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Currently liver transplantation is the most effective treatment options for patients with end-stage liver disease, but the lack of donor livers limits the development of liver transplantation. With the increasing usage of marginal donor liver such as fatty donor liver, therefore how to make marginal donor liver play a better role has become a new research hotspot. Although the use of fatty liver as a donor can effectively alleviate the contradiction between recipient demand and donor organ shortage, the risk of early post-transplant graft dysfunction also increases. As donor of fatty liver, the degree of steatoidosis should be moderate or below as far as possible, which significantly reduces postoperative complications and promote the recovery of recipients. Meanwhile, it is essential to monitor the lipid indicators post the surgery, and drug intervention can be added if necessary, which may improve the function of the graft.
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Objective To establish a stable model of reduced-size liver transplantations with rejection in rats.Methods Inbred adult male Lewis rats (weighing 220-250 g,8-10 weeks old) served as donors in liver transplantation,and inbred adult male Brown Norway (BN) rats (weighing 220-250 g,8-10 weeks old) served as recipients.Orthotopic group,50％ reduced-size liver transplantation rejection group (50％ group) and 30％ reduced-size liver transplantation rejection group (30％ group)were developed.The 1-,3-,7-and 14-survival rate was observed.Besides,the levels of ALT,AST and TBIL in the peripheral blood plasma were determined.The liver tissues were obtained,and HE staining was used to examine the pathological changes and rejection severity of the liver specimens.Results No significant difference in the operation-related indicators was found among the three groups.The 1-,3-,7-and 14-day survival rate in orthotopic group was as follows:91.20％,86.50％,81.10％ and 75.70％;85.00％,80.00％,62.50％ and 45.00％;65.00％,50.00％,35.00％ and 17.50％,respectively.The 1-,3-,7-and 14-survival rate in 30％ group was significantly lower than in other groups (P＜0.05),and showed no statistical difference between 50％ group and orthotopic group (P＞0.05).The ALT,AST and TBIL levels were significantly higher in 30％ group than in control group at all time points (P＜0.05),but there was no statistical difference between 50％ group and orthotopic group (P ＞ 0.05).All groups were the models of liver transplantations with rejection in rats,and rejection activity index score was increased with time delay.Conclusion Lewis→BN rats are chosen to establish liver transplant rejection model,and by comparing the survival rate and pathological changes among the groups,finally 50％ group is selected to be the model for a follow-up study.
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Objective To study the repairing effect of bone marrow mesenchymal stem cells (BMMSCs) on the donor liver after cardiac death (DCD) under normal temperature mechanical perfusion (NMP) in rats.Methods BMMSCs of Wistar rats were cultured in vitro,and 45-min warm ischemia after cardiac death model was established.The 30 Wistar rats were randomly divided into NMP,NMP + BMMSCs (N + B),cold storage (CS) groups,and the parameters were detected at 2 h and 4 h (n =10).Results N + B group was superior to NMP group and CS group in repairing the liver function and liver pathology including ultrastructure,improving the perfusate acidic environment,and increasing adenosine triphosphate level (P ＜ 0.05).The oxygen consumption of NMP group and N + B group were significantly different after 2h [2 h:(24.35 ±0.64) ml/min vs.(29.33 ±0.47) ml/min;3 h:(25.33 ±0.86) ml/min vs.(30.34 ± 0.49) ml/min;4 h:(26.88 ± 1.07) ml/min vs.(31.76 ± 0.96) ml/min;P ＜ 0.05],suggesting that the liver condition in N + B group was significantly better than that in the other two groups.Conclusion Bone marrow mesenchymal stem cells could obviously repair the DCD grafts under normal temperature mechanical perfusion.
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Objective To investigate the in vitro effects of bone marrow mesenchymal stem cells (BM MSCs) on the replication of HBV with the participation of lymphocytes and to analyze the possible mechanism.Methods The HBV genomic DNA transfected HepG2.2.15 cell line was used to evaluate the HBV replication.Bone marrow and spleen samples were collected from BN rats for the isolation of BM MSCs and T lymphocyte cells, respectively.Five groups of co-culturing with different cells were designed in this study.The cellular activities of lymphocytes and HepG2.2.15 cells were detected at the time of 24 h, 48 h and 72 h after co-culturing by using MTT method.The levels of HBV DNA and HBV cccDNA were detected by real-time polymerase chain reaction ( PCR) .T cell subsets in co-culture were measured by using fluores-cence labeled antibodies and flow cytometry analysis.ELISA was used to detect the levels of cytokines in the supernatant of cultured cells.Results Compared with HepG2.2.15 cells group, BM MSCs+HepG2.2.15 cells and splenic lymphocytes+HepG2.2.15 cells co-culture groups, the levels of HBV DNA and HBV cccDNA were significantly decreased in splenic lymphocytes+BM MSCs+HepG2.2.15 cells co-culture group after 48 h of culture [ HBV DNA: ( 181.000 ±14.731 ) IU/ml vs ( 6270.000 ±300.450 ) IU/ml, (2564.000±231.058) IU/ml, (2433.300±302.379) IU/ml;HBV cccDNA: (4.330×105 ±0.464×105 ) IU/ml vs (11.100×105±0.375×105) IU/ml, (8.930×105±0.778×105) IU/ml, (9.850×105±0.810× 105) IU/ml;P<0.01].The secretion of IFN-γin the supernatant of co-cultured cells was negatively corre-lated with HBV DNA level, but the levels of IL-10 and IL-22 were positively correlated with HBV DNA.The ratio of CD4+/CD8+cells was increased in splenic lymphocytes+BM MSCs+HepG2.2.15 cells co-culture group.The percentage of CD8+cells showed a positive correlation with HBV DNA.Conclusion BM MSCs could inhibit the expression of HBV DNA to enhance the clearance of HBV strains.It might be possibly due to rebalancing of Tc1/Tc2 cells and regulating the expression of autocrine agents and cytokines.