Objective To observe the dynamic histopathologic changes of acute rejection in rat orthotopic liver transplantation (OLT) model after tacrolimus discontinued and provide some prediction and evaluation data for clinical acute rejection after liver transplantation. Methods Kamada two-cuff technique was used to establish 60 rat OLT model， and male DA rats, male Lewis rats were used as donors and recipients respectively. Therapeutic amount of tacrolimus (0.05 mg/kg, twice per day, continued for 8 d, 1 d before operation and 7 d after operation, intragastric administrated) was administrated to recipients, then continuously half dose was decreased every day beginning from day 8 after operation and tacrolimus administration was stopped on day 13. Liver tissues were collected on day 7, 14, 21, and 28 after liver transplantation. Histopathologic changes and rejection activity index (RAI) of liver tissues were observed, survival time of recipients was calculated. Results Owing to protection effects of tacrolimus, liver tissues displayed no significant histopathologic changes of acute rejection in 7 d after OLT, while typical acute rejection histopathologic changes began to be observed on day 14 after OLT due to tacrolimus discontinuation. On day 14, 21, and 28, RAI were 3.7±0.9, 6.3±0.9, and 8.1±0.7 respectively. Survival time of recipients was (20.85±0.71) d with a median of 21 d. Conclusion Acute rejection could be induced in rat OLT model after tacrolimus discontinuation, and data collected from this model shows some extent of predictive value and assessment value for clinical liver acute rejection.
Abstract: Objective To study the antiacute rejection effect of Pachymic acid (PA) in heart transplantation rats, in order to select a new antirejection medicine with low side effect from traditional Chinese medicine. Methods We established the model by transplanting Wistar rats (32,donor) heart allografts into the abdomen of SD rats (32,receptor). The homologous hearttransplanted rats were then randomly divided into 4 groups with 16 rats in each group. Olive oil solution with PA 1 mg/(kg·d), PA 10 mg/(kg·d), Cyclosporine (CsA) 5 mg/(kg·d) and olive oil solution 0.5 ml/(kg·d) were respectively given intragastrically to lowdosage PA group, highdosage PA group, CsA group and the control group till the end of observation. Survival time of heart allografts, heart beating and the histological changes of allografts were examined and serum level of interleukin2 (IL-2) and interferon-γ (IFN-γ) were determined by enzymelinked immunosorbent assay (ELISA). Results Survival time in the highdosage PA group, the lowdosage PA group and the CsA group were 24.90±0.99 d, 15.50±1.60 d and 26.80±0.88 d respectively, which is much better than the control group (6.10±1.10 d, q=22.363, P=0.000; q=44.793, P=0.000; q=49.272,P=0.000). IL-2 serum level in the highdosage PA group, the lowdosage PA group and the CsA group were all lower than that in the control group (q=14.483, P=0.000; q=3.705, P=0.000; =21.418,P=0.000), whileIL-2 serum level in the highdosage group was lower than that in the lowdosage group (q=10.778,P=0.000). Similarly, IFN-γ serum level in the first three groups were all lower than that in the control group (q=16.508,P=0000; q=4.281, P=0.000;q=19.621, P=0.000) and IFNγ serum level in the highdosage group was also lower than that in the lowdosage group (q=14.975, P=0.000). Pathological examination 7 days after the surgery showed that pathologic lesion was much more relieved in the two PA groups and the CsA group than the control group. Conclusion Acute rejection of heart transplantation can be effectively suppressed by PA.
Abstract:Objective To investigate immunoinh.ibitory effects of paclitaxel on acute rejection of allogeneic heart transplantation in rats. Methods Heterotopic abdominal cardiac transplantation was performed from Wistar rats to SD rats. Seventy recipients were randomly divided into five groups,14 rats in each group. Control group： rats didn＇t receive any immunoinhibitory drug; group Ⅰ ： low-dose paclitaxel （0.75 mg/kg · d） was injected intraperitoneally; group Ⅱ ： high-dose paclitaxel （1.5 mg/kg ·d） was injected intraperitoneally; group Ⅲ ： cyclosporin A（CsA, 5 mg/ kg·d） was administered orally; group Ⅳ ： low-dose paclitaxel （0. 75 mg/kg · d） was injected intraperitoneally in combination with CsA （5 mg/kg · d administered orally）. General conditions of recipient, allograft survival and pathologic lesion at 7th day posttransplantation were observed. Results Allograft survival in treating groups were significantly prolonged compared with control group （P〈 0. 05）. Moreover, allograft survival in group IV was significantly prolonged compared with those in group Ⅰ and group Ⅲ （P〈0.05）. On 7th day posttransplantation, cardiac allograft looked swollen and International Society for Heart and Lung Transplantation （ISHLT） score was 3 or 4 in control group; cardiac allograft beat vigorously, showed pink in color and felt tender in group Ⅰ and group Ⅱ , ISHLT-score was 2 or 3. Compared to control group, pathologic lesion of grafts in group Ⅰ and group Ⅱ were significantly relieved （P〈0.05）. Cardiac allograft beat well and ISHLT-score was 2 in group Ⅲ. Cardiac allograft looked as normal and beat vigorously, ISHLT-score was less than 2 in group IV ; the protective effects on cardiac allograft was better than those in group Ⅰ and group Ⅱ （P〈0. 05）. Conclusion Paclitaxel could obviously suppress acute rejection and prolong survival of rat cardiac allograft. Paclitaxel and CsA has synergistic effect on prevention acute rejection.
