A Study of Serum HSP in Heart Transplanted Rats / 대한이식학회지

PURPOSE: This study was performed to investigate the availability of the serum HSP72 and HSP27 as serologic markers of cardiac allograft rejection through rat heterotopic heart transplatation model. METHODS: Inbred Lewis rats were randomly divided into three groups: the allograft heart transplant group, the isograft heart transplant group, and the sham-operated group. Six animals were studied in each group. In allograft heart tranplant group, the Brown Norway rats were used as donors and in isograft heart tranplant group, the Lewis rats were used as donors. The sera of the allograft heart transplanted rats, isograft heart transplanted rats, and sham- operated rats were collected at preoperative time, 3 days after operation and 6 days after operation, and analyzed for HSP72 and HSP27 by Western blots. Quantifications of band densities were carried out by laser densitometer and the results were expressed as % preoperative densities. RESULTS: The levels of serum HSP72 of 3 days and 6 days after heart transplantation significantly increased in the allograft heart transplant group than in the isograft heart transplant group, respectively (160.2+/-44.8% vs. 109.0+/-34.7%, 276.0+/-72.1% vs. 175.0+/-44.2%, P<0.05). The levels of seum HSP27 of 3 days and 6 days after heart transplantation significantly increased in the allograft heart transplant group than in the isograft heart transplant group, respectively (162.3+/-62.7% vs. 118.4+/-37.0%, 235.7+/-67.1% vs. 127.9+/-40.8%, P<0.05). CONCLUSION: It is concluded that serum HSP72 and HSP27 are useful markers to detect the cardiac allograft rejection.

Donor Cell Repopulation after Bone Marrow Transplantation and Heterotopic Partial Liver Transplantation in Rats Prework for Chimerism

PURPOSE: Improvements in the prevention or control of rejection of kidneys and livers have been largely interchangeable and then applicable. However, the mechanism by which antirejection treatment permits any of these grafts to be accepted has been an immunological enigma. Recently, the exchange of migratory leukocytes between the transplant and the recipient, with consequent long-term cellular chimerism in both has been the basis for acceptance of all whole-organ allografts and xenografts. METHODS: The donors of liver transplants were male Lewis rats weighing 100-150 g in all experiments groups. Male Brown Norway rats were the experimental group and female Lewis were the control group. Heterotopic partial liver transplantation was performed by Lee's method without arterial reconstruction. All procedures were performed under ether anesthesia. Bone marrow was taken from the tibias and femurs and was processed in RPMI 1640. The cell counts of suspensions were 2.5x10(8) per experiment. Genomic DNA prepared from peripheral blood and various tissues. Male Lewis Sry-specific oligonulceotide primers were used. RESULTS: In allogenic liver transplantation with bone marrow transplants (LEW-BN), donor cells were detected in the liver, and the spleen by day 7. However, in rejection cases, donor cell were not detected in any tissues. In isografted transplants (LEW-LEW), after bone marrow transplantation, donor cells were found in lymph nodes, the liver, and peripheral blood. In isografted transplants (LEW-LEW), after liver transplantation donor, cells were only found in the grafts. CONCLUSION: In allogenic liver transplantation with bone marrow transplantation, chimerism induction was augmentedwith bone marrow-derived stem cells. Therefore, it is necessary to have many samples to investigate more precisely chimerism and rejection after liver transplantation with bone marrow transplantation.

Testicular Transplantation in the Rat Model: The Comparison of Testosterone, Gonadotropin, and Histological Change in Allogenic and Isogenic Transplantation Group, and Implantation Group / 대한비뇨기과학회지

PURPOSE: The fate of avascular implants was compared with that of testicular transplants using vascular anastomosis in castrated inbred rats. MATERIALS AND METHODS: Mature inbred male rats were bilaterally castrated and received one testicle as a transplant to the scrotum. The blood supply of the grafts was reestablished by performing an end-to-side anastomosis between an aorto-caval segment of the donor bearing the testicular vessels and the abdominal aorta plus vena cava of the recipient. The average ischemia was 90 min. Lutenizing hormone(LH), follicle-stimulating hormone(FSH) and testosterone were determined by radioimmunoassay in serum samples. RESULTS: In 12 transplant recipients(6 isografts and 6 allografts), a good blood perfusion of the graft was achieved. Out of these, 6 isogenic transplants maintained normal testosterone,LH and FSH during follow up. Implant recipients,however, all displayed a strking early rise of the serum gonadotropins and concomitant castrsted levels of serum testosterone. CONCLUSIONS: Vascular transplantation rather than simple implantation is the method of choice for further research. And in allogenic testicular transplantation, there remains a problem of rejection after the operation, as in any other organ transplantation.

