rats.Methods Sixty healthy male SD rats(200g-240g) were randomly divided three groups:ALPPS group, portal vein ligation (PVL) group ,Sham group. Sham group only freed vein branches without ligation and closed its abdomen. PVL group consisted of the right lobe portal vein and portal vein left branch (including left lateral lobe and left middle lobe) ligation with 4-0 silk and caudate lobe hepatectomy,the right middle lobe was preserved to regenerate. ALPPS group was treated in the same way, but added parenchymal partition (middle lobe). Five rats were sacrificed on day 1,2,4 and 7 in each group after operation to observe the weight of the right middle lobe, respectively.The venous blood were collected for testing alanine aminotransferase(ALT), aspartate aminotransferase(AST),serum albumin(ALB), total bilirubin(TBIL). Histopathologic changes of left middle lobe were observed . The morphological changed of the right middle lobe liver lobes were observed by microscope. The expression of Wnt2 and Ki-67 of the right middle lobe liver lobes were studied with immunohistochemistry. The results were analyzed by medical statistics methods.Results1.The hepatic regeneration rate(HRR) for both ALPPS group and PVL group were obviously higher than the Sham group at all points(allP<0.05). Compared to the PVL group , the HRR of the right middle lobe in ALPPS group was obviously higher on day 4 and 7 [(155.96±24.39)% vs(118.15±9.77 )%,P<0.05; (174.86±8.99)% vs( 133.55± 16.48)%, P<0.05].2.0n day 1 and 2,the serum ALT and AST levels in two groups were obviously increased compared with Sham group.On dayl, the serum AST and ALT levels in ALPPS group were significantly higher than those in PVL group[(2033.40±358.88)U/L vs ( 1343.40±230.75)U/L,P<0.05; (982.60±165.63) U/Lvs(410.80±130.08) U/L,allP<0.05]. There were no significant differences at other time points.Significant lower in the serum ALB concentration were observed in both ALPPS and PVL groups compared with Sham group at any time(allP<0.05).On day land 2, the serum ALB in ALPPS group was obviously lower than that in PVL group, respectively(allP<0.05).TBIL levels had no significantly different at all points.3.More focal necrosis of liver were observed in ALPPS group than those in PVLgroup on day 1.4.Immunohistochemistry results showed that the Ki-67 positive index in the right middle lobe was higher in ALPPS group than PVL group on day 2 and day 4[(85.36±9.13) %vs (61.84±10.14)% ,P<0.05;( 43.40±7.89)%vs (29.06±6.38)%,P<0.05].5.Compared with the Sham group, the Ki-67 after surgery began to increase , it reached the maximum at the second day, then decreased gradually, there were a few positive cells at the seventh day and no statistical significance;6. The positive expression of Wnt2 was mainly around the central vein after ALPPS,it may be associated with the generation of liver stem cells. Compared with Sham group,Wnt2 in remnant liver began to rise after ALPPS, reached the highest at the second day,and then decreased gradually, only a small amount of expression at the seventh day and there was no statistical significance (P>0.05).7.The expression of Wnt2 and Ki-67 had a positive collection after ALPPS.Conclusions1.Both ALPPS and PVL could induce hepatic regeneration of future remnant liver compared with Sham group. However ,ALPPS induced a greater response.2.The expression of Wnt2 have played a pivotal role in liver regeneration and liver stem cell after ALPPS. |
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