Objective To study the effect of recombinant lentiviral vector mediated human hepatocyte growth factor (hHGF) gene-modified bone marrow mesenchymal stem cells (BMSCs) on the immunological rejection after allograft l iver transplantation in rats, and to reveal the mechanism of immune tolerance. Methods Eight male Sprague Dawley (SD)rats of clean grade (aged 3 to 4 weeks, weighing 75-85 g) were selected for the isolation and culture of BMSCs; 64 adult male SD rats of clean grade (weighing 200-250 g) were used as donors; and 64 adult male Wistar rats of clean grade (weighing 230-280 g) were used as receptors. After establ ishing a stable model of rat allogeneic l iver transplantation, 1 mL sal ine, 2 ×106/mL of BMSCs 1 mL, 2 × 106/mL of BMSCs/green fluorescent protein 1 mL, and 2 × 106/mL of BMSCs/hHGF 1 mL were injected via the portal vein in groups A, B, C, and D respectively. Then the survival time of the rats was observed. The hepatic function was determined and the histological observation of the l iver was performed. The hHGF mRNA expression was detected by RT-PCR, the level of cytokine including hHGF, interleukin 2 (IL-2), IL-4, IL-10, and interferon γ (IFN-γ) by ELISA assay, the level of apoptosis by TUNEL method, and the expression level of prol iferating cell nuclear antigen (PCNA) by immunohistochemical method. Results The survival time of group D was significantly higher than that of groups A, B, and C (P lt; 0.01); the survival time of groups B and C was significantly higher than that of group A (P lt; 0.01), but there was no significant difference between group B and group C (P gt; 0.05). RT-PCR demonstrated the transcription of hHGF mRNA in the grafts of group D; the serum cytokine hHGF reached to (6.2 ± 1.0) ng/mL. Compared with groups B and C, group D exhibited significant inhibitory effect, significantly improved l iver function, and showed mild acute rejection. In addition, the levels of cytokine IL-2 and IFN-γ decreased; the levels of cytokine IL-4 and IL-10 increased; the level of apoptosis reduced; and the expression level of PCNA increased. Except for the expression of IL-4 (P gt; 0.05), there were significant differences in the other indexes between group D and groups B, C (P lt; 0.05). Conclusion BMSCs/hHGF implanting to rat l iver allograft via portal vein can induce immune tolerance. Compared with injection of BMSCs alone, BMSCs/hHGF treatment can alleviate acute rejection and prolong the survival time significantly. The immunosuppressive effect of BMSCs/hHGF is correlated with Th2 shifts up of Th1/Th2 shift, reduced apoptosis, promoted l iver regeneration.
Objective To establish the rat orthotopic liver transplantation model by characterizing the blood supply of hepatic artery with the Cuff skill and the modified arterial sleeve anastomosis, to explore the possible mechanisms of acute rejection and the express of Fractalkine (Fkn) in the early stage after hepatic allograft operation. Methods SD rats were selected as donors and Wistar rats as receptor for the rejection model of orthotopic liver transplantation. Recipient rats were divided into 2 groups randomly after operationand the drugs were given intraperitoneally once a day in each group. In the experimental group, cyclosporine A (CsA) was delivered with 3 mg/kg. In the control group, only normal saline was given with 3 ml/kg. Condition of survivals were observed. The rejection actvity index (RAI) and the expression of Fkn of liver tissue were observed after 3rd, 5th and 7th days in 5 rats. The rest of rats in each group were fed and given drug or normal saline until they were died and the mean survival time were recorded. Results There were 18 survivals in control group, and 19 in experimental group after liver transplantation. Condition of survivals in experimental group was better than that of control group. The mean survival times of experimental group(19.50±4.51 days) was significantly longer than that of control group(7.60±1.60 days), showing statistically significant difference (P＜0.05). After 3rd, 5th and 7th days of transplantation, RAI of control group were 3.80±0.35,5.90±0.87 and 7.50±1.30,respectively;RAI of experimental group were 3.10±0.21,3.90±0.41 and 4.50±0.52.Therewasstatistically significant difference in RAI between 2 groups on the 7th day after transplantation (Plt;0.01). On the 3rd,5th and 7th days after transplantation, the Fkn of control group was 8.20±0.57,21.30±3.30 and 25.70±4.91, and that of experimental group was 8.30±0.56,10.30±0.67 and 11.70±1.23. There were statistically significant differences in Fkn between 2 groups on the 5th, 7th days after transplantation (Plt;0.01). Conclusion Fkn is a participant inacute rejection after the rat orthotopic liver transplantation and can be chosen as a useful target in the diagnosis of acute rejection. CsA has immunosuppressive property in the condition of acute rejection in the rat orthotopic liver transplantation, which may be result from the decreased the level of Fkn.