Tacrolimus(FK-506) Nephrotoxicity and the Effect of alpha-Tocopherol in the Rat Renal Isograft / 대한이식학회지

There have been many reports about oxygen free radical injury as a pathogenetic mechanism of CyA nephrotoxicity, but few reports have investigated the relationship between Tacrolimus(FK-506) nephrotoxicity and oxygen free radical injury. Therefore, we decided to evaluate the relationship between Tacrolimus nephrotoxicity and oxygen free radicals, to examine the protective effect of alpha-tocopherol as an antioxidant, and finally to determine the histological changes of these injuries. En bloc resection of the left kidney, left renal artery including a portion of aorta, left renal vein with vena cava, and left ureter including a portion of bladder from male Lewis rats was done, and then preserved in UW solution and stored in the refrigerator at 4oC for 24 hours. After right nephrectomy in the recipients, the harvested organs were transplanted into the right peritoneal cavity using end-to-side anastomoses of the aorta and inferior vena cava between the recipient and donor under a microscope. Also, end-to-end anastomosis of the partly-resected bladders was made between the recipient and donor. After transplantations, rats were divided into 4 groups(I~IV). 2 mg of Tacrolimus per kilogram body weight was injected intramuscularly daily into groups II and III. alpha-Tocopherol was injected intraperitoneally daily in the amount of 20 mg/kg from 2 days prior to transplantation in groups III and IV. The control group(I) received the same amount of saline. 5 or 6 rats from each group were sacrificed at 3 days, 7 days, and 14 days after transplantation, respectively. The grafted and native kidneys were removed for histopathologic examination and the measurement of malondialdehyde(MDA) using a modified TBA method(Ohkawa). Both morphologic renal tubular injury and the increase of MDA due to cold ischemia-reperfusion were highest at 3 days after transplantation, then were alleviated after 7 days. The inhibitory effect of alpha-tocopherol to renal tubular damage from cold ischemia-reperfusion began to appear after 1 week, and was distinct 2 weeks after transplantation. The degree of renal tubular damage was the most severe in Tacrolimus nephrotoxicity, and the frequency of tubulointerstitial nephritis increased with the passage of time, as compared with the ischemia-reperfusion injury(group I). With alpha-tocopherol treatment, ischemia-reperfusion injury as well as Tacrolimus nephrotoxicity was decreased or healed 2 weeks after transplantation, and the amount of MDA was markedly decreased after 1 week. In summary, Tacrolimus nephrotoxicity prolonged the duration of acute tubular necrosis and caused tubulointerstitial nephritis in the rat renal isograft model, which may be the result of aggravation of ischemia-reperfusion injury. That the renal damage due to cold ischemia-reperfusion and Tacrolimus administration was reduced by alpha- tocopherol, indicates that oxidative injury is a pathogenetic mechanism of Tacrolimus nephrotoxicity in this model.

Tacrolimus(FK-506) Nephrotoxicity and the Effect of alpha-Tocopherol in the Rat Renal Isograft / 대한이식학회지

