| Objective: Exploring the reasons and mechanisms of the differences in the volume of dual grafts after dual-graft liver transplantation in rats.Methods:(1)Establishing a rat dual-donor liver transplantation model: 96 SPF-grade SD male rats,8-10 weeks old,weight(270g-300g),randomly divided into transplantation group(Transplantation group,n=72)and sham operation group(Control group)(Sham group,n=24),in the transplantation group,2 randomly as donors and 1 as recipient.Improved the model of dual-grafts living donor liver transplantation in rat by increasing the weight of SD rats,obtaining the appropriate bile duct length of the donor,and trimming the Y-shaped iliac vessels.(2)A mass of 250-270 g rats with dual grafts liver transplantation had an optimum anesthetic dose of 1ml /100 g,a mass of 200g-250 g was 0.8ml /100 g,a mass of 160-200 g is 0.3ml /100g(3)Observed continuously and dynamically the volume changes of dual grafts after transplantation in rats by magnetic resonance imaging(MRI).(4)Observed continuously the portal vein blood flow(ml/s),portal vein diameter(mm)and portal vein velocity(cm/s)of dual grafts by ultrasound image.(5)Detected the blood biochemical indexes at different time points in the dual-grafts living donor liver transplantation rats after transplantation;(6)Measured the volume and quality of dual grafts from a morphological point of view,and detected the growth of the dual grafts.Growth status of bilateral grafts at different time points after transplantation.(7)HE staining method to detect the growth of dual grafts at different time points after transplantation(8)RT-PCR,western-blot assay to detect the expression of apoptotic proteins such as caspase-3 at different time points after transplantation.Results:(1)Establishing a rat dual-graft liver transplantation model: 96 SPF-grade SD male rats,8-10 weeks old,weight(270g-300g),randomly divided into transplantation group(Transplantation group,n=72)and sham operation group(Control group)(Sham group,n=24),in the transplantation group,2 randomly as donors and 1 as recipient.Through the improvement of operation technology,the survival rate of rats undergoing dual-grafts living donor liver transplantation is significantly increased,the 7-day survival rate is increased from 58.3% to 87.5%,the 30-day survival rate is 68.8%,and complications are reduced.(2)It was found that the volume of the right graft gradually increased,and the left graft had no obvious changes in the early stage and atrophy in the later stage.(3)The portal vein blood flow of the left graft gradually decreased at 1 day,3 days,7 days,and 30 days,and the portal vein blood flow of the right graft slightly decreased from 1 day,3 days,and 7 days after transplantation,but increased significantly at 30 days.(4)Liver function indicators such as AST and ALT showed an increase in the early stage and then decreased during the 30-day observation period.Process.(5)To the 30-day observation period,the total volume and quality of the graft increased significantly compared with before transplantation,and reached the level of the sham operation group,and the volume and quality of the right graft increased more than the left graft(P<0.05).(6)Hepatocytes are severely vacuolated,and there is more inflammatory cell infiltration in the bile duct area in the left graft,the right graft had more obvious proliferation of hepatocytes and bile duct cells,and grows better.(7)PCR and western blot results showed that apoptosis reached the peak 3 days after transplantation and then decreased,and the apoptosis level of the right graft was lower than that of the left,and some apoptotic indicators later(30 days)showed a rise again on the left graft.Conclusions:(1)Techniques such as increasing the weight of SD rats,obtaining the appropriate bile duct length of the donor,and trimming the Y-shaped iliac vessels can significantly improve the survival rate of rats with dual-grafts living donor liver transplantation,making the model more stable.(2)The weight of dual-graft liver transplanted rats is related to anesthesia dose;(3)The growth trend of the right graft is generally better than that of the left.(4)Portal vein blood flow,blood flow velocity,and portal vein diameter of the right graft are higher than the left.(5)Apoptosis may be the reason for the growth difference of dual grafts. |
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