Objective To summarize the current advance of xenotransplantation. Methods Relevant literatures about current advance of xenotransplantation published recently domestic and abroad were collected and reviewed. Results Major progress of xenotransplantation had been made in the understanding of xenoimmunobiology in the last two decades and in the threshold of clinical application. However, many problems of immunological rejection were still needed to be explored and resolved. Conclusion Xenotransplantation as a transplantation source has an extensive potential to resolve the shortage of transplanted organs for end-stage organ failure, how to suppress rejection and prolong survival of grafts more effectively is a focal point of search in the future.
【Abstract】Objective To explore the effect and indication of splenectomy in liver transplantation. Methods From January 2001 to April 2006, 260 patients underwent piggyback orthotopic liver transplantation (PBOLT), and 28 patients had undergone combined PBOLT and splenectomy (splenectomy group). These patients were compared to 56 randomly selected non-splenectomy patients from the same transplant period, meaningly two controls were selected for every non-splenectomy case. Two groups were analyzed with respect to rate of infection and survival rate, as well as biopsy-proven acute allograft rejection within 30 days after transplantation. Results Rate of infection in the splenectomy group was higher than that in the non-splenectomy patients (85.7% vs 55.4%, P＜0.05). Acute rejection and survival rates in the splenectomy group were lower than those in the non-splenectomy patients (3.6% vs 14.3%, P＜0.05; 46.4% vs 82.1%, P＜0.05). Conclusion Concomitant splenectomy with PBOLT has a significantly higher patient mortality rate; it is mainly due to its septic complications. At present, unless there is a certain indication for splenectomy, this procedure is not recommended.
Objective To discuss the differentiation between transient intrahepatic cholestasis (TIHC) and acute rejection (AR) after liver transplantation． Methods Characteristics and the changes (before and within 21 d after transplantation) of alanine aminotransferase (ALT) and direct bilirubin (DBIL) in 30 patients undergone liver transplantation were observed. These patients were divided into TIHC group and AR group following the diagnosis criteria, and the serum levels of ALT and DBIL were compared respectively on day 1 before liver transplantation, day 3, 7 and 21 after liver transplantation． Results Compared with day 3 after transplantation in the TIHC group， DBIL significantly ascended while ALT was changeless on day 7 after transplantation. But in the AR group， DBIL ascended significantly and ALT showed an increasing tendency on day 7 after transplantation． After appropriate therapy, DBIL and ALT of two groups both descended significantly on day 21 after transplantation． Conclusion The changes of DBIL and ALT are available for the differentiation between TIHC and AR after transplantation.
Objective To observe the effects of Thymosin α1 （Tα1） on acute rejection after liver transplantation and immune function of T cells. Methods Twenty recipients of liver transplantation due to primary hepatic carcinoma were divided into two groups: Tα1 group (n=10) and control group (n=10). Tα1 group received subcutaneous injection of Tα1 1.6 mg on the first day after liver transplantation and then twice a week for at least one month. Both Tα1 group and control group took same immunodepressants. Core biopsies were carried to compare the incidence rate of acute rejection between Tα1 group and control group. Peripheral T cellular immune function in these two groups was detected on 1 d before, 1 week, 2 weeks and 1 month after transplantation. Results There was not significant difference of incidence rate of acute rejection between Tα1 group and control group (Pgt;0.05). In the Tα1 group, CD4+, CD8+ lymphocyte cell counts and the CD4+/CD8+ ratio were significantly higher than those in the control group in 2 weeks and 1 month after transplantation (P＜0.05). Conclusion Use of Tα1 in recipients who also takes rountine immunosuppressants dose not increase the risk of occurring acute rejection after liver transplantation. Tα1 can significantly increase CD4+, CD8+ counts and CD4+/CD8+ ratio, which shows that Tα1 may improve recipients’ cellular immune function.
Objective To observe the expression of heat shock protein 70 (HSP70) in human liver after hepatic transplantation, and to study its correlation with the occurrence and progression of acute allograft rejection.Methods Fifteen biopsy specimen of allograft liver after transplantation were collected and divided into three groups according to their pathological changes: control group (no rejection), mild acute rejection group, and moderate/serious acute rejection group. The expressions of HSP70 in grafts were detected by using immunohistochemical method and imaging analysis. Results HSP70 was expressed in all 3 groups, and appeared mainly in hepatocellular cytoplasm. The immunohistochemical imaging analysis of HSP70 showed: integral optical density (IOD) which was 30.99±11.14 in the control group was lower than that in the mild acute rejection group (68.84±21.37) and that in the moderate/serious acute rejection group (71.82±19.99), P＜0.01； and the IOD in the moderate/serious acute rejection group was higher than that in the mild acute rejection group (P＜0.05). Conclusion HSP70 plays a role in cellular protection for allograft liver, and the continuously increasing expression of HSP70 in graft maybe closely relates to the occurrence and progression of acute allograft rejection.