There have been many reports about oxygen free radical injury as a pathogenetic mechanism of CyA nephrotoxicity, but few reports have investigated the relationship between Tacrolimus(FK-506) nephrotoxicity and oxygen free radical injury. Therefore, we decided to evaluate the relationship between Tacrolimus nephrotoxicity and oxygen free radicals, to examine the protective effect of alpha-tocopherol as an antioxidant, and finally to determine the histological changes of these injuries. En bloc resection of the left kidney, left renal artery including a portion of aorta, left renal vein with vena cava, and left ureter including a portion of bladder from male Lewis rats was done, and then preserved in UW solution and stored in the refrigerator at 4oC for 24 hours. After right nephrectomy in the recipients, the harvested organs were transplanted into the right peritoneal cavity using end-to-side anastomoses of the aorta and inferior vena cava between the recipient and donor under a microscope. Also, end-to-end anastomosis of the partly-resected bladders was made between the recipient and donor. After transplantations, rats were divided into 4 groups(I~IV). 2 mg of Tacrolimus per kilogram body weight was injected intramuscularly daily into groups II and III. alpha-Tocopherol was injected intraperitoneally daily in the amount of 20 mg/kg from 2 days prior to transplantation in groups III and IV. The control group(I) received the same amount of saline. 5 or 6 rats from each group were sacrificed at 3 days, 7 days, and 14 days after transplantation, respectively. The grafted and native kidneys were removed for histopathologic examination and the measurement of malondialdehyde(MDA) using a modified TBA method(Ohkawa). Both morphologic renal tubular injury and the increase of MDA due to cold ischemia-reperfusion were highest at 3 days after transplantation, then were alleviated after 7 days. The inhibitory effect of alpha-tocopherol to renal tubular damage from cold ischemia-reperfusion began to appear after 1 week, and was distinct 2 weeks after transplantation. The degree of renal tubular damage was the most severe in Tacrolimus nephrotoxicity, and the frequency of tubulointerstitial nephritis increased with the passage of time, as compared with the ischemia-reperfusion injury(group I). With alpha-tocopherol treatment, ischemia-reperfusion injury as well as Tacrolimus nephrotoxicity was decreased or healed 2 weeks after transplantation, and the amount of MDA was markedly decreased after 1 week. In summary, Tacrolimus nephrotoxicity prolonged the duration of acute tubular necrosis and caused tubulointerstitial nephritis in the rat renal isograft model, which may be the result of aggravation of ischemia-reperfusion injury. That the renal damage due to cold ischemia-reperfusion and Tacrolimus administration was reduced by alpha- tocopherol, indicates that oxidative injury is a pathogenetic mechanism of Tacrolimus nephrotoxicity in this model.

Renal Ischemia/Reperfusion Injury and Cyclosporine Nephrotoxicity in the Rat Renal Isograft Model: A Pilot Study / 대한이식학회지

Renal allografts frequently suffer from ischemia/reperfusion injury, which is a major cause of delayed graft function in renal transplantation(Tx). Cyclosporine(CsA) is known to aggravate ischemic injury, which may further heighten graft dysfunction. To know the histopathologic features of renal ischemic/reperfusion injury and cyclosporoine nephrotoxicity, we performed renal Tx between Lewis rats with cold ischemia and with/without CsA. Rats were sacrificed 3, 5, 7 days, 2, 3 and 4 weeks post-Tx. Control rats received sham operation. The kidney was processed for light microscopy and stained with H-E, PAS. Furthermore, to know the distribution of thioredoxin peroxidase(TPx), a recently cloned antioxidant in this model, the kidney tissue was stained with antibodies against three subtypes of TPx; NKEF-A /PAG(TPx A), NKEF-B/TPx(TPx B) and Mer 5. Renal isografts showed acute tubular necrosis from 3 days and recovery by 7 days, which was prolonged in CsA treated rats with signs of tubular and vascular toxicity. In sham operated rats, TPx A was distributed in all tubular segments, most prominently in distal tubules(DT). TPx B was stained in DT and collecting ducts(CD) exclusively. Mer 5 was present mainly in S3 segment. Glomerular or vascular expression was not found. In isografts TPx A expression was increased in both proximal(PT) and DT, markedly in the nonnecrotic S1 segment till 1 week postTx and returned to normal pattern by 2 weeks. TPx B and Mer 5 expression were increased till 5 days postTx with stronger staining in DT than in PT. CsA sustained the tubular expression of TPx till 4 weeks postTx. In summary, TPx expression was increased in renal tubules of rat renal isografts suffering cold ischemia, and more prolonged with CsA therapy. Marked increase of TPx A expression in S1 segment of ischemic kidneys may indicate resistance against oxidant injury, especially in S1 segment.